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Zhang Z, Wu O, Ying J, Jin Y, Wang H, Tian H, Chen Q, Chen L, Tao C, Lou C, Jones M, Wang X, Makvandi P, Shen S, Li B, Wu A. Regulation of diabetic disc degeneration: The role of AGEAT/miR-204-5p/Mapk4 axis in nucleus pulposus cells' mitochondrial function and apoptosis. Cell Signal 2025; 133:111857. [PMID: 40381974 DOI: 10.1016/j.cellsig.2025.111857] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2025] [Revised: 04/22/2025] [Accepted: 05/07/2025] [Indexed: 05/20/2025]
Abstract
Chronic low back pain associated with intervertebral disc degeneration (IVDD) is significantly aggravated in patients with diabetes mellitus (DM); however, the underlying molecular mechanisms remain unclear. This study explored the role of the long non-coding RNA AGEAT (AGE-associated transcript) in the pathogenesis of DM-associated IVDD. Whole-transcriptome sequencing of rat nucleus pulposus cells (NPCs) treated with advanced glycation end products (AGEs) revealed a time-dependent upregulation of AGEAT. AGEAT overexpression induced NPC apoptosis, mitochondrial dysfunction, and extracellular matrix (ECM) degradation. Mechanistically, RNA fluorescence in situ hybridization localized AGEAT to the cytoplasm, where it acted as a competing endogenous RNA (ceRNA) by directly binding miR-204-5p, thereby relieving repression of its target Mapk4. Silencing AGEAT via siRNA significantly reduced apoptosis, restored mitochondrial function, and preserved ECM integrity. In vivo, intra-discal injection of AAV-sh-AGEAT in diabetic IVDD rats significantly improved disc integrity, as evidenced by a reduction in MRI Pfirrmann grade and histological preservation of NPC density and collagen II content. Collectively, these findings establish AGEAT as a key ceRNA that exacerbates diabetic IVDD via the miR-204-5p/Mapk4 axis, promoting NPC apoptosis, mitochondrial dysfunction, and ECM degradation. Targeting this pathway-through AGEAT silencing or miR-204-5p activation-represents a promising therapeutic strategy for mitigating diabetes-associated disc degeneration. This study reveals the critical role of the AGEAT/miR-204-5p/Mapk4 axis in the progression of DM-associated IVDD, suggesting a potential therapeutic strategy for its treatment.
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Affiliation(s)
- Zhiguang Zhang
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Department of Orthopedics Medicine Center, Jinhua Municipal Central Hospital, Zhejiang, China
| | - Ouqiang Wu
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Jiahao Ying
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Yuxin Jin
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Hui Wang
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Department of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou, China
| | - Haijun Tian
- Department of Orthopaedic Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, China
| | - Qizhu Chen
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Linjie Chen
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Chen Tao
- Department of Orthopaedics, Key Laboratory of Spine and Spinal Cord, Injury Repair and Regeneration of Ministry of Education Tongji Hospital, Tongji University School of Medicine, School of Life Science and Technology, Tongji University, Shanghai 200065, China
| | - Chao Lou
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Morgan Jones
- Spine Unit, The Royal Orthopaedic Hospital, Bristol Road South, Northfield, Birmingham B31 2AP, United Kingdom
| | - Xiangyang Wang
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China
| | - Pooyan Makvandi
- The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, 324000 Quzhou, Zhejiang, China; Institute for Bioengineering, School of Engineering, The University of Edinburgh, Edinburgh, United Kingdom
| | - Shuying Shen
- Department of Orthopaedic Surgery, Sir Run Run Shaw Hospital, Zhejiang University school of medicine & KeyLaboratory of Musculoskeletal System Degeneration and Regeneration Translational Research of Zhejiang, Hangzhou 310016, China
| | - Bin Li
- Orthopedic Institute, Department of Orthopaedic Surgery, The First Affiliated Hospital, School of Biology & Basic Medical Sciences, Suzhou Medical College, Soochow University, Suzhou, Jiangsu 215006, China
| | - Aimin Wu
- Department of Orthopaedics, Key Laboratory of Structural Malformations in Children of Zhejiang Province, Key Laboratory of Orthopaedics of Zhejiang Province, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
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Zhao Y, Chen L, Jiang S, Wu Z, Xiang Q, Lin J, Tian S, Sun Z, Sun C, Li W. Exosomes derived from MSCs exposed to hypoxic and inflammatory environments slow intervertebral disc degeneration by alleviating the senescence of nucleus pulposus cells through epigenetic modifications. Bioact Mater 2025; 49:515-530. [PMID: 40206196 PMCID: PMC11979484 DOI: 10.1016/j.bioactmat.2025.02.046] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2024] [Revised: 02/11/2025] [Accepted: 02/28/2025] [Indexed: 04/11/2025] Open
Abstract
Intervertebral disc degeneration (IDD) is the leading cause of low back pain, which places heavy burdens on society and individuals. Surgical intervention is the conventional therapy for IDD, but patients who undergo surgery face relatively high risks of recurrence and complications. Therefore, a relatively less invasive and efficient treatment for IDD is urgently needed. In this study, we constructed a novel nanobiomaterial, named Hi-Exos, to slow IDD. Hi-Exos are exosomes derived from mesenchymal stem cells exposed to hypoxic and inflammatory environments. Hi-Exos could relieve the senescence of nucleus pulposus cells and slow IDD through an epigenetic modification mechanism by introducing the epigenetic factor miR-221-3p into senescent nucleus pulposus cells to reduce DDIT4 expression and inhibit the activation of NF-κB signalling pathway. This study provided a novel strategy for IDD treatment involving the use of Hi-Exos to deliver miR-221-3p to reduce the senescence of nucleus pulposus cells and repair IDD via epigenetic modifications.
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Affiliation(s)
- Yongzhao Zhao
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Longting Chen
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Shuai Jiang
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Zhenquan Wu
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Qian Xiang
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Jialiang Lin
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Shuo Tian
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Zhuoran Sun
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Chuiguo Sun
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
| | - Weishi Li
- Department of Orthopaedics, Peking University Third Hospital, Beijing, China
- Beijing Key Laboratory of Spinal Disease Research, Beijing, China
- Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, China
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Hu H, Xie T. Gastrodin Alleviates Lumbar Intervertebral Disc Degeneration by Suppressing the NF-κB and MAPK Pathways. Cell Biochem Biophys 2025; 83:2009-2021. [PMID: 39821825 DOI: 10.1007/s12013-024-01612-2] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 11/01/2024] [Indexed: 01/19/2025]
Abstract
Intervertebral disc degeneration (IDD) is the main pathological factor resulting in low back pain (LBP), the leading cause of disability globally. Inflammatory response and extracellular matrix (ECM) degradation are critical pathological features in the development of IDD. Gastrodin (GAS), a phenol compound isolated from Gastrodia elata Blume, plays an anti-inflammatory role in experimental models of multiple human diseases. Our study aimed to elucidate whether GAS alleviates TNF-α-induced inflammation in nucleus pulposus (NP) cells and IDD in vivo. The cytotoxicity of GAS was assessed by CCK-8 assay. Rat primary NP cells were stimulated with TNF-α to induce inflammatory response. The expression of proinflammatory cytokines, catabolic genes, and anabolic genes was detected by RT-qPCR, western blotting, and immunofluorescence staining. NF-κB and MAPK pathway activation was determined through western blotting and immunofluorescence staining. The IDD rat model was established by using percutaneous needle puncture. The therapeutic effects of GAS were confirmed by histology analysis. We found that TNF-α stimulation enhanced proinflammatory cytokine (COX2, iNOS, IL-6, and IL-1β) expression in NP cells, which was reversed by GAS treatment. GAS offset TNF-α-induced upregulation in catabolic gene (MMP3, MMP9, and MMP13) expression and downregulation in anabolic gene (Collagen II, SOX9, and Aggrecan) expression. The loss of ECM in TNF-α-treated NP cells was mitigated by GAS treatment. Mechanically, GAS abolished TNF-α-induced increase in p-IKKα, p-IKKβ, p-IκBα, p-p65, p-ERK, p-p38, and p-JNK protein levels in NP cells. In puncture-induced IDD rat models, GAS administration improved intervertebral disc (IVD) structure, increased Collagen II expression, and reduced the levels of proinflammatory factors in IVDs. Overall, GAS alleviates the inflammation and ECM degradation in NP cells via inhibiting NF-κB and MAPK pathway activation and alleviates IDD in vivo, which may be a novel treatment strategy for IDD.
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Affiliation(s)
- Huimin Hu
- Department of Orthopedic, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, 430014, Hubei Province, China
| | - Tian Xie
- Department of Orthopedic, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, 430014, Hubei Province, China.
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Guo W, Mu K, Geng JC, Xing HY, Dong Y, Liu WD, Wang SC, Shi JX, Xing BR, Zhao JY, Li XM. ATF1 and miR-27b-3p drive intervertebral disc degeneration through the PPARG/NF-κB signaling axis. Commun Biol 2025; 8:751. [PMID: 40369110 PMCID: PMC12078598 DOI: 10.1038/s42003-025-08186-6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2024] [Accepted: 05/07/2025] [Indexed: 05/16/2025] Open
Abstract
Intervertebral disc degeneration (IDD) is a primary cause of degenerative disc disease; however, the mechanisms underlying it remain unknown. Although great efforts have been made to develop new regenerative therapies, their clinical success is limited. Recent research has indicated that microRNAs (miRNAs) are significantly involved in the progression of IDD. Investigating the role of miRNA intervention in IDD could facilitate the development of therapeutic strategies based on miRNAs. However, circulating miRNAs have not yet been recognized as standard biomarkers for IDD. In this study, we observed that the expression of miR-27b-3p was elevated in the blood and nucleus pulposus (NP) tissue of patients with IDD. Furthermore, reducing the expression of miR-27b-3p was shown to impede the progression of IDD. MiR-27b-3p could reduce the expression of collagen II and ACAN and promote the expression of MMP13 and ADAMT-5 in vitro and in vivo. miR-27b-3p aggravated IDD progression by directly targeting peroxisome proliferator-activated receptor gamma (PPARG), a negative regulator of the NF-κB signal pathway. This study also established that PPARG serves a protective role in IDD. The overexpression of PPARG was able to mitigate the detrimental effects caused by miR-27b-3p in NP cells and animal models of IDD, indicating that miR-27b-3p facilitates the progression of IDD through its interaction with PPARG. Additionally, the transcription factor ATF1 was found to enhance the expression of miR-27b-3p by targeting its promoter region, thereby promoting the degenerative impact of miR-27b-3p on NP cells. Given that miR-27b-3p can promote IDD both in vitro and in vivo, it holds potential as a biomarker, and the inhibition of miR-27b-3p expression may represent a novel therapeutic target for IDD.
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Affiliation(s)
- Wei Guo
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China.
- Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research, Cangzhou, PR China.
- Hebei Province Integrated Traditional Chinese and Western Medicine 3D Printing Technology Innovation Center, Cangzhou, PR China.
| | - Kun Mu
- Department of Breast Surgery, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
| | - Jing-Chao Geng
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
| | - Hai-Yang Xing
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
| | - Yu Dong
- Department of Anaesthesiology, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
| | - Wen-Dong Liu
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
- Hebei Province Integrated Traditional Chinese and Western Medicine 3D Printing Technology Innovation Center, Cangzhou, PR China
| | - Shuan-Chi Wang
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
- Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research, Cangzhou, PR China
| | - Jia-Xiao Shi
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
- Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research, Cangzhou, PR China
| | - Bao-Rui Xing
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
- Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research, Cangzhou, PR China
- Hebei Province Integrated Traditional Chinese and Western Medicine 3D Printing Technology Innovation Center, Cangzhou, PR China
| | - Jian-Yong Zhao
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China
- Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research, Cangzhou, PR China
- Hebei Province Integrated Traditional Chinese and Western Medicine 3D Printing Technology Innovation Center, Cangzhou, PR China
| | - Xiao-Ming Li
- Department of Orthopaedics, Hebei Province Cangzhou Hospital of Integrated Traditional Chinese Medicine-Western Medicine, Cangzhou, PR China.
- Hebei Key Laboratory of Integrated Traditional and Western Medicine in Osteoarthrosis Research, Cangzhou, PR China.
- Hebei Province Integrated Traditional Chinese and Western Medicine 3D Printing Technology Innovation Center, Cangzhou, PR China.
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Lin L, Li D, Cai G, Zheng G, Huang D, Liu H, Lin S, Zhao F. Exploring the molecular mechanisms underlying intervertebral disc degeneration by analysing multiple datasets. Sci Rep 2025; 15:14748. [PMID: 40289127 PMCID: PMC12034803 DOI: 10.1038/s41598-025-98070-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/03/2024] [Accepted: 04/09/2025] [Indexed: 04/30/2025] Open
Abstract
The purpose of this study was to explore the genetic characteristics and immune cell infiltration related to intervertebral disc degeneration through multidataset analysis, predict potential therapeutic drugs, and provide a theoretical basis for clinical treatment. The gene expression profile data of the GSE70362, GSE186542, and GSE245147 datasets were downloaded from the Gene Expression Omnibus (GEO) database, and the hub genes were identified through differentially expressed gene analysis, Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) functional annotation and Mendelian randomization analysis were performed. Hub genes and immune cells were identified. Infiltration status was determined through GSEA and GSVA to clarify the specific signalling pathways associated with key genes and explore the potential molecular mechanisms by which key genes affect disease progression. The key genes were reversely predicted using miRNA grid construction and transcription factor regulation, and genes related to disease regulation were obtained from the GeneCards database. Finally, the differentially expressed genes were used for drug prediction through the Connectivity Map database to identify potential drugs for the treatment of intervertebral disc degeneration. The feasibility of the predicted drugs was tested by molecular docking technology. Real-time quantitative PCR was used to confirm the expression of key genes in the tissue samples.A total of 126 differentially expressed genes were identified in the GEO database, and 4 differentially expressed hub genes (COL6A2, DCXR, GLRX, and PDGFRB) were identified through bioinformatics methods. Immune infiltration analysis revealed that NK cells, macrophages, and eosinophils were activated during IVDD, whereas mast cells and T cells were suppressed. GO and KEGG analyses revealed that key genes are involved in the development of this disease through signalling pathways such as the glycolysis pathway, the oxidative phosphorylation pathway, the cholesterol regulatory pathway, and the haem metabolism pathway. Analysis of the constructed miRNA grid revealed that key genes are jointly regulated by multiple transcription factors, among which the most important motif is cisbp_M5578. Disease regulation-related genes were obtained through the GeneCards database, analysis of the correlation with key genes was performed, and the expression levels of the two mRNA and miRNA were significantly correlated. Finally, drug prediction performed through the Connectivity Map database revealed that drugs such as Abt-751, LY-2183240, podophyllotoxin, and vindesine can alleviate or even reverse the disease state. Finally, we collected 10 IVDD and 10 healthy disc tissue samples, and the RT‒qPCR results were consistent with the bioinformatics results. We identified COL6A2, DCXR, GLRX, and PDGFRB as key genes involved in IVDD. In addition, drugs such as Abt-751 are expected to control and reverse the progression of the disease. In the future, these key genes and predicted drugs may provide new directions for further mechanistic studies as well as new therapies for IVDD patients.
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Affiliation(s)
- Longquan Lin
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China.
| | - Da Li
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China
| | - Gangfeng Cai
- Department of Neurosurgery, Fujian Medical University Union Hospital, Fujian, 350000, China.
| | - Gengyang Zheng
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China
| | - Dianfeng Huang
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China
| | - Hua Liu
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China
| | - Shunxin Lin
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China
| | - Feng Zhao
- Department of Orthopaedics, The 910th Hospital of PLA, Quanzhou, 362000, China
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Han L, Li F, Wu H, Wang W, Chen P, Xia W, Liu Y, Sun K, Lin W. Targeting FABP4 to Inhibit AGEs-RAGE/NF-κB Signalling Effectively Ameliorates Nucleus Pulposus Dysfunction and Angiogenesis in Obesity-Related Intervertebral Disc Degeneration. Cell Prolif 2025:e70021. [PMID: 40090836 DOI: 10.1111/cpr.70021] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2024] [Revised: 02/16/2025] [Accepted: 02/28/2025] [Indexed: 03/18/2025] Open
Abstract
Intervertebral disc degeneration (IVDD) is a primary contributor to low back pain, posing significant social and economic burdens. Increasing evidence shows that obesity contributes to IVDD, yet the underlying mechanisms remain elusive. Here, we firstly revealed a causal correlation between obesity and IVDD via a two-sample mendelian randomization analysis and identified fatty acid-binding protein 4 (FABP4) as the potential regulator to associate IVDD and obesity. Elevated FABP4 expression promoted extracellular matrix (ECM) disequilibrium and angiogenesis to exacerbate IVDD progression. Genetically knocking out or pharmacologically inhibiting FABP4 in high-fat diet-induced mice alleviated IVDD. Mechanistically, obesity activated the mammalian target of rapamycin complex 1 (mTORC1), which upregulated FABP4 expression, leading to the accumulation of advanced glycation end-products (AGEs) in intervertebral disc tissue. AGEs further activated the NF-κB signalling pathway, exacerbating ECM degradation and neovascularization. Conversely, rapamycin-mediated inhibition of mTORC1 suppressed FABP4 expression in nucleus pulposus cells (NPCs), alleviating IVDD in vivo. Collectively, our findings reveal a critical role of the obesity-induced mTORC1-FABP4 axis in ECM degradation and angiogenesis during IVDD progression. Targeting FABP4 may represent a promising therapeutic strategy for IVDD in obese individuals.
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Affiliation(s)
- Lin Han
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
- Department of Orthopedic Surgery, Third Affiliated Hospital of Naval Medical University, Shanghai, China
| | - Fudong Li
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Huiqiao Wu
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Weiheng Wang
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Peiwen Chen
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Weicheng Xia
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Yang Liu
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Kaiqiang Sun
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
| | - Wenbo Lin
- Department of Orthopedic Surgery, Shanghai Changzheng Hospital, Naval Medical University, Shanghai, China
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Luo J, Jin G, Cui S, Wang H, Liu Q. Regulating macrophage phenotypes with IL4I1-mimetic nanoparticles in IDD treatment. J Nanobiotechnology 2025; 23:175. [PMID: 40050923 PMCID: PMC11884037 DOI: 10.1186/s12951-025-03241-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2024] [Accepted: 02/18/2025] [Indexed: 03/10/2025] Open
Abstract
Intervertebral disc degeneration (IDD) is a degenerative spinal condition characterized by disc structural damage, narrowing of joint spaces, and nerve root compression, significantly reducing patients' quality of life. To address this challenge, a novel therapeutic strategy was developed using cellulose supramolecular hydrogel as a carrier to deliver IL4I1-modified MΦ membrane biomimetic nanoparticles (CHG@IL4I1-MNPs) to target tissues. This hydrogel exhibits excellent biocompatibility and mechanical properties while enabling sustained drug release in the degenerative disc microenvironment, enhancing therapeutic outcomes. CHG@IL4I1-MNPs effectively regulate MΦ polarization by promoting M2 MΦ activation, thereby improving immune microenvironment balance. Animal studies demonstrated that CHG@IL4I1-MNPs alleviated symptoms of IDD, reduced inflammation, and supported tissue repair, highlighting its potential to reduce reliance on long-term medication and improve quality of life. The strategy uniquely combines nanoparticle technology with immunomodulation, achieving precise targeting of MΦs. Beyond IDD, this approach offers potential applications in other immune-related diseases, providing a versatile platform for nanomedicine. This study introduces an innovative method to treat IDD and advances the integration of immunotherapy and nanotechnology, offering both clinical benefits and new directions for future research. These findings hold strong potential for improving patient outcomes and expanding treatment options for related diseases.
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Affiliation(s)
- Jiaying Luo
- School of Life Sciences and Biopharmaceuticals, Shenyang Pharmaceutical University, Shenyang, 110016, China
| | - Guoxin Jin
- Department of Orthopedics, Shengjing Hospital of China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, Liaoning Province, 110000, China
| | - Shaoqian Cui
- Department of Orthopedics, Shengjing Hospital of China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, Liaoning Province, 110000, China
| | - Huan Wang
- Department of Orthopedics, Shengjing Hospital of China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, Liaoning Province, 110000, China
| | - Qi Liu
- Department of Orthopedics, Shengjing Hospital of China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, Liaoning Province, 110000, China.
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Shi WH, Zou HS, Wang XY, Lu J, Yu HQ, Zhang PP, Huang LL, Chu PC, Liang DC, Zhang YN, Li B. Identification and Experimental Verification of Potential Immune Cell-Associated Gene Biomarkers in Human Intervertebral Disc Degeneration. J Pain Res 2025; 18:993-1007. [PMID: 40034105 PMCID: PMC11873021 DOI: 10.2147/jpr.s505859] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2024] [Accepted: 01/31/2025] [Indexed: 03/05/2025] Open
Abstract
Objective Intervertebral disc degeneration (IDD) is one of the most common diseases in the elderly population. Recently, immune disorders have been considered to play an important role in IDD. This study aimed to conduct a bioinformatic analysis to identify biomarkers associated with IDD immune cells. Methods We obtained the gene expression profiles of IDD by downloading the Gene Expression Omnibus Series (GSE)150408 and GSE124272 from the Gene Expression Omnibus (GEO) database. IDD and immune cell-related hub genes were identified via multiple bioinformatics analyses, and their diagnostic performance was evaluated using receiver operating characteristic (ROC) analysis. In addition, a long non-coding RNA (lncRNA)-signature gene co-expression network was constructed. Finally, the diagnostic accuracy of the biomarkers was verified using Real-time quantitative PCR (RT-qPCR). Results ASAP1-IT1, IKZF2, KLHL14, lnc-C10orf131-1, and LOC101927805 were identified as signature genes of IDD. Further, ROC analysis revealed that the five signature gene models had a strong ability to discriminate between the IDD and healthy control samples. We also found that the five signature genes were significantly associated with immune-inflammatory feedback, cell cycle, and skeletal system. Furthermore, an lncRNA signature gene network was constructed to reveal the regulatory mechanisms of the biomarkers. Finally, RT-qPCR results verified that IKZF2 and KLHL14 were significantly downregulated in patients with IDD, and ASAP1-IT1 was significantly upregulated. Conclusion This study identified ASAP1-IT1, IKZF2, and KLHL14 as the key signature genes of IDD. These key hub genes may serve as potential therapeutic targets for IDD.
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Affiliation(s)
- Wei-Han Shi
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Hui-Shuang Zou
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Xiang-Yu Wang
- Department of Pain Medicine, First Medical Center, PLA General Hospital, Beijing, 100000, People’s Republic of China
| | - Jie Lu
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Hua-Qi Yu
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Ping-Ping Zhang
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Li-Li Huang
- Department of Scientific Research Management, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Peng-Cheng Chu
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Da-Chuan Liang
- Department of Scientific Research Management, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Ya-Ning Zhang
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
| | - Bin Li
- Department of Orthopedics, Shanxi Medical University Seventh Affiliated Hospital, Linfen People’s Hospital, Linfen, Shanxi, 041000, People’s Republic of China
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9
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Yan X, Ding JY, Zhang RJ, Wang YX, Zhou LP, Zhang HQ, Kang L, Jia CY, Liu XY, Shen CL. FSTL1 accelerates nucleus pulposus-derived mesenchymal stem cell apoptosis in intervertebral disc degeneration by activating TGF-β-mediated Smad2/3 phosphorylation. J Transl Med 2025; 23:232. [PMID: 40011941 PMCID: PMC11863476 DOI: 10.1186/s12967-025-06231-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2024] [Accepted: 02/11/2025] [Indexed: 02/28/2025] Open
Abstract
BACKGROUND Intervertebral disc degeneration (IVDD) is the leading cause of low back pain, and repair using nucleus pulposus-derived mesenchymal stem cells (NP-MSCs) represents a promising therapeutic approach. However, both endogenous and transplanted NP-MSCs demonstrate limited proliferative capacity, increased apoptosis, and reduced resilience to the harsh microenvironment within the degenerative intervertebral disc (IVD). METHODS RNA sequencing (RNA-seq) was utilized to identify genes and associated mechanisms that mediate the responses of NP-MSCs to acidic conditions. Western blotting, qPCR, and immunofluorescence were used to evaluate follistatin-like 1 (FSTL1) expression in NP-MSCs. Apoptosis and extracellular matrix (ECM) anabolism were assessed via flow cytometry, TUNEL staining and Western blotting, while the TGF-β/Smad2/3 pathway was analyzed using Western blotting and immunofluorescence. FSTL1 knockdown with small interfering RNA (siRNA) was performed to determine its role in apoptosis and ECM regulation. The FSTL1 siRNA pretreatment was assessed in a puncture-induced rat IVDD model using MRI and histological staining. RESULTS Using RNA-seq, we identified FSTL1 as the primary acid-responsive gene in NP-MSCs. We further observed elevated FSTL1 expression in NP-MSCs isolated from degenerative IVDs in both humans and rats compared to normal IVDs. Acidic conditions upregulated FSTL1 expression in NP-MSCs in a pH-dependent manner. Notably, recombinant FSTL1 was shown to enhance cellular apoptosis and disrupt ECM metabolism. Conversely, silencing FSTL1 with siRNA reduced NP-MSC apoptosis and improved ECM anabolism. Importantly, TGF-β pathway inhibition partially reversed the pro-apoptotic and ECM catabolism effects of FSTL1. In the rat model of IVDD, pretreatment of NP-MSCs with FSTL1 siRNA significantly suppressed IVDD progression. CONCLUSIONS This study provides novel insights into the mechanistic role of FSTL1 in acid-induced apoptosis of NP-MSCs and its contribution to the progression of IVDD. These findings offer valuable perspectives for developing targeted therapeutic strategies to mitigate IVDD progression.
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Affiliation(s)
- Xu Yan
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Jing-Yu Ding
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Ren-Jie Zhang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Yan-Xin Wang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Lu-Ping Zhou
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Hua-Qing Zhang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Liang Kang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Chong-Yu Jia
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China
| | - Xiao-Ying Liu
- School of Life Sciences, Anhui Medical University, 81 Meishan Road, Hefei, Anhui, 230032, China.
| | - Cai-Liang Shen
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China.
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, Anhui, 230022, China.
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, China.
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10
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Zhang F, Cui D, Wang Z, Li Y, Wang K, Lu H, Yu H, Jiao W, Cui X. NOX4 Regulates NLRP3 by Inhibiting the Ubiquitination of LRRC8A to Promote Ferroptosis in Nucleus Pulposus Cells. Inflammation 2025:10.1007/s10753-025-02253-0. [PMID: 39909992 DOI: 10.1007/s10753-025-02253-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/13/2024] [Revised: 12/20/2024] [Accepted: 01/23/2025] [Indexed: 02/07/2025]
Abstract
Intervertebral disc degeneration (IDD) is a significant contributor to low back pain, imposing a considerable socioeconomic burden. Ferroptosis, a novel form of cell death driven by iron and characterized by the accumulation of reactive oxygen species (ROS), has been associated with the progression of IDD. Nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) has been widely recognized as a pivotal factor promoting ferroptosis across various diseases; however, its precise role in the pathogenesis of IDD remains incompletely understood. Our experimental findings demonstrated a marked upregulation of NOX4 in degenerated cells, accompanied by elevated ROS levels and a diminished mitochondrial membrane potential, indicating the participation of ferroptosis. Furthermore, the expression of the critical regulatory factor GPX4 was reduced, while ACSL4 levels were significantly increased, further corroborating the involvement of ferroptosis. Functional loss and gain experiments revealed that NOX4 overexpression augmented ferroptosis and ROS production while promoting the secretion of inflammatory cytokines. Subsequent studies indicated that the knockdown of NOX4 could reverse tert-butyl hydroperoxide (TBHP)-induced ferroptosis. Mass spectrometry analysis identified leucine-rich repeat-containing 8A (LRRC8A) as an interacting protein of NOX4, and further validation confirmed that they co-regulate Nod-like receptor pyrin domain-3 (NLRP3) activation through their interaction. Utilizing a rat model of intervertebral disc degeneration, we further corroborated the role of NOX4 in IDD. This study provides theoretical support for the potential application of NOX4-targeting drugs in the treatment of IDD.
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Affiliation(s)
- Feng Zhang
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
| | - Di Cui
- Medical School of Fuyang, Normal University, No. 100, Qinghe West Road, Yingzhou District, Fuyang, 236000, Anhui, China
| | - Zhaodong Wang
- Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical University, No.2600, Donghai Dadao, Bengbu, 233000, Anhui, China
- Department of Orthopedics, the First Affiliated Hospital of Bengbu Medical University, No. 287,Changhuai Road, Bengbu, 233000, Anhui, China
| | - Yifei Li
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
| | - Kangkang Wang
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
| | - Haitao Lu
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China
| | - Haiyang Yu
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
| | - Wei Jiao
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
| | - Xilong Cui
- Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
- Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
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11
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Cheng Z, Gan W, Xiang Q, Zhao K, Gao H, Chen Y, Shi P, Zhang A, Li G, Song Y, Feng X, Yang C, Zhang Y. Impaired degradation of PLCG1 by chaperone-mediated autophagy promotes cellular senescence and intervertebral disc degeneration. Autophagy 2025; 21:352-373. [PMID: 39212196 PMCID: PMC11759519 DOI: 10.1080/15548627.2024.2395797] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2023] [Revised: 08/18/2024] [Accepted: 08/20/2024] [Indexed: 09/04/2024] Open
Abstract
Defects in chaperone-mediated autophagy (CMA) are associated with cellular senescence, but the mechanism remains poorly understood. Here, we found that CMA inhibition induced cellular senescence in a calcium-dependent manner and identified its role in TNF-induced senescence of nucleus pulposus cells (NPC) and intervertebral disc degeneration. Based on structural and functional proteomic screens, PLCG1 (phospholipase C gamma 1) was predicted as a potential substrate for CMA deficiency to affect calcium homeostasis. We further confirmed that PLCG1 was a key mediator of CMA in the regulation of intracellular calcium flux. Aberrant accumulation of PLCG1 caused by CMA blockage resulted in calcium overload, thereby inducing NPC senescence. Immunoassays on human specimens showed that reduced LAMP2A, the rate-limiting protein of CMA, or increased PLCG1 was associated with disc senescence, and the TNF-induced disc degeneration in rats was inhibited by overexpression of Lamp2a or knockdown of Plcg1. Because CMA dysregulation, calcium overload, and cellular senescence are common features of disc degeneration and other age-related degenerative diseases, the discovery of actionable molecular targets that can link these perturbations may have therapeutic value.Abbreviation: ATRA: all-trans-retinoic acid; BrdU: bromodeoxyuridine; CDKN1A/p21: cyclin dependent kinase inhibitor 1A; CDKN2A/p16-INK4A: cyclin dependent kinase inhibitor 2A; CMA: chaperone-mediated autophagy; DHI: disc height index; ER: endoplasmic reticulum; IP: immunoprecipitation; IP3: inositol 1,4,5-trisphosphate; ITPR/IP3R: inositol 1,4,5-trisphosphate receptor; IVD: intervertebral disc; IVDD: intervertebral disc degeneration; KD: knockdown; KO: knockout; Leu: leupeptin; MRI: magnetic resonance imaging; MS: mass spectrometry; N/L: NH4Cl and leupeptin; NP: nucleus pulposus; NPC: nucleus pulposus cells; PI: protease inhibitors; PLC: phospholipase C; PLCG1: phospholipase C gamma 1; ROS: reactive oxygen species; RT-qPCR: real-time quantitative reverse transcription PCR; SA-GLB1/β-gal: senescence-associated galactosidase beta 1; SASP: senescence-associated secretory phenotype; STV: starvation; TMT: tandem mass tag; TNF: tumor necrosis factor; TP53: tumor protein p53; UPS: ubiquitin-proteasome system.
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Affiliation(s)
- Zhangrong Cheng
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Weikang Gan
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Qian Xiang
- Department of Orthopedics, Peking University Third Hospital, Beijing, China
| | - Kangcheng Zhao
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Haiyang Gao
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Yuhang Chen
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Pengzhi Shi
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Anran Zhang
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Gaocai Li
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Yu Song
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Xiaobo Feng
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Cao Yang
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
| | - Yukun Zhang
- Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China
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12
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Wei YF, Zhang HL, Li LZ, Lv Y, Li H, Li Z, Yu FL, Jiang T, Zhang TY, Xin F, Ma C, Ren YX. Sirt1 blocks nucleus pulposus and macrophages crosstalk by inhibiting RelA/Lipocalin 2 axis. J Orthop Translat 2025; 50:30-43. [PMID: 39758288 PMCID: PMC11699611 DOI: 10.1016/j.jot.2024.11.008] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/30/2024] [Revised: 09/26/2024] [Accepted: 11/25/2024] [Indexed: 01/07/2025] Open
Abstract
Background Intervertebral disc degeneration (IVDD) stands as a primary pathophysiological driver of low back pain, yet no therapeutic intervention effectively arrests its progression. Evidence shows that certain Sirt1 agonists may confer protective effects on intervertebral discs, but the underlying mechanisms remain unclear. This study aims to delineate the interaction between Sirt1 and the inflammatory microenvironment, offering potential novel avenues for IVDD prevention and treatment. Methods In vitro IL-1β-induced nucleus pulposus cells (NPCs) degenerative model and in vivo a mouse annulus fibrosus needle puncture model in Sirt1 transgenic (Sirt1TG) and the same litter WT mice were used to investigate the role of Sirt1 in homeostasis and inflammation. Mechanistic insights were obtained through RNA sequencing, co-immunoprecipitation (Co-IP), luciferase assays, and chromatin immunoprecipitation-(ChIP)-PCR. A co-culture system of Raw264.7 and NPCs was employed to assess the involvement of Lipocalin 2. Results Our study demonstrated reduced Sirt1 expression in degenerating human nucleus pulposus (NP) tissue. Both in vitro and in vivo data revealed that NP-specific overexpression of Sirt1 inhibited extracellular matrix degradation and inflammation. Mechanistically, Sirt1 suppressed the acetylation of RelA/p65 at lysine 310 and phosphorylation at serine 536, with the C-terminus of Sirt1 and the RHD-NLS domain of RelA mediating to their interaction. Furthermore, NPCs-derived Lipocalin 2 was identified as a cytokine involved in macrophage chemotaxis and M1 polarization to exacerbate inflammation. Conclusion Our work revealed that Sirt1 negatively regulates Lipocalin 2, thereby ameliorating the inflammatory milieu and blocking NPCs and macrophages crosstalk. The Translational Potential of this Article This study illuminates the crucial role and molecular mechanisms of Sirt1 in regulating the NP microenvironment. These insights shed light on strategies for the prevention and treatment of IVDD-related herniation and low back pain. By pinpointing specific biological targets, the screening of smallmolecule compounds with significant clinical implications can be facilitated. This translational innovation promises to optimize cells communication within intervertebral disc microenvironment via localized drug delivery, potentially improving patient outcomes and satisfaction following spinal fusion or discectomy surgeries.
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Affiliation(s)
- Yi-Fan Wei
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
| | - He-Long Zhang
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
| | - Ling-Zhi Li
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
| | - You Lv
- Department of Orthopaedics, Lianyungang Clinical College of Nanjing Medical University, 6 Zhenhua East Rd, Lianyungang, 221000, China
| | - He Li
- Department of Sports Medicine, Lianyungang Clinical College of Nanjing Medical University, 6 Zhenhua East Rd, Lianyungang, 221000, China
| | - Zhi Li
- Department of Orthopaedics, Geriatric Hospital of Nanjing Medical University, 65 Jiangsu Rd, Nanjing, 210024, China
| | - Feng-Lei Yu
- Department of Trauma and Orthopaedics, The First People's Hospital of Kunshan, 566 East Qianjin Rd, Suzhou, 215000, China
| | - Tao Jiang
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
| | - Tian-You Zhang
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
| | - Feng Xin
- Department of Orthopaedics, Xuzhou Cancer Hospital, 131 Huancheng Rd, Xuzhou, 221005, China
| | - Cheng Ma
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
| | - Yong-Xin Ren
- Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing, 210029, China
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13
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Zhou Q, Pu X, Qian Z, Chen H, Wang N, Wang S, Feng Z, Zhu Z, Wang B, Qiu Y, Sun X. Nuclear receptor Rev-erbα alleviates intervertebral disc degeneration by recruiting NCoR-HDAC3 co-repressor and inhibiting NLRP3 inflammasome. Cell Prolif 2024; 57:e13720. [PMID: 39045886 PMCID: PMC11628727 DOI: 10.1111/cpr.13720] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2024] [Revised: 06/27/2024] [Accepted: 07/08/2024] [Indexed: 07/25/2024] Open
Abstract
Intervertebral discs (IVDs) are rhythmic tissues that experience daily low-load recovery. Notably, aging and abnormal mechanical stress predispose IVDs to degeneration due to dysrhythmia-induced disordered metabolism. Meanwhile, Rev-erbα acts as a transcriptional repressor in maintaining biorhythms and homeostasis; however, its function in IVD homeostasis and degeneration remains unclear. This study assessed the relationship between low Rev-erbα expression levels and IVD degeneration. Rev-erbα deficiency accelerated needle puncture or aging-induced IVD degeneration, characterized by increased extracellular matrix (ECM) catabolism and nucleus pulposus (NP) cell apoptosis. Mechanistically, Rev-erbα knockdown in NP cells aggravated rhIL1β-induced NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activation, exacerbating the imbalanced ECM and NP cell apoptosis. Meanwhile, blocking NLRP3 inflammasome activation mitigated Rev-erbα deficiency and needle puncture-induced IVD degeneration. Particularly, Rev-erbα mediated the transcriptional repression of the NLRP3 inflammasome via the ligand heme-binding of nuclear receptor co-repressor (NCoR) and histone deacetylase 3 (HDAC3) complex. Thus, the increased expression of Rev-erbα in NP cells following short-term rhIL1β treatment failed to inhibit NLRP3 transcription in vitro owing to heme depletion. Pharmacological activation of Rev-erbα in vivo and in vitro alleviated IVD degeneration by altering the NLRP3 inflammasome. Taken together, targeting Rev-erbα may be a potential therapeutic strategy for alleviating IVD degeneration and its related diseases.
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Affiliation(s)
- Qingshuang Zhou
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Xiaojiang Pu
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Zhuang Qian
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Haojie Chen
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Nannan Wang
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Sinian Wang
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Zhenhua Feng
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Zezhang Zhu
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Bin Wang
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Yong Qiu
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
| | - Xu Sun
- Division of Spine Surgery, Department of Orthopedic SurgeryNanjing Drum Tower Hospital Clinical College of Jiangsu UniversityNanjingChina
- Division of Spine Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical SchoolNanjing UniversityNanjingChina
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Ní Néill T, Barcellona MN, Wilson N, O'Brien FJ, Dixon JE, Curtin CM, Buckley CT. In vitro and ex vivo screening of microRNA combinations with enhanced cell penetrating peptides to stimulate intervertebral disc regeneration. JOR Spine 2024; 7:e1366. [PMID: 39726900 PMCID: PMC11669629 DOI: 10.1002/jsp2.1366] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/08/2023] [Revised: 02/23/2024] [Accepted: 03/23/2024] [Indexed: 12/28/2024] Open
Abstract
Background Low back pain (LBP) is predominantly caused by degeneration of the intervertebral disc (IVD) and central nucleus pulposus (NP) region. Conservative treatments fail to restore disc function, motivating the exploration of nucleic acid therapies, such as the use of microRNAs (miRNAs). miRNAs have the potential to modulate expression of discogenic factors, while silencing the catabolic cascade associated with degeneration. To deliver these miRNAs, nonviral cell penetrating peptides (CPPs) are gaining favor given their low immunogenicity and strong targeting ability. Single miRNA therapies have been investigated for IVD repair, however dual miRNA delivery strategies have not been commonly examined and may augment regeneration. Materials and methods Transfection of four pro-discogenic miRNAs (miRNA mimics:140-5p; 149-5p and inhibitors: 141-3p; 221-3p) and dual delivery of six miRNA pairings was performed using two CPPs, RALA and GET peptide (FLR), in primary rat NP monolayer culture, and in an ex vivo organ culture model of rat caudal discs. Protein expression of discogenic (aggrecan, collagen type II, and SOX9) and catabolic markers (ADAMTS5 and MMP13) were assessed. Results Monolayer investigations signified enhanced discogenic marker expression following dual miRNA delivery, signifying a synergistic effect when compared to single miRNA transfection. Utilization of an appropriate model was emphasized in our ex vivo organ culture experiment, revealing the establishment of a regenerative microenvironment characterized by reduced catabolic enzyme activity and enhanced matrix deposition, particularly following concurrent delivery of FLR-miRNA-149-5p mimic and miRNA-221-3p inhibitor. Bioinformatics analysis of miRNA-149-5p mimic and miRNA-221-3p inhibitor identified distinct targets, pathways, and interactions, suggesting a mode of action for this amplified response. Conclusion Our findings suggest the potential of FLR-miRNA-149-5p + miRNA-221-3p inhibitor to create an anti-catabolic niche within the disc to foster regeneration in moderate cases of disc degeneration, which could be utilized in further studies with the overarching aim of developing treatments for LBP.
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Affiliation(s)
- Tara Ní Néill
- Trinity Centre for Biomedical EngineeringTrinity Biomedical Sciences Institute, Trinity College Dublin, The University of DublinDublinIreland
- Discipline of Mechanical, Manufacturing and Biomedical EngineeringSchool of Engineering, Trinity College Dublin, The University of DublinDublinIreland
- Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland & Trinity College Dublin, The University of DublinDublinIreland
| | - Marcos N. Barcellona
- Trinity Centre for Biomedical EngineeringTrinity Biomedical Sciences Institute, Trinity College Dublin, The University of DublinDublinIreland
- Discipline of Mechanical, Manufacturing and Biomedical EngineeringSchool of Engineering, Trinity College Dublin, The University of DublinDublinIreland
- Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland & Trinity College Dublin, The University of DublinDublinIreland
| | - Niamh Wilson
- Trinity Centre for Biomedical EngineeringTrinity Biomedical Sciences Institute, Trinity College Dublin, The University of DublinDublinIreland
- Discipline of Mechanical, Manufacturing and Biomedical EngineeringSchool of Engineering, Trinity College Dublin, The University of DublinDublinIreland
- Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland & Trinity College Dublin, The University of DublinDublinIreland
| | - Fergal J. O'Brien
- Trinity Centre for Biomedical EngineeringTrinity Biomedical Sciences Institute, Trinity College Dublin, The University of DublinDublinIreland
- Discipline of Mechanical, Manufacturing and Biomedical EngineeringSchool of Engineering, Trinity College Dublin, The University of DublinDublinIreland
- Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland & Trinity College Dublin, The University of DublinDublinIreland
- Tissue Engineering Research Group, Department of Anatomy and Regenerative MedicineRCSIDublinIreland
| | - James E. Dixon
- Regenerative Medicine and Cellular TherapiesThe University of Nottingham Biodiscovery Institute (BDI), School of Pharmacy, University of NottinghamNottinghamUK
- NIHR Nottingham Biomedical Research CentreUniversity of NottinghamNottinghamUK
| | - Caroline M. Curtin
- Trinity Centre for Biomedical EngineeringTrinity Biomedical Sciences Institute, Trinity College Dublin, The University of DublinDublinIreland
- Discipline of Mechanical, Manufacturing and Biomedical EngineeringSchool of Engineering, Trinity College Dublin, The University of DublinDublinIreland
- Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland & Trinity College Dublin, The University of DublinDublinIreland
- Tissue Engineering Research Group, Department of Anatomy and Regenerative MedicineRCSIDublinIreland
| | - Conor T. Buckley
- Trinity Centre for Biomedical EngineeringTrinity Biomedical Sciences Institute, Trinity College Dublin, The University of DublinDublinIreland
- Discipline of Mechanical, Manufacturing and Biomedical EngineeringSchool of Engineering, Trinity College Dublin, The University of DublinDublinIreland
- Advanced Materials and Bioengineering Research (AMBER) CentreRoyal College of Surgeons in Ireland & Trinity College Dublin, The University of DublinDublinIreland
- Tissue Engineering Research Group, Department of Anatomy and Regenerative MedicineRCSIDublinIreland
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15
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Zhang Q, Li Z, Zhou S, Li J. Regenerative Outcomes of Combining siCOL1A2 Hydrogel with Acupuncture in a Rat Model of Chronic Intervertebral Disc Degeneration. Bioengineering (Basel) 2024; 11:1066. [PMID: 39593726 PMCID: PMC11591507 DOI: 10.3390/bioengineering11111066] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/25/2024] [Revised: 09/04/2024] [Accepted: 09/25/2024] [Indexed: 11/28/2024] Open
Abstract
Intervertebral disc degeneration (IVDD) is a significant cause of chronic pain and disability, necessitating innovative therapeutic strategies. This study investigates the combined effect of a novel siCOL1A2-encapsulated hydrogel and acupuncture on IVDD in a rat model. We developed a hydrogel system, siCOL1A2-encapsulated G5-PBA hydrogel (siCOL1A2@G5-PBA@Gel), designed for sustained siRNA delivery to the degenerated discs and assessed its therapeutic efficacy alongside acupuncture treatment. Key inflammatory genes were identified through RNA-seq analysis, with COL1A2 highlighted as a crucial regulator of inflammatory responses in IVDD. Our in vivo experiments involved treating rats with hydrogel alone, acupuncture alone, and combining both. The treatments were evaluated through behavioral pain assessments, imaging techniques (X-ray and MRI), and histological analyses. Results indicated that the combination therapy significantly alleviated pain, reduced inflammation, and promoted disc regeneration more effectively than individual treatments. The hydrogel proved biocompatible and facilitated targeted gene silencing, while acupuncture enhanced therapeutic outcomes by improving local blood circulation and modulating inflammatory responses. These findings suggest that integrating siCOL1A2 hydrogel with acupuncture offers a promising approach to treating IVDD.
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Affiliation(s)
- Qianfu Zhang
- Department of Integrated Traditional and Western Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China; (Q.Z.); (S.Z.)
| | - Zhixuan Li
- Technical Management Department, IIJ Global Solutions China Inc., Shanghai 200031, China;
| | - Sihan Zhou
- Department of Integrated Traditional and Western Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China; (Q.Z.); (S.Z.)
| | - Ji Li
- Department of Integrated Traditional and Western Medicine, Huashan Hospital, Fudan University, Shanghai 200040, China; (Q.Z.); (S.Z.)
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16
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Zhao Y, Xia Q, Zhu L, Xia J, Xiang S, Mao Q, Dong H, Weng Z, Liao W, Xin Z. Mapping knowledge structure and themes trends of non-surgical treatment in intervertebral disc degeneration. Heliyon 2024; 10:e36509. [PMID: 39286189 PMCID: PMC11402762 DOI: 10.1016/j.heliyon.2024.e36509] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2024] [Revised: 03/23/2024] [Accepted: 08/16/2024] [Indexed: 09/19/2024] Open
Abstract
Background Intervertebral disc degeneration (IDD) is a chronic disabling disease caused by degeneration of nucleus pulposus cells, decreased activity and the number of nucleus pulposus cells, decreased extracellular matrix, and infiltration of inflammatory factors, resulting in low back and leg pain. Recent studies have shown that non-surgical treatment is of great significance in reversing the progression of degenerative disc disease, and there are more relevant literature reports. However, there is no bibliometric analysis in this area. This study aimed to describe the knowledge structure and thematic trends of non-surgical treatment methods for IDD through bibliometrics. Methods Articles and reviews on non-surgical treatment of disc degeneration from 1998 to 2022 were collected on the Web of Science. VOSviewer 1.6.18, CiteSpace 6.1.R3, R package "bibliometrix" and two online analysis platforms were used for bibliometric and visual literature analysis. Results 961 articles were screened for inclusion, including 821 articles and 140 reviews. The analysis of our study shows that publications in the non-surgical treatment of disc degeneration are increasing annually, with publications coming mainly from North America and Asia, with China and the United States dominating. Huazhong Univ Sci & Technol and Wang K are the most prolific institutions and authors, respectively, and Le Maitre CL is the most co-cited author. However, there is less collaboration between institutions in different countries. Spine is both the most published and the most cited journal. According to the co-citation and co-occurrence analysis results, "mesenchymal stem cells," "exosomes," "medication," and "tissue engineering" are the current research hotspots in this field. Conclusions This study employs bibliometric analysis to explore the knowledge structure and trends of non-surgical treatments for IDD from 2013 to 2022. Key research hotspots include mesenchymal stem cells, exosomes, medication, and tissue engineering. The number of publications, especially from China and the USA, has increased significantly, though international collaboration needs improvement. Influential contributors include Wang K and the journal Spine. These findings provide a comprehensive overview and highlight important future directions for the field.
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Affiliation(s)
- Yan Zhao
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Qiuqiu Xia
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Lu Zhu
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Jiyue Xia
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Shaojie Xiang
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Qiming Mao
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Huaize Dong
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Zijing Weng
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Wenbo Liao
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
| | - Zhijun Xin
- Department of Orthopedic Surgery, Affiliated Hospital of Zunyi Medical University, Zunyi, 563000, Guizhou, China
- Institut Curie, PSL Research University, CNRS UMR3244, Dynamics of Genetic Information, Sorbonne Université, 75005, Paris, France
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Chen X, Chen K, Hu J, Dong Y, Zheng M, Hu Q, Zhang W. Multiomics analysis reveals the potential of LPCAT1-PC axis as a therapeutic target for human intervertebral disc degeneration. Int J Biol Macromol 2024; 276:133779. [PMID: 38992527 DOI: 10.1016/j.ijbiomac.2024.133779] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/22/2024] [Revised: 06/25/2024] [Accepted: 07/08/2024] [Indexed: 07/13/2024]
Abstract
Intervertebral disc degeneration (IDD) is a highly prevalent musculoskeletal disorder that is associated with considerable morbidity. However, there is currently no drug available that has a definitive therapeutic effect on IDD. In this study, we aimed to identify the molecular features and potential therapeutic targets of IDD through a comprehensive multiomics profiling approach. By integrating transcriptomics, proteomics, and ultrastructural analyses, we discovered dysfunctions in various organelles, including mitochondria, the endoplasmic reticulum, the Golgi apparatus, and lysosomes. Metabolomics analysis revealed a reduction in total phosphatidylcholine (PC) content in IDD. Through integration of multiple omics techniques with disease phenotypes, a pivotal pathway regulated by the lysophosphatidylcholine acyltransferase 1 (LPCAT1)-PC axis was identified. LPCAT1 exhibited low expression levels and exhibited a positive correlation with PC content in IDD. Suppression of LPCAT1 resulted in inhibition of PC synthesis in nucleus pulposus cells, leading to a notable increase in nucleus pulposus cell senescence and damage to cellular organelles. Consequently, PC exhibits potential as a therapeutic agent, as it facilitates the repair of the biomembrane system and alleviates senescence in nucleus pulposus cells via reversal of downregulation of the LPCAT1-PC axis.
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Affiliation(s)
- Xi Chen
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China
| | - Kun Chen
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China
| | - Jun Hu
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China
| | - Yijun Dong
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China
| | - Menglong Zheng
- Department of medical imaging, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China
| | - Qingsong Hu
- Department of Hepatobiliary Surgery, Anhui Province Key Laboratory of Hepatopancreatobiliary Surgery, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China.
| | - Wenzhi Zhang
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, China.
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18
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Tu H, Gao Q, Zhou Y, Peng L, Wu D, Zhang D, Yang J. The role of sirtuins in intervertebral disc degeneration: Mechanisms and therapeutic potential. J Cell Physiol 2024; 239:e31328. [PMID: 38922861 DOI: 10.1002/jcp.31328] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2023] [Revised: 04/27/2024] [Accepted: 05/13/2024] [Indexed: 06/28/2024]
Abstract
Intervertebral disc degeneration (IDD) is one of the main causes of low back pain, which affects the patients' quality of life and health and imposes a significant socioeconomic burden. Despite great efforts made by researchers to understand the pathogenesis of IDD, effective strategies for preventing and treating this disease remain very limited. Sirtuins are a highly conserved family of (NAD+)-dependent deacetylases in mammals that are involved in a variety of metabolic processes in vivo. In recent years, sirtuins have attracted much attention owing to their regulatory roles in IDD on physiological activities such as inflammation, apoptosis, autophagy, aging, oxidative stress, and mitochondrial function. At the same time, many studies have explored the therapeutic effects of sirtuins-targeting activators or micro-RNA in IDD. This review summarizes the molecular pathways of sirtuins involved in IDD, and summarizes the therapeutic role of activators or micro-RNA targeting Sirtuins in IDD, as well as the current limitations and challenges, with a view to provide possible solutions for the treatment of IDD.
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Affiliation(s)
- Heng Tu
- State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Qian Gao
- State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Yumeng Zhou
- State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Li Peng
- Key Laboratory of Bio-Resource & Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, Sichuan, China
| | - Dan Wu
- State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Demao Zhang
- Institute of Biomedical Engineering, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, Sichuan, China
| | - Jing Yang
- State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, Department of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
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19
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Yu XJ, Bai XF, Qu YK, Wang SX, Zhang J, Yang W, Wang S, Yang Y, Wang YG, Hao DJ, Zhao YT. Unveiling the Therapeutic Potential of hUCMSC-Derived EVs in Intervertebral Disc Degeneration through MALAT1/ miR-138-5p/SLC7A11 Coexpression Regulation. ACS Biomater Sci Eng 2024; 10:4839-4854. [PMID: 39079050 DOI: 10.1021/acsbiomaterials.3c01944] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 08/13/2024]
Abstract
Intervertebral disc degeneration (IVDD) is a prevalent chronic condition causing spinal pain and functional impairment. This study investigates the role of extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUCMSCs) in regulating IVDD. Using RNA-seq, we analyzed differential expressions of lncRNA and miRNA in nucleus pulposus tissues from various mouse groups. We identified key regulatory molecules, MALAT1 and miRNA-138-5p, which contribute to IVDD. Further experiments demonstrated that MALAT1 can up-regulate SLC7A11 expression by competitively binding to miR-138-5p, forming a MALAT1/miR-138-5p/SLC7A11 coexpression regulatory network. This study elucidates the molecular mechanism by which hUCMSC-derived EVs regulate IVDD and could help develop novel therapeutic strategies for treating this condition. Our findings demonstrate that hUCMSCs-EVs inhibit ferroptosis in nucleus pulposus cells, thereby improving IVDD. These results highlight the therapeutic potential of hUCMSCs-EVs in ameliorating the development of IVDD, offering significant scientific and clinical implications for new treatments.
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Affiliation(s)
- Xiao-Jun Yu
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Xiao-Fan Bai
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Yun-Kun Qu
- Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
| | - Shan-Xi Wang
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Jianwei Zhang
- Department of Orthopedics, the First People's Hospital of Tianshui City, Tianshui 741000, Gansu Province, China
| | - Wenlong Yang
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Sibo Wang
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Yuli Yang
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
| | - Ying-Guang Wang
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Ding-Jun Hao
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
| | - Yuan-Ting Zhao
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an 710054, Shaanxi, China
- Shaanxi Key Laboratory of Spine Bionic Treatment, Xi'an 710054, Shaanxi, China
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20
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Wang D, Lu K, Zou G, Wu D, Cheng Y, Sun Y. Attenuating intervertebral disc degeneration through spermidine-delivery nanoplatform based on polydopamine for persistent regulation of oxidative stress. Int J Biol Macromol 2024; 274:132881. [PMID: 38838900 DOI: 10.1016/j.ijbiomac.2024.132881] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/24/2023] [Revised: 05/20/2024] [Accepted: 06/02/2024] [Indexed: 06/07/2024]
Abstract
As one of the most widespread musculoskeletal diseases worldwide, intervertebral disc degeneration (IVDD) remains an intractable clinical problem. Currently, oxidative stress has been widely considered as a significant risk factor in the IVDD pathological changes, and targeting oxidative stress injury to improve the harsh microenvironment may provide a novel and promising strategy for disc repair. It is evident that spermidine (SPD) has the ability to attenuate oxidative stress across several disease models. However, limited research exists regarding its impact on oxidative stress within the intervertebral disc. Moreover, enhancing the local utilization rate of SPD holds great significance in IVDD management. This study aimed to develop an intelligent biodegradable mesoporous polydopamine (PDA) nanoplatform for sustained release of SPD. The obtained PDA nanoparticles with spherical morphology and mesoporous structure released loaded-therapeutic molecules under low pH and H2O2. Combined treatment with SPD loaded into PDA nanoparticles (SPD/PDA) resulted in better therapeutic potential than those with SPD alone on oxidative stress injury. Furthermore, both SPD and SPD/PDA could induce anti-inflammatory M2 macrophage polarization. Upon injection into degenerative IVDs, the SPD/PDA group achieved a good repair efficacy with a long-term therapeutic effect. These findings indicated that the synergized use of SPD with responsive drug delivery nanocarriers may steadily scavenge reactive oxygen species and provide an effective approach toward the treatment of IVDD.
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Affiliation(s)
- Dongliang Wang
- Department of Orthopedics, Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Department of Orthopedics, Yancheng First People's Hospital of Jiangsu Province, Yancheng, Jiangsu, China
| | - Kun Lu
- Department of Orthopedics, Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Department of Orthopedics, Yancheng First People's Hospital of Jiangsu Province, Yancheng, Jiangsu, China
| | - Guoyou Zou
- Department of Orthopedics, Yancheng First People's Hospital of Jiangsu Province, Yancheng, Jiangsu, China
| | - Duanrong Wu
- Department of Orthopedics, Yancheng First People's Hospital of Jiangsu Province, Yancheng, Jiangsu, China
| | - Yi Cheng
- Department of Orthopedics, Yancheng First People's Hospital of Jiangsu Province, Yancheng, Jiangsu, China
| | - Yongming Sun
- Department of Orthopedics, Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
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21
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Li B, Hu Y, Chen Y, Liu K, Rong K, Hua Q, Fu S, Yang X, Zhou T, Cheng X, Zhang K, Zhao J. Homoplantaginin alleviates intervertebral disc degeneration by blocking the NF-κB/MAPK pathways via binding to TAK1. Biochem Pharmacol 2024; 226:116389. [PMID: 38914318 DOI: 10.1016/j.bcp.2024.116389] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/23/2024] [Revised: 06/20/2024] [Accepted: 06/21/2024] [Indexed: 06/26/2024]
Abstract
Intervertebral disc degeneration (IVDD) is a common degenerative disease which is closely related to low back pain (LBP) and brings huge economic and social burdens. In this study, we explored the therapeutic effects of Homoplantaginin (Hom) for IVDD due to its convincing anti-inflammatory and antioxidant functions. TNF-α was used to simulate the inflammatory environment for nucleus pulposus (NP) cells in vitro. We verified that Hom could alleviate the TNF-α-induced inflammation and disturbance of ECM homeostasis through blocking the NF-κB/MAPK signaling pathways. Subsequently, we screened the binding targets of Hom and confirmed that Hom could directly bind to TAK1 and inhibit its phosphorylation to down-regulate the inflammation-related pathways. The therapeutic effects of Hom on IVDD were further validated through a needle puncture rat model in vivo. Overall, Hom was a promising small molecule for IVDD early intervention, possessing huge clinical translational value.
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Affiliation(s)
- Baixing Li
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Yibin Hu
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Yan Chen
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Kexin Liu
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Kewei Rong
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Qi Hua
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Shaotian Fu
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Xiao Yang
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Tangjun Zhou
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Xiaofei Cheng
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China
| | - Kai Zhang
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China.
| | - Jie Zhao
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011, PR China.
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Yan X, Ding JY, Zhang RJ, Zhang HQ, Kang L, Jia CY, Liu XY, Shen CL. FSTL1 Accelerates Nucleus Pulposus Cell Senescence and Intervertebral Disc Degeneration Through TLR4/NF-κB Pathway. Inflammation 2024; 47:1229-1247. [PMID: 38316670 DOI: 10.1007/s10753-024-01972-0] [Citation(s) in RCA: 9] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/02/2023] [Revised: 01/08/2024] [Accepted: 01/08/2024] [Indexed: 02/07/2024]
Abstract
Intervertebral disc degeneration (IVDD) is a major contributor to low back pain (LBP), and inflammatory factors play crucial roles in its pathogenesis. Follistatin-like 1 (FSTL1) has been reported to induce an inflammatory response in chondrocytes, microglia and preadipocytes, but its role in the pathogenesis of nucleus pulposus cell (NPC) degeneration remains unclear. In this study, we mainly utilized an acidosis-induced NPC degeneration model and a rabbit puncture IVDD model to investigate the role of FSTL1 in IVDD both in vitro and in vivo. We confirmed that FSTL1 expression significantly increased in nucleus pulposus (NP) tissues from IVDD patients and rabbit puncture IVDD models. The expression levels of FSTL1 were significantly increased in all three models of NPC degeneration under harsh microenvironments. In addition, recombinant human FSTL1 (rh-FSTL1) was found to upregulate the expression of p16 and p21, increase the number of senescence-associated β-galactosidase (SA-β-gal)-positive cells, induce senescence-related secretory phenotypes (SASP), and downregulate extracellular matrix (ECM) protein expressions, leading to an imbalance in ECM metabolism destructions. Conversely, silencing of FSTL1 by small interfering RNA (siRNA) ameliorated senescence of NPCs associated with inflammation in IVDD. Furthermore, Toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) pathway plays a crucial role in regulating NPC senescence through FSTL1 regulation. Inhibition of TLR4 expression partly reversed the effects of rh-FSTL1 on NPC senescence-associated inflammation. Finally, rabbit IVDD model experiments demonstrated that the specific FSTL1 siRNA markedly repressed the development of IVDD. These findings may offer a therapeutic approach for mitigating inflammation-induced senescence associated with IVDD.
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Affiliation(s)
- Xu Yan
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Jing-Yu Ding
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Ren-Jie Zhang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Hua-Qing Zhang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Liang Kang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Chong-Yu Jia
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Xiao-Ying Liu
- School of Life Sciences, Anhui Medical University, 81 Meishan Road, Hefei, 230032, Anhui, China.
| | - Cai-Liang Shen
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China.
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China.
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Zhang K, Du L, Li Z, Huo Z, Shen L, Gao S, Jia Y, Zhu M, Xu B. M2 Macrophage-Derived Small Extracellular Vesicles Ameliorate Pyroptosis and Intervertebral Disc Degeneration. Biomater Res 2024; 28:0047. [PMID: 38952714 PMCID: PMC11214826 DOI: 10.34133/bmr.0047] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2024] [Accepted: 05/13/2024] [Indexed: 07/03/2024] Open
Abstract
Intervertebral discs (IVDs) have a limited self-regenerative capacity and current strategies for IVD regeneration are unsatisfactory. Recent studies showed that small extracellular vesicles derived from M2 macrophage cells (M2-sEVs) inhibited inflammation by delivery of various bioactive molecules to recipient cells, which indicated that M2-sEVs may offer a therapeutic strategy for the repair of IVDs. Herein, we investigated the roles and mechanisms of M2-sEVs on IVD regeneration. The in vitro results demonstrated that M2-sEVs inhibited pyroptosis, preserved cellular viability, and promoted migration of nucleus pulposus cells (NPCs). Bioinformatics analysis and verification experiments of microRNA (miR) expression showed that miR-221-3p was highly expressed in M2-sEVs. The mechanism of action was explored and indicated that M2-sEVs inhibited pyroptosis of NPCs through transfer of miR-221-3p, which suppressed the expression levels of phosphatase and tensin homolog and NOD-, LRR-, and pyrin domain-containing protein 3. Moreover, we fabricated decellularized ECM-hydrogel (dECM) for sustained release of M2-sEVs, which exhibited biocompatibility and controlled release properties. The in vivo results revealed that dECM-hydrogel containing M2-sEVs (dECM/M2-sEVs) delayed the degeneration of intervertebral disc degeneration (IDD) models. In addition to demonstrating a promising therapeutic for IDD, this study provided valuable data for furthering the understanding of the roles and mechanisms of M2-sEVs in IVD regeneration.
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Affiliation(s)
- Kaihui Zhang
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital,
Tianjin University, Tianjin 300211, China
| | - Lilong Du
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital,
Tianjin University, Tianjin 300211, China
| | - Zhenhua Li
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital,
Tianjin University, Tianjin 300211, China
| | - Zhenxin Huo
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital,
Tianjin University, Tianjin 300211, China
| | - Li Shen
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital,
Tianjin University, Tianjin 300211, China
| | - Shan Gao
- First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China
| | - Yiming Jia
- Department of Stomatology, Chifeng Municipal Hospital, Chifeng, Inner Mongolia 024000, China
| | - Meifeng Zhu
- College of Life Sciences, Key Laboratory of Bioactive Materials (Ministry of Education),
Nankai University, Tianjin 300071, China
| | - Baoshan Xu
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital,
Tianjin University, Tianjin 300211, China
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24
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Jiang H, Qin H, Yang Q, Huang L, Liang X, Wang C, Moro A, Xu S, Wei Q. Effective delivery of miR-150-5p with nucleus pulposus cell-specific nanoparticles attenuates intervertebral disc degeneration. J Nanobiotechnology 2024; 22:292. [PMID: 38802882 PMCID: PMC11129471 DOI: 10.1186/s12951-024-02561-x] [Citation(s) in RCA: 4] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/15/2024] [Accepted: 05/16/2024] [Indexed: 05/29/2024] Open
Abstract
BACKGROUND The use of gene therapy to deliver microRNAs (miRNAs) has gradually translated to preclinical application for the treatment of intervertebral disc degeneration (IDD). However, the effects of miRNAs are hindered by the short half-life time and the poor cellular uptake, owing to the lack of efficient delivery systems. Here, we investigated nucleus pulposus cell (NPC) specific aptamer-decorated polymeric nanoparticles that can load miR-150-5p for IDD treatment. METHODS The role of miR-150-5p during disc development and degeneration was examined by miR-150-5p knockout (KO) mice. Histological analysis was undertaken in disc specimens. The functional mechanism of miR-150-5p in IDD development was investigated by qRT-PCR assay, Western blot, coimmunoprecipitation and immunofluorescence. NPC specific aptamer-decorated nanoparticles was designed, and its penetration, stability and safety were evaluated. IDD progression was assessed by radiological analysis including X-ray and MRI, after the annulus fibrosus needle puncture surgery with miR-150-5p manipulation by intradiscal injection of nanoparticles. The investigations into the interaction between aptamer and receptor were conducted using mass spectrometry, molecular docking and molecular dynamics simulations. RESULTS We investigated NPC-specific aptamer-decorated polymeric nanoparticles that can bind to miR-150-5p for IDD treatment. Furthermore, we detected that nanoparticle-loaded miR-150-5p inhibitors alleviated NPC senescence in vitro, and the effects of the nanoparticles were sustained for more than 3 months in vivo. The microenvironment of NPCs improves the endo/lysosomal escape of miRNAs, greatly inhibiting the secretion of senescence-associated factors and the subsequent degeneration of NPCs. Importantly, nanoparticles delivering miR-150-5p inhibitors attenuated needle puncture-induced IDD in mouse models by targeting FBXW11 and inhibiting TAK1 ubiquitination, resulting in the downregulation of NF-kB signaling pathway activity. CONCLUSIONS NPC-targeting nanoparticles delivering miR-150-5p show favorable therapeutic efficacy and safety and may constitute a promising treatment for IDD.
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Affiliation(s)
- Hua Jiang
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China.
- Department of Orthopaedic Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China.
| | - Hongyu Qin
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Qinghua Yang
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Longao Huang
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Xiao Liang
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Congyang Wang
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Abu Moro
- Department of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Sheng Xu
- Research Centre for Regenerative Medicine, Guangxi Engineering Center in Biomedical Material for Tissue and Organ Regeneration, Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China
| | - Qingjun Wei
- Department of Orthopaedic Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People's Republic of China.
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25
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Chen X, Chen K, Hu J, Dong Y, Zheng M, Jiang J, Hu Q, Zhang W. Palmitic acid induces lipid droplet accumulation and senescence in nucleus pulposus cells via ER-stress pathway. Commun Biol 2024; 7:539. [PMID: 38714886 PMCID: PMC11076507 DOI: 10.1038/s42003-024-06248-9] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2023] [Accepted: 04/25/2024] [Indexed: 05/12/2024] Open
Abstract
Intervertebral disc degeneration (IDD) is a highly prevalent musculoskeletal disorder affecting millions of adults worldwide, but a poor understanding of its pathogenesis has limited the effectiveness of therapy. In the current study, we integrated untargeted LC/MS metabolomics and magnetic resonance spectroscopy data to investigate metabolic profile alterations during IDD. Combined with validation via a large-cohort analysis, we found excessive lipid droplet accumulation in the nucleus pulposus cells of advanced-stage IDD samples. We also found abnormal palmitic acid (PA) accumulation in IDD nucleus pulposus cells, and PA exposure resulted in lipid droplet accumulation and cell senescence in an endoplasmic reticulum stress-dependent manner. Complementary transcriptome and proteome profiles enabled us to identify solute carrier transporter (SLC) 43A3 involvement in the regulation of the intracellular PA level. SLC43A3 was expressed at low levels and negatively correlated with intracellular lipid content in IDD nucleus pulposus cells. Overexpression of SLC43A3 significantly alleviated PA-induced endoplasmic reticulum stress, lipid droplet accumulation and cell senescence by inhibiting PA uptake. This work provides novel integration analysis-based insight into the metabolic profile alterations in IDD and further reveals new therapeutic targets for IDD treatment.
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Affiliation(s)
- Xi Chen
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China
| | - Kun Chen
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China
| | - Jun Hu
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China
| | - Yijun Dong
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China
| | - Menglong Zheng
- Department of Radiology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China
| | - Jiang Jiang
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China
| | - Qingsong Hu
- Department of Hepatobiliary Surgery, Anhui Province Key Laboratory of Hepatopancreatobiliary Surgery, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China.
| | - Wenzhi Zhang
- Department of Orthopedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230001, China.
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26
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Tao X, Xue F, Xu J, Wang W. Platelet-rich plasma-derived extracellular vesicles inhibit NF-κB/NLRP3 pathway-mediated pyroptosis in intervertebral disc degeneration via the MALAT1/microRNA-217/SIRT1 axis. Cell Signal 2024; 117:111106. [PMID: 38373669 DOI: 10.1016/j.cellsig.2024.111106] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/17/2023] [Revised: 01/26/2024] [Accepted: 02/15/2024] [Indexed: 02/21/2024]
Abstract
BACKGROUND Intervertebral disc degeneration (IDD) is a main contributor to lower back pain, and compression stress-induced apoptosis of nucleus pulposus (NP) cells and extracellular matrix (ECM) degradation has been implicated in the IDD progression. The functions of platelet-rich plasma (PRP)-derived extracellular vesicles (PRP-EVs) in regulating these biological processes remain unclear in IDD. Here, we aimed to investigate the key role of long noncoding RNA (lncRNA) MALAT1 incorporated in PRP-EVs in IDD. METHODS Tert-butyl hydroperoxide (TBHP)-induced damage in NP cells was treated with PRP-EVs extracted from healthy volunteers, followed by MTT, EdU, TUNEL, and Western blot assays. IDD mice were also treated with PRP-EVs. Histomorphological and pathological changes were evaluated. The pyroptosis of cells and the degradation of ECM were detected by ELISA and immunohistochemistry. We screened the differentially expressed lncRNAs in NP cells after PRP-EVs treatment by microarray analysis. The downstream targets of MALAT1 in NP cells were predicted and validated by rescue experiments. FINDINGS TBHP induction reduced cell proliferation and exacerbated pyroptosis and ECM degradation, and PRP-EVs inhibited TBHP-induced cell damage. PRP-EVs-treated mice with IDD had reduced Thompson scores, increased NP tissue content, and restored ECM. PRP-EVs upregulated MALAT1 expression in vivo and in vitro, whereas MALAT1 downregulation exacerbated NP cell pyroptosis and ECM degradation. MALAT1 upregulated SIRT1 expression by downregulating microRNA (miR)-217 in NP cells. SIRT1 blocked the NF-κB/NLRP3 pathway-mediated pyroptosis, thereby alleviating IDD. INTERPRETATION PRP-EVs deliver MALAT1 to regulate miR-217/SIRT1, thereby controlling NP cell pyroptosis in IDD.
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Affiliation(s)
- Xueqiang Tao
- Department of Orthopaedics, the First Affiliated Hospital of Harbin Medical University, Harbin 150000, Heilongjiang, China; Department of Orthopaedics, The Fourth Hospital of BaoTou, Baotou 014030, Inner Mongolia, China
| | - Fen Xue
- Department of Obstetrics and Gynecology, The Fourth Hospital of BaoTou, Baotou 014030, Inner Mongolia, China
| | - Jiayuan Xu
- Department of Orthopaedics, the First Affiliated Hospital of Harbin Medical University, Harbin 150000, Heilongjiang, China
| | - Wenbo Wang
- Department of Orthopaedics, the First Affiliated Hospital of Harbin Medical University, Harbin 150000, Heilongjiang, China.
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27
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Sun K, Yan C, Dai X, Shi Y, Li F, Chen L, Sun J, Chen Y, Shi J. Catalytic Nanodots-Driven Pyroptosis Suppression in Nucleus Pulposus for Antioxidant Intervention of Intervertebral Disc Degeneration. ADVANCED MATERIALS (DEERFIELD BEACH, FLA.) 2024; 36:e2313248. [PMID: 38299823 DOI: 10.1002/adma.202313248] [Citation(s) in RCA: 18] [Impact Index Per Article: 18.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/06/2023] [Revised: 01/17/2024] [Indexed: 02/02/2024]
Abstract
Low back pain resulting from intervertebral disc degeneration (IVDD) is a prevalent global concern; however, its underlying mechanism remains elusive. Single-cell sequencing analyses revealed the critical involvement of pyroptosis in IVDD. Considering the involvement of reactive oxygen species (ROS) as the primary instigator of pyroptosis and the lack of an efficient intervention approach, this study developed carbonized Mn-containing nanodots (MCDs) as ROS-scavenging catalytic biomaterials to suppress pyroptosis of nucleus pulposus (NP) cells to efficiently alleviate IVDD. Catalytic MCDs have superior efficacy in scavenging intracellular ROS and rescuing homeostasis in the NP microenvironment compared with N-acetylcysteine, a classical antioxidant. The data validates that pyroptosis plays a vital role in mediating the protective effects of catalytic MCDs against oxidative stress. Systematic in vivo assessments substantiate the effectiveness of MCDs in rescuing a puncture-induced IVDD rat model, further demonstrating their ability to suppress pyroptosis. This study highlights the potential of antioxidant catalytic nanomedicine as a pyroptosis inhibitor and mechanistically unveils an efficient strategy for the treatment of IVDD.
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Affiliation(s)
- Kaiqiang Sun
- Department of Orthopedic Surgery, Changzheng Hospital, Navy Medical University, Shanghai, 200003, P. R. China
- Department of Orthopedics, Naval Medical Center of PLA, Shanghai, 200052, P. R. China
| | - Chen Yan
- Department of Orthopedic Surgery, Changzheng Hospital, Navy Medical University, Shanghai, 200003, P. R. China
| | - Xinyue Dai
- Materdicine Lab, School of Life Sciences, Shanghai University, Shanghai, 200444, P. R. China
| | - Yangyang Shi
- Department of Orthopedic Surgery, Changzheng Hospital, Navy Medical University, Shanghai, 200003, P. R. China
| | - Fudong Li
- Department of Orthopedic Surgery, Changzheng Hospital, Navy Medical University, Shanghai, 200003, P. R. China
| | - Liang Chen
- Materdicine Lab, School of Life Sciences, Shanghai University, Shanghai, 200444, P. R. China
| | - Jingchuan Sun
- Department of Orthopedic Surgery, Changzheng Hospital, Navy Medical University, Shanghai, 200003, P. R. China
| | - Yu Chen
- Materdicine Lab, School of Life Sciences, Shanghai University, Shanghai, 200444, P. R. China
- Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), Wenzhou Institute of Shanghai University, Wenzhou, 325088, P. R. China
| | - Jiangang Shi
- Department of Orthopedic Surgery, Changzheng Hospital, Navy Medical University, Shanghai, 200003, P. R. China
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28
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Chen Z, Chen C, Yang X, Zhou Y, Cao X, Han C, Zhou T, Zhao J, Qin A. Dysfunction of STING Autophagy Degradation in Senescent Nucleus Pulposus Cells Accelerates Intervertebral Disc Degeneration. Int J Biol Sci 2024; 20:2370-2387. [PMID: 38725841 PMCID: PMC11077376 DOI: 10.7150/ijbs.88534] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/27/2023] [Accepted: 03/16/2024] [Indexed: 05/12/2024] Open
Abstract
The pathogenesis of Intervertebral Disc Degeneration (IDD) is complex and multifactorial, with cellular senescence of nucleus pulposus (NP) cells and inflammation playing major roles in the progression of IDD. The stimulator of interferon genes (STING) axis is a key mediator of inflammation during infection, cellular stress, and tissue damage. Here, we present a progressive increase in STING in senescent NP cells with the degradation disorder. The STING degradation function in normal NP cells can prevent IDD. However, the dysfunction of STING degradation through autophagy causes the accumulation and high expression of STING in senescent NP cells as well as inflammation continuous activation together significantly promotes IDD. In senescent NP cells and intervertebral discs (IVDs), we found that STING autophagy degradation was significantly lower than that of normal NP cells and IVDs when STING was activated by 2'3'-cGAMP. Also, the above phenomenon was found in STINGgt/gt, cGAS-/- mice with models of age-induced, lumbar instability-induced IDD as well as found in the rat caudal IVD puncture models. Taken together, we suggested that the promotion of STING autophagy degradation in senescent NP Cells demonstrated a potential therapeutic modality for the treatment of IDD.
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Affiliation(s)
| | | | | | | | | | | | - Tangjun Zhou
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai, 200011, P. R. China
| | - Jie Zhao
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai, 200011, P. R. China
| | - An Qin
- Shanghai Key Laboratory of Orthopedic Implants, Department of Orthopedics, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, 639 Zhizaoju Road, Shanghai, 200011, P. R. China
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29
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Genedy HH, Humbert P, Laoulaou B, Le Moal B, Fusellier M, Passirani C, Le Visage C, Guicheux J, Lepeltier É, Clouet J. MicroRNA-targeting nanomedicines for the treatment of intervertebral disc degeneration. Adv Drug Deliv Rev 2024; 207:115214. [PMID: 38395361 DOI: 10.1016/j.addr.2024.115214] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2023] [Revised: 02/09/2024] [Accepted: 02/14/2024] [Indexed: 02/25/2024]
Abstract
Low back pain stands as a pervasive global health concern, afflicting almost 80% of adults at some point in their lives with nearly 40% attributable to intervertebral disc degeneration (IVDD). As only symptomatic relief can be offered to patients there is a dire need for innovative treatments.Given the accumulating evidence that multiple microRNAs (miRs) are dysregulated during IVDD, they could have a huge potential against this debilitating condition. The way miRs can profoundly modulate signaling pathways and influence several cellular processes at once is particularly exciting to tackle this multifaceted disorder. However, miR delivery encounters extracellular and intracellular biological barriers. A promising technology to address this challenge is the vectorization of miRs within nanoparticles, providing both protection and enhancing their uptake within the scarce target cells of the degenerated IVD. This comprehensive review presents the diverse spectrum of miRs' connection with IVDD and demonstrates their therapeutic potential when vectorized in nanomedicines.
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Affiliation(s)
- Hussein H Genedy
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France
| | - Paul Humbert
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| | - Bilel Laoulaou
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France
| | - Brian Le Moal
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France
| | - Marion Fusellier
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Department of Diagnostic Imaging, CRIP, ONIRIS, College of Veterinary Medicine, Food Science and Engineering, Nantes F-44307, France
| | | | - Catherine Le Visage
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| | - Jérôme Guicheux
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| | - Élise Lepeltier
- Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France; Institut Universitaire de France (IUF), France.
| | - Johann Clouet
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
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30
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Shen J, Lan Y, Ji Z, Liu H. Sirtuins in intervertebral disc degeneration: current understanding. Mol Med 2024; 30:44. [PMID: 38553713 PMCID: PMC10981339 DOI: 10.1186/s10020-024-00811-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/09/2024] [Accepted: 03/20/2024] [Indexed: 04/01/2024] Open
Abstract
BACKGROUND Intervertebral disc degeneration (IVDD) is one of the etiologic factors of degenerative spinal diseases, which can lead to a variety of pathological spinal conditions such as disc herniation, spinal stenosis, and scoliosis. IVDD is a leading cause of lower back pain, the prevalence of which increases with age. Recently, Sirtuins/SIRTs and their related activators have received attention for their activity in the treatment of IVDD. In this paper, a comprehensive systematic review of the literature on the role of SIRTs and their activators on IVDD in recent years is presented. The molecular pathways involved in the regulation of IVDD by SIRTs are summarized, and the effects of SIRTs on senescence, inflammatory responses, oxidative stress, and mitochondrial dysfunction in myeloid cells are discussed with a view to suggesting possible solutions for the current treatment of IVDD. PURPOSE This paper focuses on the molecular mechanisms by which SIRTs and their activators act on IVDD. METHODS A literature search was conducted in Pubmed and Web of Science databases over a 13-year period from 2011 to 2024 for the terms "SIRT", "Sirtuin", "IVDD", "IDD", "IVD", "NP", "Intervertebral disc degeneration", "Intervertebral disc" and "Nucleus pulposus". RESULTS According to the results, SIRTs and a large number of activators showed positive effects against IVDD.SIRTs modulate autophagy, myeloid apoptosis, oxidative stress and extracellular matrix degradation. In addition, they attenuate inflammatory factor-induced disc damage and maintain homeostasis during disc degeneration. Several clinical studies have reported the protective effects of some SIRTs activators (e.g., resveratrol, melatonin, honokiol, and 1,4-dihydropyridine) against IVDD. CONCLUSION The fact that SIRTs and their activators play a hundred different roles in IVDD helps to better understand their potential to develop further treatments for IVDD. NOVELTY This review summarizes current information on the mechanisms of action of SIRTs in IVDD and the challenges and limitations of translating their basic research into therapy.
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Affiliation(s)
- Jianlin Shen
- Department of Orthopaedics, Affiliated Hospital of Putian University, Putian, 351100, Fujian, China
- Central Laboratory, Affiliated Hospital of Putian University, Putian, 351100, Fujian, China
| | - Yujian Lan
- School of Integrated Traditional Chinese and Western Medicine, Southwest Medical University, Luzhou, 646000, Sichuan, China
| | - Ziyu Ji
- School of Integrated Traditional Chinese and Western Medicine, Southwest Medical University, Luzhou, 646000, Sichuan, China.
| | - Huan Liu
- Department of Orthopaedics, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou, 646000, Sichuan, China.
- The Third People's Hospital of Longmatan District, Luzhou, 646000, Sichuan, China.
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Ding JY, Yan X, Zhang RJ, Zhang HQ, Kang L, Jia CY, Thorne RF, Liu XY, Shen CL. Diagnostic value of serum COMP and ADAMTS7 for intervertebral disc degeneration. Eur J Med Res 2024; 29:196. [PMID: 38528617 PMCID: PMC10962093 DOI: 10.1186/s40001-024-01784-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2023] [Accepted: 03/11/2024] [Indexed: 03/27/2024] Open
Abstract
OBJECTIVE Intervertebral disc degeneration (IVDD) is a major cause of morbidity and disability. Our study aimed to investigate the potential of cartilage oligomeric matrix protein (COMP) and ADAMTS7 (A disintegrin and metalloproteinases with thrombospondin motifs 7) as biomarkers for IVDD together with their functional relationship. METHODS IVD tissues and peripheral blood samples were collected from IVDD rabbit models over 1-4 weeks. Tissues and blood samples were also collected from clinical patients those were stratified into four equal groups according to Pfirrmann IVDD grading (I-V) with baseline data collected for each participant. COMP and ADAMTS7 expression were analyzed and biomarker characteristics were assessed using linear regression and receiver operating curve (ROC) analyses. RESULTS COMP and ADAMTS7 expression increased in tissues and serum during IVDD progression. Serum COMP (sCOMP) and serum ADAMTS7 (sADAMTS7) levels increased in a time-dependent manner following IVD damage in the rabbit model while significant positive correlations were detected between sCOMP and sADAMTS7 and Pfirrmann grade in human subjects. ROC analysis showed that combining sCOMP and sADAMTS7 assay results produced an improved diagnostic measure for IVDD compared to individual sCOMP or sADAMTS7 tests. In vitro assays conducted on human cell isolates revealed that COMP prevented extracellular matrix degradation and antagonized ADAMTS7 expression although this protective role was uncoupled under microenvironmental conditions mimicking IVDD. CONCLUSIONS Increases in circulating COMP and ADAMTS7 correlate with IVDD progression and may play regulatory roles. Assays for sCOMP and/or sADAMTS7 levels can discriminate between healthy subjects and IVDD patients, warranting further clinical assessment.
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Affiliation(s)
- Jing-Yu Ding
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Xu Yan
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Ren-Jie Zhang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Hua-Qing Zhang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Liang Kang
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Chong-Yu Jia
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China
| | - Rick F Thorne
- Henan International Joint Laboratory of Non-Coding RNA and Metabolism in Cancer, Henan Provincial Key Laboratory of Long Non-Coding RNA and Cancer Metabolism, Translational Research Institute of Henan Provincial People's Hospital and People's Hospital of Zhengzhou University, Zhengzhou, Henan, China
| | - Xiao-Ying Liu
- School of Life Sciences, Anhui Medical University, 81 Meishan Road, Hefei, 230032, Anhui, China.
- Henan International Joint Laboratory of Non-Coding RNA and Metabolism in Cancer, Henan Provincial Key Laboratory of Long Non-Coding RNA and Cancer Metabolism, Translational Research Institute of Henan Provincial People's Hospital and People's Hospital of Zhengzhou University, Zhengzhou, Henan, China.
| | - Cai-Liang Shen
- Department of Orthopedics and Spine Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China.
- Laboratory of Spinal and Spinal Cord Injury Regeneration and Repair, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China.
- Anhui Province Research Center for the Clinical Application of Digital Medical Technology, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, Anhui, China.
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Li Y, Wang B, Sun W, Kong C, Ding J, Hu F, Li J, Chen X, Lu S. Construction of circ_0071922-miR-15a-5p-mRNA network in intervertebral disc degeneration by RNA-sequencing. JOR Spine 2024; 7:e1275. [PMID: 38222808 PMCID: PMC10782064 DOI: 10.1002/jsp2.1275] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/21/2023] [Revised: 06/25/2023] [Accepted: 07/19/2023] [Indexed: 01/16/2024] Open
Abstract
Background Low back pain (LBP) is the main factor of global disease burden. Intervertebral disc degeneration (IVDD) has long been known as the leading reason of LBP. Increasing studies have verified that circular RNAs (circRNAs)-microRNAs (miRNAs)-mRNAs network is widely involved in the pathological processes of IVDD. However, no study was made to demonstrate the circRNAs-mediated ferroptosis, oxidative stress, extracellular matrix metabolism, and immune response in IVDD. Methods We collected 3 normal and 3 degenerative nucleus pulposus tissues to conduct RNA-sequencing to identify the key circRNAs and miRNAs in IVDD. Bioinformatics analysis was then conducted to construct circRNAs-miRNAs-mRNAs interaction network associated with ferroptosis, oxidative stress, extracellular matrix metabolism, and immune response. We also performed animal experiments to validate the therapeutic effects of key circRNAs in IVDD. Results We found that circ_0015435 was most obviously upregulated and circ_0071922 was most obviously downregulated in IVDD using RNA-sequencing. Then we observed that hsa-miR-15a-5p was the key downstream of circ_0071922, and hsa-miR-15a-5p was the top upregulated miRNA in IVDD. Bioinformatics analysis was conducted to predict that 56 immunity-related genes, 29 ferroptosis-related genes, 23 oxidative stress-related genes and 8 ECM-related genes are the targets mRNAs of hsa-miR-15a-5p. Then we constructed a ceRNA network encompassing 24 circRNAs, 6 miRNAs, and 101 mRNAs. Additionally, we demonstrated that overexpression of circ_0071922 can alleviate IVDD progression in a rat model. Conclusions The findings of this study suggested that circ_0071922-miR-15a-5p-mRNA signaling network might affect IVDD by modulating the nucleus pulposus cells ferroptosis, oxidative stress, ECM metabolism, and immune response, which is an effective therapeutic targets of IVDD.
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Affiliation(s)
- Yongjin Li
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
| | - Baobao Wang
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
| | - Wenzhi Sun
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
| | - Chao Kong
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
| | - Junzhe Ding
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
| | - Feng Hu
- Spine Center, Department of Orthopaedics, The First Affiliated Hospital of USTC, Division of Life Sciences and MedicineUniversity of Science and Technology of ChinaHefeiAnhuiChina
| | - Jianhua Li
- Department of OrthopedicsTianjin Haihe HospitalTianjinChina
| | - Xiaolong Chen
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
| | - Shibao Lu
- Department of OrthopedicsXuanwu Hospital, Capital Medical UniversityBeijingChina
- National Clinical Research Center for Geriatric DiseasesBeijingChina
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Song H, Guo C, Wu Y, Liu Y, Kong Q, Wang Y. Therapeutic factors and biomaterial-based delivery tools for degenerative intervertebral disc repair. Front Cell Dev Biol 2024; 12:1286222. [PMID: 38374895 PMCID: PMC10875104 DOI: 10.3389/fcell.2024.1286222] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2023] [Accepted: 01/15/2024] [Indexed: 02/21/2024] Open
Abstract
Intervertebral disc degeneration (IDD) is the main cause of low back pain (LBP), which significantly impacts global wellbeing and contributes to global productivity declines. Conventional treatment approaches, encompassing conservative and surgical interventions, merely serve to postpone the advancement of IDD without offering a fundamental reversal. Consequently, there is an urgent demand for an effective approach to prevent the progression of IDD. Recent investigations focusing on the treatment of IDD utilizing diverse bioactive substances integrated within various biomaterials have exhibited promising outcomes. Various bioactive substances, encompassing conventional small molecule drugs, small molecule nucleic acids, and cell therapies, exhibit distinct capacities for repairing IDD. Additionally, various biological material delivery systems, such as nano micelles, microspheres, and hydrogels, possess diverse biological and release characteristics. Consequently, these diverse materials and drugs hold promise for advancing the treatment of IDD. This article aims to provide a concise overview of the IDD process and investigate the research advancements in biomaterials and bioactive substances for IDD treatment, delving into their mechanisms.
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Affiliation(s)
| | | | | | | | - Qingquan Kong
- Department of Orthopedic Surgery and Orthopedic Research Institute, West China Hospital, Sichuan University, Chengdu, Sichuan, China
| | - Yu Wang
- Department of Orthopedic Surgery and Orthopedic Research Institute, West China Hospital, Sichuan University, Chengdu, Sichuan, China
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Yang S, Jing S, Wang S, Jia F. From drugs to biomaterials: a review of emerging therapeutic strategies for intervertebral disc inflammation. Front Cell Infect Microbiol 2024; 14:1303645. [PMID: 38352058 PMCID: PMC10861683 DOI: 10.3389/fcimb.2024.1303645] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2023] [Accepted: 01/12/2024] [Indexed: 02/16/2024] Open
Abstract
Chronic low back pain (LBP) is an increasingly prevalent issue, especially among aging populations. A major underlying cause of LBP is intervertebral disc degeneration (IDD), often triggered by intervertebral disc (IVD) inflammation. Inflammation of the IVD is divided into Septic and Aseptic inflammation. Conservative therapy and surgical treatment often fail to address the root cause of IDD. Recent advances in the treatment of IVD infection and inflammation range from antibiotics and small-molecule drugs to cellular therapies, biological agents, and innovative biomaterials. This review sheds light on the complex mechanisms of IVD inflammation and physiological and biochemical processes of IDD. Furthermore, it provides an overview of recent research developments in this area, intending to identify novel therapeutic targets and guide future clinical strategies for effectively treating IVD-related conditions.
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Affiliation(s)
- Shuhan Yang
- Department of Orthopedics, Third Hospital of Shanxi Medical University, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Taiyuan, China
| | - Shaoze Jing
- Department of Orthopedics, Third Hospital of Shanxi Medical University, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Taiyuan, China
| | - Shanxi Wang
- Department of Spine Surgery, Honghui Hospital, Xi’an Jiaotong University, Xi’an, China
| | - Fajing Jia
- Department of General Practice, Third Hospital of Shanxi Medical University, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Taiyuan, China
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Chen F, Lei L, Chen S, Zhao Z, Huang Y, Jiang G, Guo X, Li Z, Zheng Z, Wang J. Serglycin secreted by late-stage nucleus pulposus cells is a biomarker of intervertebral disc degeneration. Nat Commun 2024; 15:47. [PMID: 38167807 PMCID: PMC10761730 DOI: 10.1038/s41467-023-44313-9] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2022] [Accepted: 12/05/2023] [Indexed: 01/05/2024] Open
Abstract
Intervertebral disc degeneration is a natural process during aging and a leading cause of lower back pain. Here, we generate a comprehensive atlas of nucleus pulposus cells using single-cell RNA-seq analysis of human nucleus pulposus tissues (three males and four females, age 41.14 ± 18.01 years). We identify fibrotic late-stage nucleus pulposus cells characterized by upregulation of serglycin expression which facilitate the local inflammatory response by promoting the infiltration of inflammatory cytokines and macrophages. Finally, we discover that daphnetin, a potential serglycin ligand, substantially mitigates the local inflammatory response by downregulating serglycin expression in an in vivo mouse model, thus alleviating intervertebral disc degeneration. Taken together, we identify late-stage nucleus pulposus cells and confirm the potential mechanism by which serglycin regulates intervertebral disc degeneration. Our findings indicate that serglycin is a latent biomarker of intervertebral disc degeneration and may contribute to development of diagnostic and therapeutic strategies.
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Affiliation(s)
- Fan Chen
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
- Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China
| | - Linchuan Lei
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
- Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China
| | - Shunlun Chen
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
| | - Zhuoyang Zhao
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
- Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China
| | - Yuming Huang
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
| | - Guowei Jiang
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
- Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China
| | - Xingyu Guo
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
| | - Zemin Li
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China
| | - Zhaomin Zheng
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China.
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China.
| | - Jianru Wang
- Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, P.R. China.
- Guangdong Province Key Laboratory of Orthopaedics and Traumatology, Guangzhou, 510080, P.R. China.
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Jiang C, Liu Y, Zhao W, Yang Y, Ren Z, Wang X, Hao D, Du H, Yin S. microRNA-365 attenuated intervertebral disc degeneration through modulating nucleus pulposus cell apoptosis and extracellular matrix degradation by targeting EFNA3. J Cell Mol Med 2024; 28:e18054. [PMID: 38009813 PMCID: PMC10826450 DOI: 10.1111/jcmm.18054] [Citation(s) in RCA: 2] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/25/2023] [Revised: 10/30/2023] [Accepted: 11/14/2023] [Indexed: 11/29/2023] Open
Abstract
This present study is aimed to investigate the role of microRNA-365 (miR-365) in the development of intervertebral disc degeneration (IDD). Nucleus pulposus (NP) cells were transfected by miR-365 mimic and miR-365 inhibitor, respectively. Concomitantly, the transfection efficiency and the expression level of miRNA were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Meanwhile, NP cells apoptosis was measured through propidium iodide (PI)-AnnexinV-fluorescein isothiocyanate (FITC) apoptosis detection kit. Subsequently, immunofluorescence (IF) staining was performed to assess the expression of collagen II, aggrecan and matrix metalloproteinase 13 (MMP-13). In addition, bioinformatic prediction and Luciferase reporter assay were used to reveal the target gene of miR-365. Finally, we isolated the primary NP cells from rats and injected NP-miR-365 in rat IDD models. The results showed that overexpression of miR-365 could effectively inhibit NP cells apoptosis and MMP-13 expression and upregulate the expression of collagen II and aggrecan. Conversely, suppression of miR-365 enhanced NP cell apoptosis and elevated MMP-13 expression, but decreased the expression of collagen II and aggrecan. Moreover, the further data demonstrated that miR-365 mediated NP cell degradation through targeting ephrin-A3 (EFNA3). In addition, the cells apoptosis and catabolic markers were increased in NP cells when EFNA3 upregulated. More importantly, the vivo data supported that miR-365-NP cells injection ameliorated IDD in rats models. miR-365 could alleviate the development of IDD by regulating NP cell apoptosis and ECM degradation, which is likely mediated by targeting EFNA3. Therefore, miR-365 may be a promising therapeutic avenue for treatment IDD through EFNA3.
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Affiliation(s)
- Chao Jiang
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Youjun Liu
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Weigong Zhao
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Yimin Yang
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Zhiwei Ren
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Xiaohui Wang
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
- Department of Developmental GeneticsMax Planck Institute for Heart and Lung ResearchBad NauheimGermany
| | - Dingjun Hao
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Heng Du
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
| | - Si Yin
- Department of Orthopedic SurgeryThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anChina
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Li XC, Wang W, Jiang C, Chen YL, Chen JH, Zhang ZW, Luo SJ, Chen RC, Mo PF, Zhong ML, Shi JY, Huang CM, Chen Q, Wu YH. CD206 + M2-like macrophages protect against intervertebral disc degeneration partially by targeting R-spondin-2. Osteoarthritis Cartilage 2024; 32:66-81. [PMID: 37802465 DOI: 10.1016/j.joca.2023.09.010] [Citation(s) in RCA: 1] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/20/2023] [Revised: 08/31/2023] [Accepted: 09/25/2023] [Indexed: 10/10/2023]
Abstract
OBJECTIVE This study aimed to explore the specific function of M2 macrophages in intervertebral disc degeneration (IDD). METHODS Intervertebral disc (IVD) samples from normal (n = 4) and IDD (n = 6) patients were collected, and the expression of M2-polarized macrophage marker, CD206, was investigated using immunohistochemical staining. Nucleus pulposus cells (NPCs) in a TNF-α environment were obtained, and a mouse caudal IVD puncture model was established. Mice with Rheb deletions, specifically in the myeloid lineage, were generated and subjected to surgery-induced IDD. IDD-induced damage and cell apoptosis were measured using histological scoring, X-ray imaging, immunohistochemical staining, and TdT-mediated dUTP nick end labeling (TUNEL) assay. Finally, mice and NPCs were treated with R-spondin-2 (Rspo2) or anti-Rspo2 to investigate the role of Rspo2 in IDD. RESULTS Accumulation of CD206 in human and mouse IDD tissues was detected. Rheb deletion in the myeloid lineage (RheBcKO) increased the number of CD206+ M2-like macrophages (mean difference 18.6% [15.7-21.6%], P < 0.001), decreased cell apoptosis (mean difference -15.6% [-8.9 to 22.2%], P = 0.001) and attenuated the IDD process in the mouse IDD model. NPCs treated with Rspo2 displayed increased extracellular matrix catabolism and apoptosis; co-culture with a conditioned medium derived from RheBcKO mice inhibited these changes. Anti-Rspo2 treatment in the mouse caudal IVD puncture model exerted protective effects against IDD. CONCLUSIONS Promoting CD206+ M2-like macrophages could reduce Rspo2 secretion, thereby alleviating experimental IDD. Rheb deletion may help M2-polarized macrophages accumulate and attenuate experimental IDD partially by inhibiting Rspo2 production. Hence, M2-polarized macrophages and Rspo2 may serve as therapeutic targets for IDD.
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Affiliation(s)
- Xiao-Chuan Li
- Department of Orthopedic Surgery, Gaozhou People's Hospital, No.89 XiGuan Rd, Gaozhou 525200, Guangdong, China; Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China; Postdoctoral Innovation Practice Base of Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China
| | - Wei Wang
- Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China; Graduate School of Guangdong Medical University, No. 2, Wenming East Road, Zhanjiang 524023, Guangdong, China
| | - Cheng Jiang
- Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China; Graduate School of Guangdong Medical University, No. 2, Wenming East Road, Zhanjiang 524023, Guangdong, China
| | - Yong-Long Chen
- Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China; Graduate School of Guangdong Medical University, No. 2, Wenming East Road, Zhanjiang 524023, Guangdong, China
| | - Jiong-Hui Chen
- Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China; Graduate School of Guangdong Medical University, No. 2, Wenming East Road, Zhanjiang 524023, Guangdong, China
| | - Zhen-Wu Zhang
- Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China; Graduate School of Guangdong Medical University, No. 2, Wenming East Road, Zhanjiang 524023, Guangdong, China
| | - Shao-Jian Luo
- Department of Orthopedic Surgery, Gaozhou People's Hospital, No.89 XiGuan Rd, Gaozhou 525200, Guangdong, China
| | - Rong-Chun Chen
- Department of Spinal Surgery, Ganzhou People's Hospital, No.16 Meiguan Avenue, Ganzhou 341000, Jiangxi, China
| | - Ping-Fan Mo
- Department of Spinal Surgery, Ganzhou People's Hospital, No.16 Meiguan Avenue, Ganzhou 341000, Jiangxi, China
| | - Ming-Liang Zhong
- Department of Spinal Surgery, Ganzhou People's Hospital, No.16 Meiguan Avenue, Ganzhou 341000, Jiangxi, China
| | - Jiang-You Shi
- Department of Spinal Surgery, Ganzhou People's Hospital, No.16 Meiguan Avenue, Ganzhou 341000, Jiangxi, China
| | - Chun-Ming Huang
- Department of Orthopedic Surgery, Gaozhou People's Hospital, No.89 XiGuan Rd, Gaozhou 525200, Guangdong, China; Central Laboratory of Orthopedics, Gaozhou People's Hospital, XiGuan Rd, Gaozhou 525200, China.
| | - Qin Chen
- Department of Spinal Surgery, Ganzhou People's Hospital, No.16 Meiguan Avenue, Ganzhou 341000, Jiangxi, China.
| | - Yao-Hong Wu
- Department of Spinal Surgery, Ganzhou People's Hospital, No.16 Meiguan Avenue, Ganzhou 341000, Jiangxi, China.
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Chen Z, Liao Z, Liu M, Lin F, Chen S, Wang G, Zheng Z, Liu B, Li C, Wang Z, Chen T, Huang H, Liao Q, Cui W. Nucleus Pulposus-Targeting Nanocarriers Facilitate Mirna-Based Therapeutics for Intervertebral Disc Degeneration. Adv Healthc Mater 2023; 12:e2301337. [PMID: 37625164 DOI: 10.1002/adhm.202301337] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/26/2023] [Revised: 07/23/2023] [Indexed: 08/27/2023]
Abstract
Intervertebral disc degeneration (IDD) is a common cause of low back pain. Understanding its molecular mechanisms is the basis for developing specific treatment. To demonstrate that miR-22-3p is critical in the regulation of IDD, miRNA microarray analyses are conducted in conjunction with in vivo and in vitro experiments. The miR-22-3p knockout (KO) mice show a marked decrease in the histological scores. Bioinformatic analysis reveals that miR-22-3p plays a mechanistic role in the development of IDD by targeting SIRT1, which in turn activates the JAK1/STAT3 signaling pathway. This is confirmed by a luciferase reporter assay and western blot analysis. Therapeutically, the delivery of miR-22-3p inhibitors and mimics through the synthesized nanoparticles in the IDD model alleviates and aggravates IDD, respectively. The nanocarriers enhance transportation of miR-22-3p to nucleus pulposus cells, thus enabling the in vivo inhibition of miR-22-3p for therapeutic purposes and consequently promoting the development of miRNA-specific drugs for IDD.
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Affiliation(s)
- Zhonghui Chen
- Orthopaedic Surgery, Fujian Medical University Union Hospital, Fuzhou, Fujian, 350000, China
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
- Orthopaedic Surgery, Renmin Hospital of Wuhan University, Wuhan, Hubei, 430000, China
| | - Zhong Liao
- Orthopaedic Surgery, Fujian Medical University Union Hospital, Fuzhou, Fujian, 350000, China
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Ming Liu
- Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, Fuzhou, Fujian, 350000, China
| | - Fengfei Lin
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Shunyou Chen
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Geng Wang
- Department of Pharmacology, School of Pharmacy, Fujian Provincial Key Laboratory of Natural Medicine Pharmacology, Fujian Medical University, Fuzhou, Fujian, 350000, China
| | - Zhong Zheng
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Boling Liu
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Chaoxiong Li
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Zheqiang Wang
- Department of Sport's Medicine, The Second Affiliated Hospital of Fujian Traditional Chinese Medical University, Fuzhou, Fujian, 350000, China
| | - Tianlai Chen
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Hongzhe Huang
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
| | - Qi Liao
- Orthopaedic Surgery, Renmin Hospital of Wuhan University, Wuhan, Hubei, 430000, China
| | - Weiliang Cui
- Orthopaedic Surgery, Fujian Medical University Union Hospital, Fuzhou, Fujian, 350000, China
- Orthopaedic Surgery, Fuzhou Second Hospital, Fuzhou, Fujian, 350000, China
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Han H, Zhao X, Ma H, Zhang Y, Lei B. Multifunctional injectable hydrogels with controlled delivery of bioactive factors for efficient repair of intervertebral disc degeneration. Heliyon 2023; 9:e21867. [PMID: 38027562 PMCID: PMC10665751 DOI: 10.1016/j.heliyon.2023.e21867] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/16/2023] [Revised: 10/07/2023] [Accepted: 10/30/2023] [Indexed: 12/01/2023] Open
Abstract
Millions of people worldwide suffer from intervertebral disc degeneration (IVDD), which imposes a significant socioeconomic burden on society. There is an urgent clinical demand for more effective treatments for IVDD because conventional treatments can only alleviate the symptoms rather than preventing the progression of IVDD. Hydrogels, a class of elastic biomaterials with good biocompatibility, are promising candidates for intervertebral disc repair and regeneration. In recent years, various hydrogels have been investigated in vitro and in vivo for the repair of intervertebral discs, some of which are ready for clinical testing. This review summarizes the latest findings and developments in using bioactive factors-released bioactive injectable hydrogels for the repair and regeneration of intervertebral discs. It focuses on the analysis and summary of the use of multifunctional injectable hydrogels to delivery bioactive factors (cells, exosomes, growth factors, genes, drugs) for disc regeneration, providing guidance for future study. Finally, we discussed and analyzed the optimal timing for the application of controlled-release hydrogels in the treatment of IVDD to meet the high standards required for intervertebral disc regeneration and precision medicine.
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Affiliation(s)
- Hao Han
- Department of Orthopaedics of the First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, 710061, China
| | - Xiaoming Zhao
- Department of Orthopaedics of the First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, 710061, China
| | - Hongyun Ma
- Department of Orthopaedics of the First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, 710061, China
| | - Yingang Zhang
- Department of Orthopaedics of the First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, 710061, China
| | - Bo Lei
- Department of Orthopaedics of the First Affiliated Hospital, Xi'an Jiaotong University, Xi'an, 710061, China
- Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an, 710000, China
- Fronter Institute of Science and Technology, Xi'an Jiaotong University, Xi'an, 710000, China
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Li Y, Kong C, Wang W, Hu F, Chen X, Xu B, Lu S. Screening of miR-15a-5p as a potential biomarker for intervertebral disc degeneration through RNA-sequencing. Int Immunopharmacol 2023; 123:110717. [PMID: 37597405 DOI: 10.1016/j.intimp.2023.110717] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/15/2023] [Revised: 07/25/2023] [Accepted: 07/25/2023] [Indexed: 08/21/2023]
Abstract
Low back pain (LBP) is a prevalent clinical condition that imposes substantial economic burdens on society. Intervertebral disc degeneration (IVDD) is recognized as a major contributing factor to LBP. Recent studies have highlighted the pivotal role of microRNAs (miRNAs) in regulating the onset and progression of IVDD. Understanding the involvement of miRNAs in IVDD will expand our knowledge of the underlying mechanisms and potentially identify novel therapeutic targets for managing LBP. However, the pathological process of IVDD and the miRNA-mediated pathomechanism in IVDD remain unclear. Herein, we comprehensively analyzed and divided the pathological process of IVDD into three stages based on the analysis by Risbud and colleagues. Results showed that IVDD was especially associated with cell death, oxidative stress, inflammatory and immune response, and extracellular matrix (ECM) metabolism. Subsequently, we obtained human normal and degenerative nucleus pulposus tissues, which were visually confirmed through histological staining techniques such as HE and TUNEL staining. RNA sequencing was then performed on these tissue samples. Additionally, miRNA (GSE116726) and mRNA (GSE56081/GSE70362/GSE23130/GSE34095) datasets were collected from the GEO database. Our analysis revealed that miR-15a-5p was significantly upregulated IVDD, as validated by both RNA sequencing and qRT-PCR experiments. To further refine our findings, bioinformatics analysis was conducted, merging the targets of miR-15a-5p and multiple mRNA datasets, ultimately identifying the overlapping IVDD-associated mRNAs. Notably, many cuproptosis-related genes (CRGs), ferroptosis-related genes, oxidative stress-related genes, and immunity-related genes were potential targets of miR-15a-5p. The miR-15a-5p-mRNA network was constructed using Cytoscape software. Additionally, PPI, functional, and pathway enrichment analyses of the CRGs were also performed. We found that MTF1, one of the CRGs, was highly expressed in IVDD and primarily localized in the nucleus of nucleus pulposus cells. These findings suggest that miR-15a-5p is a potential biomarker in IVDD, and targeting the miR-15a-5p-mRNA signaling pathway may be a promising strategy for treating IVDD diseases.
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Affiliation(s)
- Yongjin Li
- Department of Orthopedics, Xuanwu Hospital, Capital Medical University, No. 45 Changchun Street, Xicheng District, Beijing, China; National Clinical Research Center for Geriatric Diseases, Beijing, China
| | - Chao Kong
- Department of Orthopedics, Xuanwu Hospital, Capital Medical University, No. 45 Changchun Street, Xicheng District, Beijing, China; National Clinical Research Center for Geriatric Diseases, Beijing, China
| | - Wei Wang
- Department of Orthopedics, Xuanwu Hospital, Capital Medical University, No. 45 Changchun Street, Xicheng District, Beijing, China; National Clinical Research Center for Geriatric Diseases, Beijing, China
| | - Feng Hu
- Spine Center, Department of Orthopaedics, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui 230001, China
| | - Xiaolong Chen
- Department of Orthopedics, Xuanwu Hospital, Capital Medical University, No. 45 Changchun Street, Xicheng District, Beijing, China; National Clinical Research Center for Geriatric Diseases, Beijing, China.
| | - Baoshan Xu
- Department of Minimally Invasive Spine Surgery, Tianjin Hospital, 406. No, Jiefangnan Road, Hexi district, Tianjin 300211, China.
| | - Shibao Lu
- Department of Orthopedics, Xuanwu Hospital, Capital Medical University, No. 45 Changchun Street, Xicheng District, Beijing, China; National Clinical Research Center for Geriatric Diseases, Beijing, China.
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Chen Y, Cao X, Pan B, Du H, Li B, Yang X, Chen X, Wang X, Zhou T, Qin A, Zhao C, Zhao J. Verapamil attenuates intervertebral disc degeneration by suppressing ROS overproduction and pyroptosis via targeting the Nrf2/TXNIP/NLRP3 axis in four-week puncture-induced rat models both in vivo and in vitro. Int Immunopharmacol 2023; 123:110789. [PMID: 37579541 DOI: 10.1016/j.intimp.2023.110789] [Citation(s) in RCA: 12] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2023] [Revised: 07/22/2023] [Accepted: 08/09/2023] [Indexed: 08/16/2023]
Abstract
Low back pain is usually caused by intervertebral disc degeneration (IVDD), during which the involvement of oxidation system imbalance and inflammasome activation cannot be neglected. In this study, we aimed to validate the expression level of TXNIP in IVDD and investigate the function and potential mechanism of action of verapamil. TXNIP is upregulated in the degenerate nucleus pulposus in both humans and rats, as well as in tert-butyl hydroperoxide (TBHP)-stimulated nucleus pulposus cells. Administration of verapamil, a classic clinical drug, mitigated the TBHP-induced overproduction of reactive oxygen species and activation of the NLRP3 inflammasome, thus protecting cells from pyroptosis, apoptosis, and extracellular matrix degradation. The Nrf2/TXNIP/NLRP3 axis plays a major role in verapamail-mediated protection. In vivo, a puncture-induced IVDD rat model was constructed, and we found that verapamil delayed the development of IVDD at both the imaging and histological levels. In summary, our results indicate the potential therapeutic effects and mechanisms of action of verapamil in the treatment of IVDD.
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Affiliation(s)
- Yan Chen
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Xiankun Cao
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Bin Pan
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Han Du
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Baixing Li
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Xiao Yang
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Xuzhuo Chen
- Department of Oral Surgery, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Xin Wang
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - Tangjun Zhou
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
| | - An Qin
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China.
| | - Changqing Zhao
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China.
| | - Jie Zhao
- Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China; Shanghai Key Laboratory of Orthopaedic Implants, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, PR China
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Yuan J, Li S, Han Y, Li F, Shi H, Shi W, Cui W. Restoration of miR-328a-5p function curtails hypoxic pulmonary hypertension through a mechanism involving PIN1/GSK3β/β-catenin axis. Int Immunopharmacol 2023; 123:110599. [PMID: 37567011 DOI: 10.1016/j.intimp.2023.110599] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2022] [Revised: 06/19/2023] [Accepted: 07/01/2023] [Indexed: 08/13/2023]
Abstract
Recent evidence has highlighted the involvement of microRNAs (miRs) in hypoxic pulmonary hypertension (PH), which can be induced under hypoxic conditions. We intend to explore whether the miR-328a-5p/PIN1 axis affects hypoxic PH by regulating the GSK3β/β-catenin signaling pathway. The GEO database was retrieved to single out key miRs affecting hypoxic PH. It was observed that downregulation of miR-328a-5p occurred in hypoxia-induced PH samples. The binding affinity between miR-328a-5p to PIN1 was predicted by a bioinformatics tool and verified using a dual luciferase reporter gene assay. Rat primary pulmonary artery smooth muscle cells (PASMCs) were exposed to hypoxia for in vitro cell experiments. miR-328a-5p could target and downregulate PIN1 expression, leading to suppressed GSK3β/β-catenin activation. In addition, GSK3β/β-catenin inactivation curtailed hypoxia-induced vascular inflammatory responses and proliferation and migration in PASMCs in vitro. A hypoxic PH model was established in SD rats to observe the effects of miR-328a-5p on hemodynamic parameters and right heart remodeling. It was demonstrated in vivo that miR-328a-5p downregulated PIN1 expression to suppress GSK3β/β-catenin signaling, thereby reducing the vascular inflammatory response and alleviating disease progression in hypoxia-induced PH rats. The evidence provided by our study highlighted the involvement of miR-328a-5p in the translational suppression of PIN1 and the blockade of the GSK3β/β-catenin signaling pathway, resulting in attenuation of hypoxic PH progression.
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Affiliation(s)
- Jieqing Yuan
- Department of Respiratory and Critical Care Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China
| | - Shanshan Li
- Department of Respiratory and Critical Care Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China
| | - Yu Han
- Department of Respiratory and Critical Care Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China
| | - Fujun Li
- Department of Emergency Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China
| | - Hai Shi
- Department of Respiratory and Critical Care Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China
| | - Weitao Shi
- Department of Critical Care Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China
| | - Wenjie Cui
- Department of Respiratory and Critical Care Medicine, The First People's Hospital of Xuzhou, the Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou 221100, PR China.
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Qingxin S, Kai J, Dandan Z, Linyu J, Xiuyuan C, Yubo F, Kun W, Yingchao H, Hao C, Jie S, Zhi C, Hongxing S. Programmable DNA hydrogel provides suitable microenvironment for enhancing autophagy-based therapies in intervertebral disc degeneration treatment. J Nanobiotechnology 2023; 21:350. [PMID: 37759249 PMCID: PMC10537074 DOI: 10.1186/s12951-023-02109-5] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2023] [Accepted: 09/15/2023] [Indexed: 09/29/2023] Open
Abstract
The pathogenesis of intervertebral disc degeneration (IVDD) is attributed to metabolic dysregulation within the extracellular matrix and heightened apoptosis of nucleus pulposus cells (NPC). Therefore, a potential therapeutic strategy for managing IVDD involves the reestablishment of metabolic equilibrium within the extracellular matrix and the suppression of excessive myeloid cell apoptosis. The microRNA, miR-5590, displays marked differential expression in degenerative nucleus pulposus (NP) tissues and exerts a direct influence on the regulation of DDX5 expression. This, in turn, modulates mammalian target of rapamycin (mTOR) phosphorylation, thereby impacting autophagy and apoptosis. However, ensuring the smooth delivery of miRNA to a specific injury site poses a significant challenge. To address this issue, a multifunctional DNA hydrogel was developed and subsequently loaded with miR-5590 via spherical nucleic acids (SNAs) for the treatment of IVDD. The hydrogel, which exhibits versatility, has the potential to be administered through injection at the site of injury, resulting in a consistent and prolonged release of miR-5590. This leads to the creation of a genetic microenvironment within the NP, which triggers the onset of autophagy in NPCs and subsequently suppresses apoptosis. As a result, this process regulates the metabolic equilibrium within the extracellular matrix, thereby impeding the in vitro and in vivo progression of IVDD. The amalgamation of miRNAs and biomaterials offers a promising therapeutic strategy for the management of IVDD in clinical settings.
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Affiliation(s)
- Song Qingxin
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Jiang Kai
- Department of Ophthalmology and Vision Science, Ear, Nose and Throat Hospital, Shanghai Eye, Fudan University, Shanghai, China
| | - Zheng Dandan
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Jin Linyu
- Department of Orthopedics, Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases, Shanghai Institute of Traumatology and Orthopedics, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, P. R. China
| | - Chen Xiuyuan
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Feng Yubo
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Wang Kun
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Han Yingchao
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Chen Hao
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China
| | - Song Jie
- Department of Instrument Science and Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
- The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou, 310022, Zhejiang, China.
| | - Chen Zhi
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China.
| | - Shen Hongxing
- Department of Spine Surgery Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai, 200127, P. R. China.
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Liang H, Luo R, Li G, Zhang W, Zhu D, Wu D, Zhou X, Tong B, Wang B, Feng X, Wang K, Song Y, Yang C. Lysine methylation of PPP1CA by the methyltransferase SUV39H2 disrupts TFEB-dependent autophagy and promotes intervertebral disc degeneration. Cell Death Differ 2023; 30:2135-2150. [PMID: 37605006 PMCID: PMC10482945 DOI: 10.1038/s41418-023-01210-4] [Citation(s) in RCA: 9] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2022] [Revised: 08/02/2023] [Accepted: 08/08/2023] [Indexed: 08/23/2023] Open
Abstract
Impaired transcription factor EB (TFEB) function and deficient autophagy activity have been shown to aggravate intervertebral disc (IVD) degeneration (IDD), yet the underlying mechanisms remain less clear. Protein posttranslational modifications (PTMs) are critical for determining TFEB trafficking and transcriptional activity. Here, we demonstrate that TFEB activity is controlled by protein methylation in degenerated nucleus pulposus cells (NPCs), even though TFEB itself is incapable of undergoing methylation. Specifically, protein phosphatase 1 catalytic subunit alpha (PPP1CA), newly identified to dephosphorylate TFEB, contains a K141 mono-methylated site. In degenerated NPCs, increased K141-methylation of PPP1CA disrupts its interaction with TEFB and subsequently blocks TEFB dephosphorylation and nuclear translocation, which eventually leads to autophagy deficiency and NPC senescence. In addition, we found that the PPP1CA-mediated targeting of TFEB is facilitated by the protein phosphatase 1 regulatory subunit 9B (PPP1R9B), which binds with PPP1CA and is also manipulated by K141 methylation. Further proteomic analysis revealed that the protein lysine methyltransferase suppressor of variegation 3-9 homologue 2 (SUV39H2) is responsible for the K141 mono-methylation of PPP1CA. Targeting SUV39H2 effectively mitigates NPC senescence and IDD progression, providing a potential therapeutic strategy for IDD intervention.
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Affiliation(s)
- Huaizhen Liang
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Rongjin Luo
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
- Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an, 710054, China
| | - Gaocai Li
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Weifeng Zhang
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Dingchao Zhu
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Di Wu
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Xingyu Zhou
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Bide Tong
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Bingjin Wang
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Xiaobo Feng
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Kun Wang
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China
| | - Yu Song
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
| | - Cao Yang
- Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
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Xu H, Li J, Fei Q, Jiang L. Contribution of immune cells to intervertebral disc degeneration and the potential of immunotherapy. Connect Tissue Res 2023; 64:413-427. [PMID: 37161923 DOI: 10.1080/03008207.2023.2212051] [Citation(s) in RCA: 11] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/27/2022] [Revised: 05/02/2023] [Accepted: 05/03/2023] [Indexed: 05/11/2023]
Abstract
Substantial evidence supports that chronic low back pain is associated with intervertebral disc degeneration (IDD), which is accompanied by decreased cell activity and matrix degradation. The role of immune cells, especially macrophages, in a variety of diseases has been extensively studied; therefore, their role in IDD has naturally attracted widespread scholarly interest. The IVD is considered to be an immunologically-privileged site given the presence of physical and biological barriers that include an avascular microenvironment, a high proteoglycan concentration, high physical pressure, the presence of apoptosis inducers such as Fas ligand, and the presence of notochordal cells. However, during IDD, immune cells with distinct characteristics appear in the IVD. Some of these immune cells release factors that promote the inflammatory response and angiogenesis in the disc and are, therefore, important drivers of IDD. Although some studies have elucidated the role of immune cells, no specific strategies related to systemic immunotherapy have been proposed. Herein, we summarize current knowledge of the presence and role of immune cells in IDD and consider that immunotherapy targeting immune cells may be a novel strategy for alleviating IDD symptoms.
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Affiliation(s)
- Hao Xu
- Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Juan Li
- Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Qinming Fei
- Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China
| | - Libo Jiang
- Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China
- Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Xiamen, Fujian Province, China
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Zhao WJ, Liu X, Hu M, Zhang Y, Shi PZ, Wang JW, Lu XH, Cheng XF, Tao YP, Feng XM, Wang YX, Zhang L. Quercetin ameliorates oxidative stress-induced senescence in rat nucleus pulposus-derived mesenchymal stem cells via the miR-34a-5p/SIRT1 axis. World J Stem Cells 2023; 15:842-865. [PMID: 37700818 PMCID: PMC10494568 DOI: 10.4252/wjsc.v15.i8.842] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/18/2023] [Revised: 05/25/2023] [Accepted: 06/27/2023] [Indexed: 08/25/2023] Open
Abstract
BACKGROUND Intervertebral disc degeneration (IDD) is a main contributor to low back pain. Oxidative stress, which is highly associated with the progression of IDD, increases senescence of nucleus pulposus-derived mesenchymal stem cells (NPMSCs) and weakens the differentiation ability of NPMSCs in degenerated intervertebral discs (IVDs). Quercetin (Que) has been demonstrated to reduce oxidative stress in diverse degenerative diseases. AIM To investigate the role of Que in oxidative stress-induced NPMSC damage and to elucidate the underlying mechanism. METHODS In vitro, NPMSCs were isolated from rat tails. Senescence-associated β-galactosidase (SA-β-Gal) staining, cell cycle, reactive oxygen species (ROS), real-time quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, and western blot analyses were used to evaluated the protective effects of Que. Meanwhile the relationship between miR-34a-5p and Sirtuins 1 (SIRT1) was evaluated by dual-luciferase reporter assay. To explore whether Que modulates tert-butyl hydroperoxide (TBHP)-induced senescence of NPMSCs via the miR-34a-5p/SIRT1 pathway, we used adenovirus vectors to overexpress and downregulate the expression of miR-34a-5p and used SIRT1 siRNA to knockdown SIRT1 expression. In vivo, a puncture-induced rat IDD model was constructed, and X rays and histological analysis were used to assess whether Que could alleviate IDD in vivo. RESULTS We found that TBHP can cause NPMSCs senescence changes, such as reduced cell proliferation ability, increased SA-β-Gal activity, cell cycle arrest, the accumulation of ROS, and increased expression of senescence-related proteins. While abovementioned senescence indicators were significantly alleviated by Que treatment. Que decreased the expression levels of senescence-related proteins (p16, p21, and p53) and senescence-associated secreted phenotype (SASP), including IL-1β, IL-6, and MMP-13, and it increased the expression of SIRT1. In addition, the protective effects of Que on cell senescence were partially reversed by miR-34a-5p overexpression and SIRT1 knockdown. In vivo, X-ray, and histological analyses indicated that Que alleviated IDD in a puncture-induced rat model. CONCLUSION In summary, the present study provides evidence that Que reduces oxidative stress-induced senescence of NPMSCs via the miR-34a/SIRT1 signaling pathway, suggesting that Que may be a potential agent for the treatment of IDD.
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Affiliation(s)
- Wen-Jie Zhao
- Graduate School, Dalian Medical University, Dalian 116044, Liaoning Province, China
| | - Xin Liu
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
| | - Man Hu
- Graduate School, Dalian Medical University, Dalian 116044, Liaoning Province, China
| | - Yu Zhang
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
| | - Peng-Zhi Shi
- Graduate School, Dalian Medical University, Dalian 116044, Liaoning Province, China
| | - Jun-Wu Wang
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
| | - Xu-Hua Lu
- Department of Orthopedics, Changzheng Hospital of The Second Military Medical University, Shanghai 200003, China
| | - Xiao-Fei Cheng
- Department of Orthopedic Surgery, Shanghai Key Laboratory of Orthopedics Implants, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
| | - Yu-Ping Tao
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
| | - Xin-Min Feng
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
| | - Yong-Xiang Wang
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China
| | - Liang Zhang
- Department of Orthopedics, Clinical Medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China.
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Liu C, Gao X, Lou J, Li H, Chen Y, Chen M, Zhang Y, Hu Z, Chang X, Luo M, Zhai Y, Li C. Aberrant mechanical loading induces annulus fibrosus cells apoptosis in intervertebral disc degeneration via mechanosensitive ion channel Piezo1. Arthritis Res Ther 2023; 25:117. [PMID: 37420255 PMCID: PMC10327399 DOI: 10.1186/s13075-023-03093-9] [Citation(s) in RCA: 9] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2022] [Accepted: 06/16/2023] [Indexed: 07/09/2023] Open
Abstract
BACKGROUND Intervertebral disc degeneration (IVDD) is closely associated with the structural damage in the annulus fibrosus (AF). Aberrant mechanical loading is an important inducement of annulus fibrosus cells (AFCs) apoptosis, which contributes to the AF structural damage and aggravates IVDD, but the underlying mechanism is still unclear. This study aims to investigate the mechanism of a mechanosensitive ion channel protein Piezo1 in aberrant mechanical loading-induced AFCs apoptosis and IVDD. METHODS Rats were subjected to lumbar instability surgery to induce the unbalanced dynamic and static forces to establish the lumbar instability model. MRI and histological staining were used to evaluate the IVDD degree. A cyclic mechanical stretch (CMS)-stimulated AFCs apoptosis model was established by a Flexcell system in vitro. Tunel staining, mitochondrial membrane potential (MMP) detection, and flow cytometry were used to evaluate the apoptosis level. The activation of Piezo1 was detected using western blot and calcium fluorescent probes. Chemical activator Yoda1, chemical inhibitor GSMTx4, and a lentiviral shRNA-Piezo1 system (Lv-Piezo1) were utilized to regulate the function of Piezo1. High-throughput RNA sequencing (RNA-seq) was used to explore the mechanism of Piezo1-induced AFCs apoptosis. The Calpain activity and the activation of Calpain2/Bax/Caspase3 axis were evaluated by the Calpain activity kit and western blot with the siRNA-mediated Calapin1 or Calpain2 knockdown. Intradiscal administration of Lv-Piezo1 was utilized to evaluate the therapeutic effect of Piezo1 silencing in IVDD rats. RESULTS Lumbar instability surgery promoted the expression of Piezo1 in AFCs and stimulated IVDD in rats 4 weeks after surgery. CMS elicited distinct apoptosis of AFCs, with enhanced Piezo1 activation. Yoda1 further promoted CMS-induced apoptosis of AFCs, while GSMTx4 and Lv-Piezo1 exhibited opposite effects. RNA-seq showed that knocking down Piezo1 inhibited the calcium signaling pathway. CMS enhanced Calpain activity and elevated the expression of BAX and cleaved-Caspase3. Calpain2, but not Calpain1 knockdown, inhibited the expression of BAX and cleaved-Caspase3 and alleviated AFCs apoptosis. Lv-Piezo1 significantly alleviated the progress of IVDD in rats after lumbar instability surgery. CONCLUSIONS Aberrant mechanical loading induces AFCs apoptosis to promote IVDD by activating Piezo1 and downstream Calpain2/BAX/Caspase3 pathway. Piezo1 is expected to be a potential therapeutic target in treating IVDD.
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Affiliation(s)
- Chenhao Liu
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
- Department of Orthopedics, Qinghai Provincial People's Hospital, Xining, 810007, Qinghai, China
| | - Xiaoxin Gao
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
| | - Jinhui Lou
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
| | - Haiyin Li
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
| | - Yuxuan Chen
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
- Center of Traumatic Orthopedics, People's Liberation Army 990 Hospital, Xinyang, 464000, Henan, China
| | - Molong Chen
- Department of Orthopedics/Sports Medicine Center, The First Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
| | - Yuyao Zhang
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
| | - Zhilei Hu
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
| | - Xian Chang
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China
| | - Menglin Luo
- Clinical Laboratory, Qinghai Provincial People's Hospital, Xining, 810007, Qinghai, China
| | - Yu Zhai
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China.
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China.
| | - Changqing Li
- Department of Orthopedics, The Second Affiliated Hospital of Army Medical University (The Third Military Medical University), Chongqing, 400038, China.
- State Key Laboratory of Trauma, Burns and Combined Injury, Chongqing, 400038, China.
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Chen T, Qian Q, Makvandi P, Zare EN, Chen Q, Chen L, Zhang Z, Zhou H, Zhou W, Wang H, Wang X, Chen Y, Zhou Y, Wu A. Engineered high-strength biohydrogel as a multifunctional platform to deliver nucleic acid for ameliorating intervertebral disc degeneration. Bioact Mater 2023; 25:107-121. [PMID: 37056255 PMCID: PMC10088054 DOI: 10.1016/j.bioactmat.2023.01.010] [Citation(s) in RCA: 12] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2022] [Revised: 01/07/2023] [Accepted: 01/12/2023] [Indexed: 02/04/2023] Open
Abstract
Intervertebral disc degeneration (IVDD) is a leading cause of low back pain. The strategy of using functional materials to deliver nucleic acids provides a powerful tool for ameliorating IVDD. However, the immunogenicity of nucleic acid vectors and the poor mechanical properties of functional materials greatly limit their effects. Herein, antagomir-204-3p (AM) shows low immunogenicity and effectively inhibits the apoptosis of nucleus pulposus cells. Moreover, a high-strength biohydrogel based on zinc-oxidized sodium alginate-gelatin (ZOG) is designed as a multifunctional nucleic acid delivery platform. ZOG loaded with AM (ZOGA) exhibits great hygroscopicity, antibacterial activity, biocompatibility, and biodegradability. Moreover, ZOGA can be cross-linked with nucleus pulposus tissue to form a high-strength collagen network that improves the mechanical properties of the intervertebral disc (IVD). In addition, ZOGA provides an advantageous microenvironment for genetic expression in which AM can play an efficient role in maintaining the metabolic balance of the extracellular matrix. The results of the radiological and histological analyses demonstrate that ZOGA restores the height of the IVD, retains moisture in the IVD, and maintains the tissue structure. The ZOGA platform shows the sustained release of nucleic acids and has the potential for application to ameliorate IVDD, opening a path for future studies related to IVD.
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Zhao X, Yuan J, Jia J, Zhang J, Liu J, Chen Q, Li T, Wu Z, Wu H, Miao X, Wu T, Li B, Cheng X. Role of non‑coding RNAs in cartilage endplate (Review). Exp Ther Med 2023; 26:312. [PMID: 37273754 PMCID: PMC10236100 DOI: 10.3892/etm.2023.12011] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/27/2022] [Accepted: 04/14/2023] [Indexed: 06/06/2023] Open
Abstract
Cartilage endplate (CEP) degeneration is considered one of the major causes of intervertebral disc degeneration (IDD), which causes non-specific neck and lower back pain. In addition, several non-coding RNAs (ncRNAs), including long ncRNAs, microRNAs and circular RNAs have been shown to be involved in the regulation of various diseases. However, the particular role of ncRNAs in CEP remains unclear. Identifying these ncRNAs and their interactions may prove to be is useful for the understanding of CEP health and disease. These RNA molecules regulate signaling pathways and biological processes that are critical for a healthy CEP. When dysregulated, they can contribute to the development disease. Herein, studies related to ncRNAs interactions and regulatory functions in CEP are reviewed. In addition, a summary of the current knowledge regarding the deregulation of ncRNAs in IDD in relation to their actions on CEP cell functions, including cell proliferation, apoptosis and extracellular matrix synthesis/degradation is presented. The present review provides novel insight into the pathogenesis of IDD and may shed light on future therapeutic approaches.
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Affiliation(s)
- Xiaokun Zhao
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Jinghong Yuan
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Jingyu Jia
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Jian Zhang
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Jiahao Liu
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Qi Chen
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Tao Li
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Zhiwen Wu
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Hui Wu
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Xinxin Miao
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Tianlong Wu
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
- Jiangxi Key Laboratory of Intervertebral Disc Disease, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Bin Li
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
- Jiangxi Key Laboratory of Intervertebral Disc Disease, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
| | - Xigao Cheng
- Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
- Jiangxi Key Laboratory of Intervertebral Disc Disease, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
- Institute of Minimally Invasive Orthopedics, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
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50
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Zhu G, Yang X, Zhou W, Lian X, Hao Y. PLAGL2 induces nucleus pulposus cell apoptosis via regulating RASSF5 expression and thus accelerates intervertebral disc degeneration. Exp Cell Res 2023:113699. [PMID: 37364764 DOI: 10.1016/j.yexcr.2023.113699] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2023] [Revised: 06/21/2023] [Accepted: 06/23/2023] [Indexed: 06/28/2023]
Abstract
Excessive apoptosis of nucleus pulposus (NP) cells is the main pathological change in intervertebral disc degeneration (IVDD) progression. Pleomorphic adenoma gene like-2 (PLAGL2) plays a key role in cell apoptosis, however, the effect of PLAGL2 on IVDD has not been clarified yet. In this study, we established mouse IVDD models via the annulus fibrosis needle puncture, TUNEL and safranin O staining were used to verify the successful establishment of IVDD models, and PLAGL2 expression was detected in disc tissues. Then, NP cells isolated from disc tissues were used to construct PLAGL2 knockdown cells. PLAGL2 expression in NP cells was analyzed with qRT-PCR and Western blot. The impact of PLAGL2 on the viability, apoptosis, and mitochondria function of NP cells was evaluated by MTT assay, TUNEL, JC1 staining, and flow cytometry assay. Additionally, the regulatory mechanism of PLAGL2 was further assessed. We found that PLAGL2 was upregulated in IVDD disc tissues and serum deprivation (SD)-stimulated NP cells. PLAGL2 knockdown inhibited apoptosis and mitochondria damage in NP cells. Moreover, knockdown of PLAGL2 downregulated the expression of downstream apoptosis-related factors RASSF5, Nip3, and p73. Mechanically, PLAGL2 transcriptionally activated RASSF5 via binding to its promoter. In general, our findings indicate that PLAGL2 induces apoptosis in NP cells and aggravates IVDD progression. This study provides a promising therapeutic target for IVDD treatment.
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Affiliation(s)
- GuangDuo Zhu
- Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
| | - XiaoWei Yang
- Department of Cardiovascular Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
| | - WeiWei Zhou
- Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
| | - Xu Lian
- Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
| | - YingJie Hao
- Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China.
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