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Tchoukalova YD, Shah MK, Myers CE, Zhang N, Lott DG. Optimization of seeding cell density for differentiation of adipose-derived stem cells into epithelial-like cells on bioengineered composite scaffolds. Differentiation 2025; 143:100870. [PMID: 40414118 DOI: 10.1016/j.diff.2025.100870] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/15/2024] [Revised: 05/15/2025] [Accepted: 05/16/2025] [Indexed: 05/27/2025]
Abstract
This study investigates the biological factors influencing the epithelial differentiation of adipose-derived stem cells (ASC) to develop an engineered upper airway construct. One fraction of ASC was seeded onto a fibrin sealant (Tisseel) matrix encompassing an additional equal fraction of ASC that has been integrated into a porous polyethylene scaffold (Medpor®). Constructs with ASC seeded at total densities of 5 × 105, 1 × 106, 2.5 × 106, and 5 × 106 cells cm-2 were cultured under submerged conditions for 11 days to achieve partial epithelial differentiation (PD). To simulate post-transplantation exposure to air and interaction with host epithelial cells, PD constructs with ASC at 5 × 106 cells cm-2 were transitioned to air-liquid interface (ALI) conditions for additional 10 days (PD/ALI-21d) or 21 days (PD/ALI-32d). The latter cultures were either maintained alone or co-cultured with bronchial epithelial cells (PD/ALI-32d + BEAS). Gene expressions of mesenchymal and epithelial basal, secretory, and ciliated cell markers were assessed and validated via immunohistochemistry. ASC seeded at 5 × 106 cells cm-2 achieved the highest partial epithelial differentiation, supporting the use of this density for further experiments. In PD/ALI-21d, basal and secretory epithelial marker gene expression significantly increased, while ciliated cell markers remained unchanged. In PD/ALI-32d, expression of basal and goblet cell markers and several mesenchymal stem cell markers decreased, but co-culturing with BEAS maintained the levels of their expression. These results indicate that long-term ALI cultures cannot sustain terminal differentiation of ASC into secretory phenotypes without co-culture with primary epithelial cells. In conclusion, partially differentiated ASC on constructs maintain a stem cell phenotype and may promote differentiation into basal/secretory phenotypes, but not ciliated cells. Enhancing ciliogenesis and ensuring ASC commitment to the epithelial lineage, require modifications to the study design.
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Affiliation(s)
- Yourka D Tchoukalova
- Mayo Clinic Arizona, Head and Neck Regenerative Medicine Laboratory, Scottsdale, AZ, USA
| | - Manisha K Shah
- Mayo Clinic Arizona, Head and Neck Regenerative Medicine Laboratory, Scottsdale, AZ, USA
| | - Cheryl E Myers
- Mayo Clinic Arizona, Head and Neck Regenerative Medicine Laboratory, Scottsdale, AZ, USA
| | - Nan Zhang
- Mayo Clinic Arizona, Department of Quantitative Health Science Research, Scottsdale, AZ, USA
| | - David G Lott
- Mayo Clinic Arizona, Head and Neck Regenerative Medicine Laboratory, Scottsdale, AZ, USA; Mayo Clinic Arizona, Department of Otolaryngology - Head and Neck Surgery, Division of Laryngology, Phoenix, AZ, USA.
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Hazrati R, Alizadeh E, Soltani S, Keyhanvar P, Davaran S. Development of a Composite Hydrogel Containing Statistically Optimized PDGF-Loaded Polymeric Nanospheres for Skin Regeneration: In Vitro Evaluation and Stem Cell Differentiation Studies. ACS OMEGA 2024; 9:15114-15133. [PMID: 38585049 PMCID: PMC10993260 DOI: 10.1021/acsomega.3c09391] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/24/2023] [Revised: 02/21/2024] [Accepted: 03/05/2024] [Indexed: 04/09/2024]
Abstract
Platelet-derived growth factor-BB (PDGF-BB) is a polypeptide growth factor generated by platelet granules faced to cytokines. It plays a role in forming and remodeling various tissue types, including epithelial tissue, through interaction with cell-surface receptors on most mesenchymal origin cells. However, it breaks down quickly in biological fluids, emphasizing the importance of preserving them from biodegradation. To address this challenge, we formulated and evaluated PDGF-encapsulated nanospheres (PD@PCEC) using polycaprolactone-polyethylene glycol-polycaprolactone. PD@PCECs were fabricated through the triple emulsion methodology and optimized by using the Box-Behnken design. The encapsulation efficiency (EE) of nanoencapsulated PDGF-BB was investigated concerning four variables: stirring rate (X1), stirring duration (X2), poly(vinyl alcohol) concentration (X3), and PDGF-BB concentration (X4). The selected optimized nanospheres were integrated into a gelatin-collagen scaffold (PD@PCEC@GC) and assessed for morphology, biocompatibility, in vitro release, and differentiation-inducing activity in human adipose-derived stem cells (hADSCs). The optimized PD@PCEC nanospheres exhibited a particle size of 177.9 ± 91 nm, a zeta potential of 5.2 mV, and an EE of 87.7 ± 0.44%. The release profile demonstrated approximately 85% of loaded PDGF-BB released during the first 360 h, with a sustained release over the entire 504 h period, maintaining bioactivity of 87.3%. The study also included an evaluation of the physicochemical properties of the scaffolds and an assessment of hADSC adhesion to the scaffold's surface. Additionally, hADSCs cultivated within the scaffold effectively differentiated into keratinocyte-like cells (KLCs) over 21 days, evidenced by morphological changes and upregulation of keratinocyte-specific genes, including cytokeratin 18, cytokeratin 19, and involucrin, at both transcriptional and protein levels.
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Affiliation(s)
- Raheleh Hazrati
- Department
of Medicinal Chemistry, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz 51664-14766, Iran
- Research
Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz 51664, Iran
| | - Effat Alizadeh
- Department
of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz 51664-15731, Iran
| | - Somaieh Soltani
- Department
of Medicinal Chemistry, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz 51664-14766, Iran
| | - Peyman Keyhanvar
- Department
of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz 51664-15731, Iran
| | - Soodabeh Davaran
- Department
of Medicinal Chemistry, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz 51664-14766, Iran
- Research
Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz 51664, Iran
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Hazrati R, Davaran S, Keyhanvar P, Soltani S, Alizadeh E. A Systematic Review of Stem Cell Differentiation into Keratinocytes for Regenerative Applications. Stem Cell Rev Rep 2024; 20:362-393. [PMID: 37922106 DOI: 10.1007/s12015-023-10636-9] [Citation(s) in RCA: 2] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 09/25/2023] [Indexed: 11/05/2023]
Abstract
To improve wound healing or treatment of other skin diseases, and provide model cells for skin biology studies, in vitro differentiation of stem cells into keratinocyte-like cells (KLCs) is very desirable in regenerative medicine. This study examined the most recent advancements in in vitro differentiation of stem cells into KLCs, the effect of biofactors, procedures, and preparation for upcoming clinical cases. A range of stem cells with different origins could be differentiated into KLCs under appropriate conditions. The most effective ways of stem cell differentiation into keratinocytes were found to include the co-culture with primary epithelial cells and keratinocytes, and a cocktail of growth factors, cytokines, and small molecules. KLCs should also be supported by biomaterials for the extracellular matrix (ECM), which replicate the composition and functionality of the in vivo extracellular matrix (ECM) and, thus, support their phenotypic and functional characteristics. The detailed efficient characterization of different factors, and their combinations, could make it possible to find the significant inducers for stem cell differentiation into epidermal lineage. Moreover, it allows the development of chemically known media for directing multi-step differentiation procedures.In conclusion, the differentiation of stem cells to KLCs is feasible and KLCs were used in experimental, preclinical, and clinical trials. However, the translation of KLCs from in vitro investigational system to clinically valuable cells is challenging and extremely slow.
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Affiliation(s)
- Raheleh Hazrati
- Department of Medicinal Chemistry, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
- Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran
| | - Soodabeh Davaran
- Department of Medicinal Chemistry, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
- Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran.
| | - Peyman Keyhanvar
- Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran
| | - Somaieh Soltani
- Department of Medicinal Chemistry, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
| | - Effat Alizadeh
- Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
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Habib R, Fahim S, Wahid M, Ainuddin J. Optimisation of a Method for the Differentiation of Human Umbilical Cord-derived Mesenchymal Stem Cells Toward Renal Epithelial-like Cells. Altern Lab Anim 2023; 51:363-375. [PMID: 37831588 DOI: 10.1177/02611929231207774] [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: 10/15/2023]
Abstract
Human umbilical cord-derived mesenchymal stem cells (hucMSCs) can differentiate into multiple cell lineages, but few methods have been developed to generate kidney lineage cells. Due to their human origin, pluripotent nature and immunomodulatory properties, these stem cells are attractive candidates for clinical applications such as the repair or regeneration of damaged organs. This study evaluated the renal differentiation potential of hucMSCs, when exposed for 10 days to optimised concentrations of retinoic acid, activin-A and bone morphogenetic protein-7 (BMP-7) in various combinations, with and without the priming of the cells with a Wnt signalling pathway activator (CHIR99021). The hucMSCs were isolated and characterised according to surface marker expression (CD73, CD90, CD44, CD146 and CD8) and tri-lineage differentiation potential. The expression of key marker genes (OSR1, TBXT, HOXA13, SIX2, PAX2, KRT18 and ZO1) was examined by qRT-PCR. Specific marker protein expression (E-cadherin, cytokeratin-8 and cytokeratin-19) was analysed by immunocytochemistry. CHIR99021-primed cells treated with the retinoic acid, activin-A and BMP-7 cocktail showed epithelial cell-like differentiation - i.e. distinct phenotypic changes, as well as upregulated gene and protein expression, were observed that were consistent with an epithelial cell phenotype. Thus, our results showed that hucMSCs can efficiently differentiate into renal epithelial-like cells. This work may help in the development of focused therapeutic strategies, in which lineage-defined human stem cells can be used for renal regeneration.
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Affiliation(s)
- Rakhshinda Habib
- Dow Research Institute of Biotechnology and Biomedical Sciences, Dow University of Health Sciences (Ojha campus), Karachi, Pakistan
| | - Shumaila Fahim
- Dow Research Institute of Biotechnology and Biomedical Sciences, Dow University of Health Sciences (Ojha campus), Karachi, Pakistan
| | - Mohsin Wahid
- Department of Pathology, Dow International Medical College, Dow University of Health Sciences (Ojha campus), Karachi, Pakistan
| | - Jahanara Ainuddin
- Department of Gynaecology and Obstetrics, Dow University Hospital, Karachi, Pakistan
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Chen J, Ye P, Gu R, Zhu H, He W, Mu X, Wu X, Pang H, Han F, Nie X. Neuropeptide substance P: A promising regulator of wound healing in diabetic foot ulcers. Biochem Pharmacol 2023; 215:115736. [PMID: 37549795 DOI: 10.1016/j.bcp.2023.115736] [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: 06/12/2023] [Revised: 08/01/2023] [Accepted: 08/02/2023] [Indexed: 08/09/2023]
Abstract
In the past, neuropeptide substance P (SP) was predominantly recognized as a neuroinflammatory factor, while its potent healing activity was overlooked. This paper aims to review the regulatory characteristics of neuropeptide SP in both normal and diabetic wound healing. SP actively in the regulation of wound healing-related cells directly and indirectly, exhibiting robust inflammatory properties, promoting cell proliferation and migration and restoring the activity and paracrine ability of skin cells under diabetic conditions. Furthermore, SP not only regulates healing-related cells but also orchestrates the immune environment, thereby presenting unique and promising application prospects in wound intervention. As new SP-based preparations are being explored, SP-related drugs are poised to become an effective therapeutic intervention for diabetic foot ulcers (DFU).
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Affiliation(s)
- Jitao Chen
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China
| | - Penghui Ye
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China
| | - Rifang Gu
- University Medical Office, Zunyi Medical University, Zunyi 563000, China
| | - Huan Zhu
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China
| | - Wenjie He
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China
| | - Xingrui Mu
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China
| | - Xingqian Wu
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China
| | - Huiwen Pang
- Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, Australia
| | - Felicity Han
- Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, Australia
| | - Xuqiang Nie
- College of Pharmacy, Zunyi Medical University, Zunyi 563000, China; Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, Australia; Key Lab of the Basic Pharmacology of the Ministry of Education & Joint International Research Laboratory of Ethnomedicine of Chinese Ministry of Education, Zunyi Medical University, Zunyi 563000, China.
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Wang Z, Lai J, Li Y, Zhou H, Alhaskawi A, Li P, Shen X, Lu H, Tu T. Could E-cadherin overexpression promote epithelial differentiation of human adipose-derived stem cells by mediating mesenchymal-to-epithelial transition? Med Hypotheses 2023; 171:111016. [DOI: 10.1016/j.mehy.2023.111016] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023]
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ÖZGENÇ Ö, ÖZEN A. Osteogenic Differentiation of Canine Adipose Derived Mesenchymal Stem Cells on B-TCP and B-TCP/Collagen Biomaterials. ANKARA ÜNIVERSITESI VETERINER FAKÜLTESI DERGISI 2022. [DOI: 10.33988/auvfd.1130705] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/09/2023]
Abstract
Mesenchymal stem cells are adult stem cells that have the ability to differentiate into osteogenic, chondrogenic, adipogenic and myogenic lineages. In the field of orthopedics and traumatology, mesenchymal stem cells in combination with biomaterials are used especially for the treatment of bone fractures and diseases in both humans and animals. The purpose of this study is to promote growth, proliferation and osteogenic differentiation of mesenchymal stem cells that were isolated from the adipose tissue of canines on B-TCP (Beta-tricalcium phosphate) and B-TCP/Collagen biomaterials. MTT analysis was performed to test the cell adhesion and proliferation on B-TCP and B-TCP/Collagen biomaterials that were used to mimic the extracellular matrix of three-dimensional bone tissue. Scanning electron microscope analysis was performed to show general surface characters of B-TCP and B-TCP /Collagen biomaterials. The osteoinductive capacities of the B-TCP and B-TCP/Collagen biomaterials were determined by alkaline phosphatase and Von Kossa stainings, and RT-PCR analysis. The ALP activity of the B-TCP/Col containing material was significantly higher than the B-TCP on the first days. In terms of gene expression, there were no significant differences except 14th-day SPARC gene expression. The results of Von Kossa staining indicate that B-TCP/Col has above the desired level degradation capacity. As a result of this research, although it is advantageous in terms of alkaline phosphatase activity and osteogenic gene expression compared to B-TCP material, it is thought that B-TCP/Collagen biomaterial should be developed for use in bone tissue engineering due to its high degradation property.
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Liu L, Wang J, Li Y, Liu B, Zhang W, An W, Wang Q, Xu B, Zhao L, Ma C. Laminin 332-functionalized coating to regulate the behavior of keratinocytes and gingival mesenchymal stem cells to enhance implant soft tissue sealing. Regen Biomater 2022; 9:rbac054. [PMID: 36072266 PMCID: PMC9438747 DOI: 10.1093/rb/rbac054] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/02/2022] [Revised: 06/21/2022] [Accepted: 07/10/2022] [Indexed: 11/14/2022] Open
Abstract
Peri-implant epithelial sealing is the first line of defense against external pathogens or stimuli; hence, an essential process to prevent peri-implantitis. Laminin 332 (LN332) is the main component of the internal basal lamina and participates in peri-implant epithelial sealing by forming hemidesmosomes (HDs) with integrin α6β4. In this work, poly (D, L-lactide) (PDLLA)-LN332 composite coating was successfully constructed by a method similar to layer-by-layer assembly, displaying staged LN332 release for as long as 28 days. The PDLLA-LN332 composite coating can activate the intracellular PI3K-Akt pathway via binding to cellular integrin α6β4, which can promote adhesion, migration and proliferation of HaCaT cells and further enhance the expression of keratinocyte HD-related molecules, including integrin α6β4, LN332 and plectin. Furthermore, the PDLLA-LN332 composite coating can promote the adhesion, spreading and proliferation of gingival mesenchymal stem cells and accelerate their epithelial differentiation. Therefore, the PDLLA-LN332 composite coating can enhance implant soft tissue sealing, warranting further in vivo study.
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Affiliation(s)
- Lipeng Liu
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
| | - Jing Wang
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
| | - Ying Li
- The Fourth Military Medical University Department of Stomatology, Air Force Medical Center, , Beijing, 100142, China
| | - Bing Liu
- The Fourth Military Medical University Department of Stomatology, Air Force Medical Center, , Beijing, 100142, China
| | - Wei Zhang
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
| | - Weikang An
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
| | - Qing Wang
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
| | - Boya Xu
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
| | - Lingzhou Zhao
- The Fourth Military Medical University Department of Stomatology, Air Force Medical Center, , Beijing, 100142, China
| | - Chufan Ma
- The Fourth Military Medical University State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Key Laboratory of Stomatology, Department of Prosthodontics, School of Stomatology, , Xi'an, Shaanxi, 710032, China
- The Fourth Military Medical University Department of Stomatology, Air Force Medical Center, , Beijing, 100142, China
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Effects of Hypoxia on RNA Cargo in Extracellular Vesicles from Human Adipose-Derived Stromal/Stem Cells. Int J Mol Sci 2022; 23:ijms23137384. [PMID: 35806391 PMCID: PMC9266528 DOI: 10.3390/ijms23137384] [Citation(s) in RCA: 6] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/16/2022] [Revised: 06/28/2022] [Accepted: 06/30/2022] [Indexed: 02/01/2023] Open
Abstract
Mesenchymal stromal/stem cells and their derivates are the most promising cell source for cell therapies in regenerative medicine. The application of extracellular vesicles (EVs) as cell-free therapeuticals requires particles with a maximum regenerative capability to enhance tissue and organ regeneration. The cargo of mRNA and microRNA (miR) in EVs after hypoxic preconditioning has not been extensively investigated. Therefore, the aim of our study was the characterization of mRNA and the miR loading of EVs. We further investigated the effects of the isolated EVs on renal tubular epithelial cells in vitro. We found 3131 transcripts to be significantly regulated upon hypoxia. Only 15 of these were downregulated, but 3116 were up-regulated. In addition, we found 190 small RNAs, 169 of these were miRs and 21 were piwi-interacting RNAs (piR). However, only 18 of the small RNAs were significantly altered, seven were miRs and 11 were piRs. Interestingly, all seven miRs were down-regulated after hypoxic pretreatment, whereas all 11 piRs were up-regulated. Gene ontology term enrichment and miR-target enrichment analysis of the mRNAs and miR were also performed in order to study the biological background. Finally, the therapeutic effect of EVs on human renal tubular epithelial cells was shown by the increased expression of three anti-inflammatory molecules after incubation with EVs from hypoxic pretreatment. In summary, our study demonstrates the altered mRNA and miR load in EVs after hypoxic preconditioning, and their anti-inflammatory effect on epithelial cells.
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Lott DG, Shah M, Myers C, McPhail M, Neubauer J, Struthers J, Madsen CS, Grandjean D, Zacharias SRC, Tchoukalova YD. A Tissue Engineered Construct for Laryngeal Regeneration: A Proof-of-Concept Device Design Study. Laryngoscope 2022; 132 Suppl 9:S1-S11. [PMID: 35084750 DOI: 10.1002/lary.30029] [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: 08/30/2021] [Revised: 12/21/2021] [Accepted: 01/07/2022] [Indexed: 11/06/2022]
Abstract
OBJECTIVES/HYPOTHESIS Develop a patient-specific tissue engineered construct for laryngeal reconstruction following a partial laryngectomy. STUDY DESIGN Bench and animal research. METHODS A construct made from a porous polyethylene scaffold shaped in a canine-specific configuration and seeded with autologous canine adipose-derived stem cells in fibrin glue was implanted in a canine following a partial laryngectomy. After 1 year, the construct was first evaluated in vivo with high-speed imaging and acoustic-aerodynamic measures. It was then explanted and evaluated histologically. RESULTS The canine study at 1 year revealed the construct provided voicing (barking) with acoustic and aerodynamic measures within normal ranges. The canine was able to eat and breathe normally without long-term support. The construct was integrated with epithelialization of all areas except the medial portion of the vocal fold structure. No anti-infective agents were needed after the standard perioperative medications were completed. CONCLUSION This study provided a successful first step toward developing a patient-specific composite construct for patients undergoing partial laryngectomies. LEVEL OF EVIDENCE Not Applicable Laryngoscope, 2022.
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Affiliation(s)
- David G Lott
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A.,Division of Laryngology, Department of Otolaryngology-Head and Neck Surgery, Mayo Clinic Arizona, Phoenix, Arizona, U.S.A
| | - Manisha Shah
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
| | - Cheryl Myers
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
| | - Michael McPhail
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
| | - Juergen Neubauer
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
| | - Jason Struthers
- Department of Pathology and Population Medicine of Midwestern University's College of Veterinary Medicine, Glendale, Arizona, U.S.A
| | - Cathy S Madsen
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
| | - Danielle Grandjean
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
| | - Stephanie R C Zacharias
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A.,Division of Laryngology, Department of Otolaryngology-Head and Neck Surgery, Mayo Clinic Arizona, Phoenix, Arizona, U.S.A
| | - Yourka D Tchoukalova
- Head and Neck Regenerative Medicine Laboratory, Mayo Clinic Arizona, Scottsdale, Arizona, U.S.A
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HEK293-Conditioned Medium Altered the Expression of Renal Markers WT1, CD2AP, and CDH16 in the Human Adipose Mesenchymal Stem Cells. REGENERATIVE ENGINEERING AND TRANSLATIONAL MEDICINE 2022. [DOI: 10.1007/s40883-021-00246-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
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12
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Tripathy S, VinayKumar D, Mohsina S, Sharma R, Bhatia A. Histological analysis of the effect of nanofat grafting in scar rejuvenation. J Cutan Aesthet Surg 2022; 15:147-153. [PMID: 35965912 PMCID: PMC9364463 DOI: 10.4103/jcas.jcas_106_21] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/04/2022] Open
Abstract
Introduction: Aims: Materials and Methods: Results: Conclusion:
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Angiogenic Effects and Crosstalk of Adipose-Derived Mesenchymal Stem/Stromal Cells and Their Extracellular Vesicles with Endothelial Cells. Int J Mol Sci 2021; 22:ijms221910890. [PMID: 34639228 PMCID: PMC8509224 DOI: 10.3390/ijms221910890] [Citation(s) in RCA: 52] [Impact Index Per Article: 13.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/16/2021] [Revised: 10/02/2021] [Accepted: 10/04/2021] [Indexed: 12/13/2022] Open
Abstract
Adipose-derived mesenchymal stem/stromal cells (ASCs) are an adult stem cell population able to self-renew and differentiate into numerous cell lineages. ASCs provide a promising future for therapeutic angiogenesis due to their ability to promote blood vessel formation. Specifically, their ability to differentiate into endothelial cells (ECs) and pericyte-like cells and to secrete angiogenesis-promoting growth factors and extracellular vesicles (EVs) makes them an ideal option in cell therapy and in regenerative medicine in conditions including tissue ischemia. In recent angiogenesis research, ASCs have often been co-cultured with an endothelial cell (EC) type in order to form mature vessel-like networks in specific culture conditions. In this review, we introduce co-culture systems and co-transplantation studies between ASCs and ECs. In co-cultures, the cells communicate via direct cell-cell contact or via paracrine signaling. Most often, ASCs are found in the perivascular niche lining the vessels, where they stabilize the vascular structures and express common pericyte surface proteins. In co-cultures, ASCs modulate endothelial cells and induce angiogenesis by promoting tube formation, partly via secretion of EVs. In vivo co-transplantation of ASCs and ECs showed improved formation of functional vessels over a single cell type transplantation. Adipose tissue as a cell source for both mesenchymal stem cells and ECs for co-transplantation serves as a prominent option for therapeutic angiogenesis and blood perfusion in vivo.
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Tissue engineering: recent advances and review of clinical outcome for urethral strictures. Curr Opin Urol 2021; 31:498-503. [PMID: 34397507 DOI: 10.1097/mou.0000000000000921] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023]
Abstract
PURPOSE OF REVIEW Urethrotomy remains the first-line therapy in the treatment of a urethral stricture despite data showing no real chance of a cure after repeated urethrotomies. An anastomotic or an augmentation urethroplasty using oral mucosa can be offered to patients following failed urethrotomy. The potential for a tissue engineered solution as an alternative to native tissue has been explored in recent years and is reviewed in this article. RECENT FINDINGS More than 80 preclinical studies have investigated a tissue-engineered approach for urethral reconstruction mostly using decellularized natural scaffolds derived from natural extracellular matrix with or without cell seeding. The animal models used in preclinical testing are not representative of disease processes seen with strictures in man. The available clinical studies are based on small noncontrolled series. SUMMARY There is a potential role for tissue engineering to provide a material for substitution urethroplasty and work has demonstrated this. Further work will require a rigorous basic science programme and adequate evaluation of the material prior to its introduction into clinical practice. The research with tissue engineering applied to the urethra has not yet been resulted in a widely available material for clinical use that approaches the efficacy seen with the use of autologous grafts.
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Geßner A, Koch B, Klann K, Fuhrmann DC, Farmand S, Schubert R, Münch C, Geiger H, Baer PC. Characterization of Extracellular Vesicles from Preconditioned Human Adipose-Derived Stromal/Stem Cells. Int J Mol Sci 2021; 22:ijms22062873. [PMID: 33808970 PMCID: PMC7999156 DOI: 10.3390/ijms22062873] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2021] [Revised: 03/09/2021] [Accepted: 03/10/2021] [Indexed: 12/14/2022] Open
Abstract
Cell-free therapy using extracellular vesicles (EVs) from adipose-derived mesenchymal stromal/stem cells (ASCs) seems to be a safe and effective therapeutic option to support tissue and organ regeneration. The application of EVs requires particles with a maximum regenerative capability and hypoxic culture conditions as an in vitro preconditioning regimen has been shown to alter the molecular composition of released EVs. Nevertheless, the EV cargo after hypoxic preconditioning has not yet been comprehensively examined. The aim of the present study was the characterization of EVs from hypoxic preconditioned ASCs. We investigated the EV proteome and their effects on renal tubular epithelial cells in vitro. While no effect of hypoxia was observed on the number of released EVs and their protein content, the cargo of the proteins was altered. Proteomic analysis showed 41 increased or decreased proteins, 11 in a statistically significant manner. Furthermore, the uptake of EVs in epithelial cells and a positive effect on oxidative stress in vitro were observed. In conclusion, culture of ASCs under hypoxic conditions was demonstrated to be a promising in vitro preconditioning regimen, which alters the protein cargo and increases the anti-oxidative potential of EVs. These properties may provide new potential therapeutic options for regenerative medicine.
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Affiliation(s)
- Alec Geßner
- Division of Nephrology, Department of Internal Medicine III, University Hospital, Goethe-University, 60596 Frankfurt/M., Germany; (A.G.); (B.K.); (S.F.); (H.G.)
| | - Benjamin Koch
- Division of Nephrology, Department of Internal Medicine III, University Hospital, Goethe-University, 60596 Frankfurt/M., Germany; (A.G.); (B.K.); (S.F.); (H.G.)
| | - Kevin Klann
- Institute of Biochemistry II, Faculty of Medicine, Goethe-University, 60596 Frankfurt/M., Germany; (K.K.); (C.M.)
| | - Dominik C. Fuhrmann
- Institute of Biochemistry I, Faculty of Medicine, Goethe-University Frankfurt, 60590 Frankfurt, Germany;
| | - Samira Farmand
- Division of Nephrology, Department of Internal Medicine III, University Hospital, Goethe-University, 60596 Frankfurt/M., Germany; (A.G.); (B.K.); (S.F.); (H.G.)
| | - Ralf Schubert
- Division of Allergology, Pneumology and Cystic Fibrosis, Department for Children and Adolescents, University Hospital, Goethe-University, 60596 Frankfurt/M., Germany;
| | - Christian Münch
- Institute of Biochemistry II, Faculty of Medicine, Goethe-University, 60596 Frankfurt/M., Germany; (K.K.); (C.M.)
| | - Helmut Geiger
- Division of Nephrology, Department of Internal Medicine III, University Hospital, Goethe-University, 60596 Frankfurt/M., Germany; (A.G.); (B.K.); (S.F.); (H.G.)
| | - Patrick C. Baer
- Division of Nephrology, Department of Internal Medicine III, University Hospital, Goethe-University, 60596 Frankfurt/M., Germany; (A.G.); (B.K.); (S.F.); (H.G.)
- Correspondence: ; Tel.: +49-6301-5554
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Al-Ghadban S, Bunnell BA. Adipose Tissue-Derived Stem Cells: Immunomodulatory Effects and Therapeutic Potential. Physiology (Bethesda) 2021; 35:125-133. [PMID: 32027561 DOI: 10.1152/physiol.00021.2019] [Citation(s) in RCA: 82] [Impact Index Per Article: 20.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022] Open
Abstract
Adipose-derived stem cells (ASCs) can self-renew and differentiate along multiple cell lineages. ASCs are also potently anti-inflammatory due to their inherent ability to regulate the immune system by secreting anti-inflammatory cytokines and growth factors that play a crucial role in the pathology of many diseases, including multiple sclerosis, diabetes mellitus, Crohn's, SLE, and graft-versus-host disease. The immunomodulatory effects and mechanisms of action of ASCs on pathological conditions are reviewed here.
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Affiliation(s)
- Sara Al-Ghadban
- Center for Stem Cell Research & Regenerative Medicine, Tulane University School of Medicine, New Orleans, Louisiana
| | - Bruce A Bunnell
- Center for Stem Cell Research & Regenerative Medicine, Tulane University School of Medicine, New Orleans, Louisiana.,Department of Pharmacology, School of Medicine, Tulane University, New Orleans, Louisiana
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17
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Tatebayashi R, Nakamura S, Minabe S, Furusawa T, Abe R, Kajisa M, Morita Y, Ohkura S, Kimura K, Matsuyama S. Gene-expression profile and postpartum transition of bovine endometrial side population cells†. Biol Reprod 2021; 104:850-860. [PMID: 33438005 DOI: 10.1093/biolre/ioab004] [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: 03/09/2020] [Revised: 12/11/2020] [Accepted: 01/07/2021] [Indexed: 11/12/2022] Open
Abstract
The mechanism of bovine endometrial regeneration after parturition remains unclear. Here, we hypothesized that bovine endometrial stem/progenitor cells participate in the postpartum regeneration of the endometrium. Flow cytometry analysis identified the presence of side population (SP) cells among endometrial stromal cells. Endometrial SP cells were shown to differentiate into osteoblasts and adipocytes. RNA-seq data showed that the gene expression pattern was different between bovine endometrial SP cells and main population cells. Gene Set Enrichment Analysis identified the enrichment of stemness genes in SP cells. Significantly (false discovery rate < 0.01) upregulated genes in SP cells contained several stem cell marker genes. Gene ontology (GO) analysis of the upregulated genes in SP cells showed enrichment of terms related to RNA metabolic process and transcription. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of upregulated genes in SP cells revealed enrichment of signaling pathways associated with maintenance and differentiation of stem/progenitor cells. The terms involved in TCA cycles were enriched in GO and KEGG pathway analysis of downregulated genes in SP cells. These results support the assumption that bovine endometrial SP cells exhibit characteristics of somatic stem/progenitor cells. The ratio of SP cells to endometrial cells was lowest on days 9-11 after parturition, which gradually increased thereafter. SP cells were shown to differentiate into epithelial cells. Collectively, these results suggest that bovine endometrial SP cells were temporarily reduced immediately after calving possibly due to their differentiation to provide new endometrial cells.
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Affiliation(s)
- Ryoki Tatebayashi
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan
| | - Sho Nakamura
- Faculty of Veterinary Medicine, Okayama University of Science, Ehime, Japan
| | - Shiori Minabe
- Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, Tokyo, Japan
| | - Tadashi Furusawa
- Division of Animal Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Ibaraki, Japan
| | - Ryoya Abe
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan
| | - Miki Kajisa
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan
| | - Yasuhiro Morita
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan.,Asian Satellite Campuses Institute, Nagoya University, Nagoya, Japan
| | - Satoshi Ohkura
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan
| | - Koji Kimura
- Laboratory of Reproductive Physiology, Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan
| | - Shuichi Matsuyama
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Aichi, Japan
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18
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Smolar J, Horst M, Salemi S, Eberli D. Predifferentiated Smooth Muscle-Like Adipose-Derived Stem Cells for Bladder Engineering. Tissue Eng Part A 2020; 26:979-992. [DOI: 10.1089/ten.tea.2019.0216] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022] Open
Affiliation(s)
- Jakub Smolar
- Department of Urology, University Hospital Zurich, Zurich, Switzerland
| | - Maya Horst
- Division of Pediatric Urology, University Children's Hospital Zurich, Zurich, Switzerland
| | - Souzan Salemi
- Department of Urology, University Hospital Zurich, Zurich, Switzerland
| | - Daniel Eberli
- Department of Urology, University Hospital Zurich, Zurich, Switzerland
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19
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Klinger M, Marazzi M, Vigo D, Torre M. Fat Injection for Cases of Severe Burn Outcomes: A New Perspective of Scar Remodeling and Reduction. Aesthetic Plast Surg 2020; 44:1278-1282. [PMID: 32844266 DOI: 10.1007/s00266-020-01813-z] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
BACKGROUND Despite civilization and progress, burns occur frequently in the world. Remarkable discoveries of wound healing mechanisms have been reported. On the other hand, long-term outcomes from burn injuries represent a barrier to improvement of patients' social, functional, and psychological condition. Lipofilling, described since the 1980s, currently is used for several clinical applications. This study aimed to verify whether lipofilling could ameliorate scar remodeling in three clinical cases. METHODS Three adult patients with hemifacial hypertrophic scars and keloids resulting from severe burns 2 to 13 years previously were selected. The patients were treated by injection of adipose tissue harvested from abdominal subcutaneous fat and processed according to Coleman's technique. Two injections (with a 13-month interval between) were administered at the dermohypodermal junction. Histologic examination of scar tissue punch biopsies (hematoxylin-eosin staining) before and after the treatment was performed as well as magnetic resonance scan with contrast. RESULTS The clinical appearance and subjective patient feelings after a 6-month follow-up period suggest considerable improvement in the mimic features, skin texture, and thickness. Histologic examination shows patterns of new collagen deposition, local hypervascularity, and dermal hyperplasia in the context of new tissue, with high correspondence to the original. CONCLUSIONS The preliminary results show that lipofilling improves scar quality and suggest a tissue regeneration enhancing process.
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Affiliation(s)
- M Klinger
- Università degli Studi di Milano, Istituto di Chirurgia Plastica, Unità Operativa di Chirurgia Plastica, IRCCS Istituto Clinico Humanitas, Via Manzoni 56, 20089, Rozzano, Italy.
| | - M Marazzi
- Centro di Riferimento Regionale Colture Cellulari, Ospedale Niguarda ''Ca' Granda'', Milano, Italy
| | - D Vigo
- Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare, Università degli Studi di Milano, Milano, Italy
| | - M Torre
- Dipartimento di Chimica Farmaceutica, Università degli Studi di Pavia, Pavia, Italy
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20
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Shi R, Lian W, Jin Y, Cao C, Han S, Yang X, Zhao S, Li M, Zhao H. Role and effect of vein-transplanted human umbilical cord mesenchymal stem cells in the repair of diabetic foot ulcers in rats. Acta Biochim Biophys Sin (Shanghai) 2020; 52:620-630. [PMID: 32484226 PMCID: PMC7333920 DOI: 10.1093/abbs/gmaa039] [Citation(s) in RCA: 25] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2019] [Revised: 01/15/2020] [Accepted: 03/31/2020] [Indexed: 12/13/2022] Open
Abstract
Diabetic foot ulcer (DFU) is one of diabetic complications, which is frequently present and tormented in diabetes mellitus. Most multipotent mesenchymal stromal cells (MSCs) are capable of immune evasion, providing an allogeneic, ready-to-use, cell product option for therapeutic applications. The beneficial effect of MSCs for the treatment of a variety of traumatic injuries, such as open wounds, has been extensively explored. In this study, a rat DFU model was used to simulate the pathophysiology of clinical patients and to investigate the localization of human umbilical cord mesenchymal stem cells (hUC-MSCs) after intravenous transplantation and its role in DFU healing, so as to evaluate the potential of hUC-MSCs in the treatment of DFU. The diabetic rat model was established by streptozotocin injection, which was used to create full-thickness foot dorsal skin wounds to mimic DFU by a 6-mm skin biopsy punch and a Westcott scissor. The hUC-MSCs were transplanted through femoral vein, and the ulcer cicatrization situation and the fate of hUC-MSCs were evaluated. Our data suggest that intravenously transplantated hUC-MSCs have the ability to migrate and locate to the wound tissue and are helpful to wound healing in DFU rats, partly by regulating inflammation, trans-differentiation and providing growth factors that promote angiogenesis, cell proliferation and collagen deposition. Herein, we demonstrate that hUC-MSC transplantation is able to accelerate DFU healing in rats and transplantation of exogenous stem cells may be a potential strategy for clinical application in DFUs.
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Affiliation(s)
- Rongfeng Shi
- Department of Interventional Radiology, Affiliated Hospital of Nantong University, Nantong 226001, China
| | - Weishuai Lian
- Department of Interventional & Vascular Surgery, Shanghai Tenth People’s Hospital, Tongji University, School of Medicine, Shanghai 200072, China
- Institute of Medical Intervention Engineering, Tongji University, Shanghai 200072, China
| | - Yinpeng Jin
- Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China
| | - Chuanwu Cao
- Department of Interventional & Vascular Surgery, Shanghai Tenth People’s Hospital, Tongji University, School of Medicine, Shanghai 200072, China
- Institute of Medical Intervention Engineering, Tongji University, Shanghai 200072, China
| | - Shilong Han
- Department of Interventional & Vascular Surgery, Shanghai Tenth People’s Hospital, Tongji University, School of Medicine, Shanghai 200072, China
- Institute of Medical Intervention Engineering, Tongji University, Shanghai 200072, China
| | - Xiaohu Yang
- Department of Interventional Radiology, Affiliated Hospital of Nantong University, Nantong 226001, China
| | - Suming Zhao
- Department of Interventional Radiology, Affiliated Hospital of Nantong University, Nantong 226001, China
| | - Maoquan Li
- Department of Interventional & Vascular Surgery, Shanghai Tenth People’s Hospital, Tongji University, School of Medicine, Shanghai 200072, China
- Institute of Medical Intervention Engineering, Tongji University, Shanghai 200072, China
| | - Hui Zhao
- Department of Interventional Radiology, Affiliated Hospital of Nantong University, Nantong 226001, China
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21
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Mao Y, Ma J, Xia Y, Xie X. The Overexpression of Epidermal Growth Factor (EGF) in HaCaT Cells Promotes the Proliferation, Migration, Invasion and Transdifferentiation to Epidermal Stem Cell Immunophenotyping of Adipose-Derived Stem Cells (ADSCs). Int J Stem Cells 2020; 13:93-103. [PMID: 32114740 PMCID: PMC7119215 DOI: 10.15283/ijsc18146] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/26/2018] [Revised: 05/28/2019] [Accepted: 05/28/2019] [Indexed: 11/09/2022] Open
Abstract
Background and Objectives The application of adipose derived stem cells (ADSCs) in skin repair has attracted much attention nowadays. Epidermal growth factor (EGF) participates in the progress of skin proliferation, differentiation and so forth. We aimed to explore the role of EGF in the proliferation, invasion, migration and transdifferentiation into epidermal cell phenotypes of ADSCs. Methods and Results ADSCs were extracted from adipose tissues from patient. Immunophenotyping was determined by flow cytometry. Overexpressed EGF or siEGF was transfected by lentiviruses. EGF was determined by enzyme linked immunosorbent assay (ELISA) or western blot. ADSCs and HaCaT cells were co-cultured by Transwell chambers. Conditioned medium (CM) was obtained from cultured HaCaT cells and used for the culturing of ADSCs. Cell viability was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Invasion rate was measured by Transwell invasion assay and migration rate by wound healing test. mRNA and protein levels were measured by qPCR and western blot respectively. The extracted cells from adipose tissues were identified as ADSCs by morphology and immunophenotyping. The expression of EGF was up or down regulated constantly in HaCaT cell line after transfection. EGF overexpression upregulated the proliferation, migration and invasion rates of ADSCs, and EGF expression regulated the expression of cytokeratin-19 (CK19) and integrin-β as well. Conclusions EGF could be served as a stimulus to promote the proliferation, migration, and invasion as well as the transdifferentiation into epidermal stem cell immunophenotyping of ADSCs. The results showed that EGF had a promising effect on the repair of skin wound.
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Affiliation(s)
- Yueping Mao
- Department of Dermatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China
| | - Jianchi Ma
- Department of Dermatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China
| | - Yue Xia
- Department of Dermatology, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
| | - Xiaoyuan Xie
- Department of Dermatology, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
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Zhong YC, Wang SC, Han YH, Wen Y. Recent Advance in Source, Property, Differentiation, and Applications of Infrapatellar Fat Pad Adipose-Derived Stem Cells. Stem Cells Int 2020; 2020:2560174. [PMID: 32215015 PMCID: PMC7081037 DOI: 10.1155/2020/2560174] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2019] [Revised: 02/12/2020] [Accepted: 02/20/2020] [Indexed: 12/18/2022] Open
Abstract
Infrapatellar fat pad (IPFP) can be easily obtained during knee surgery, which avoids the damage to patients for obtaining IPFP. Infrapatellar fat pad adipose-derived stem cells (IPFP-ASCs) are also called infrapatellar fat pad mesenchymal stem cells (IPFP-MSCs) because the morphology of IPFP-ASCs is similar to that of bone marrow mesenchymal stem cells (BM-MSCs). IPFP-ASCs are attracting more and more attention due to their characteristics suitable to regenerative medicine such as strong proliferation and differentiation, anti-inflammation, antiaging, secreting cytokines, multipotential capacity, and 3D culture. IPFP-ASCs can repair articular cartilage and relieve the pain caused by osteoarthritis, so most of IPFP-related review articles focus on osteoarthritis. This article reviews the anatomy and function of IPFP, as well as the discovery, amplification, multipotential capacity, and application of IPFP-ASCs in order to explain why IPFP-ASC is a superior stem cell source in regenerative medicine.
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Affiliation(s)
- Yu-chen Zhong
- Department of Histology and Embryology, College of Basic Medical Sciences, China Medical University, Shenyang 110122, China
- Class 4, Phase 102, China Medical University, Shenyang 110122, China
| | - Shi-chun Wang
- Department of Histology and Embryology, College of Basic Medical Sciences, China Medical University, Shenyang 110122, China
- Class 4, Phase 102, China Medical University, Shenyang 110122, China
| | - Yin-he Han
- Department of Histology and Embryology, College of Basic Medical Sciences, China Medical University, Shenyang 110122, China
| | - Yu Wen
- Department of Histology and Embryology, College of Basic Medical Sciences, China Medical University, Shenyang 110122, China
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Ni H, Zhao Y, Ji Y, Shen J, Xiang M, Xie Y. Adipose-derived stem cells contribute to cardiovascular remodeling. Aging (Albany NY) 2019; 11:11756-11769. [PMID: 31800397 PMCID: PMC6932876 DOI: 10.18632/aging.102491] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/14/2019] [Accepted: 11/17/2019] [Indexed: 02/06/2023]
Abstract
Obesity is an independent risk factor for cardiovascular disease. Adipose tissue was initially thought to be involved in metabolism through paracrine. Recent researches discovered mesenchymal stem cells inside adipose tissue which could differentiate into vascular lineages in vitro and in vivo, participating vascular remodeling. However, there were few researches focusing on distinct characteristics and functions of adipose-derived stem cells (ADSCs) from different regions. This is the first comprehensive review demonstrating the variances of ADSCs from the perspective of their origins.
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Affiliation(s)
- Hui Ni
- Department of Cardiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Yiming Zhao
- Department of Endocrinology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Yongli Ji
- Department of Cardiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Jian Shen
- Department of Cardiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Meixiang Xiang
- Department of Cardiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
| | - Yao Xie
- Department of Cardiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
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Pourang A, Rockwell H, Karimi K. New Frontiers in Skin Rejuvenation, Including Stem Cells and Autologous Therapies. Facial Plast Surg Clin North Am 2019; 28:101-117. [PMID: 31779934 DOI: 10.1016/j.fsc.2019.09.009] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023]
Abstract
One of the greatest challenges in the progression of aesthetic medicine lies in providing treatments with long-term results that are also minimally invasive and safe. Keeping up with this demand are developments in autologous therapies such as adipose-derived stem cells, stromal vascular fraction, microfat, nanofat, and platelet therapies, which are being shown to deliver satisfactory results. Innovations in more traditional cosmetic therapies, such as botulinum toxin, fillers, and thread lifts, are even more at the forefront of the advancement in aesthetics. Combining autologous therapies with traditional noninvasive methods can ultimately provide patients with more effective rejuvenation options.
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Affiliation(s)
- Aunna Pourang
- Department of Dermatology, University of California, Davis, 3301 C Street, Suite 1400, Sacramento, CA 95816, USA
| | - Helena Rockwell
- University of California, San Diego, School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093, USA
| | - Kian Karimi
- Rejuva Medical Aesthetics, 11645 Wilshire Boulevard #605, Los Angeles, CA 90025, USA.
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Isolation, Characterization, Differentiation and Immunomodulatory Capacity of Mesenchymal Stromal/Stem Cells from Human Perirenal Adipose Tissue. Cells 2019; 8:cells8111346. [PMID: 31671899 PMCID: PMC6928994 DOI: 10.3390/cells8111346] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2019] [Revised: 10/24/2019] [Accepted: 10/29/2019] [Indexed: 12/11/2022] Open
Abstract
Mesenchymal stromal/stem cells (MSCs) are immature multipotent cells, which represent a rare population in the perivascular niche within nearly all tissues. The most abundant source to isolate MSCs is adipose tissue. Currently, perirenal adipose tissue is rarely described as the source of MSCs. MSCs were isolated from perirenal adipose tissue (prASCs) from patients undergoing tumor nephrectomies, cultured and characterized by flow cytometry and their differentiation potential into adipocytes, chondrocytes, osteoblasts and epithelial cells. Furthermore, prASCs were stimulated with lipopolysaccharide (LPS), lipoteichoic acid (LTA) or a mixture of cytokines (cytomix). In addition, prASC susceptibility to human cytomegalovirus (HCMV) was investigated. The expression of inflammatory readouts was estimated by qPCR and immunoassay. HCMV infection was analyzed by qPCR and immunostaining. Characterization of cultured prASCs shows the cells meet the criteria of MSCs and prASCs can undergo trilineage differentiation. Cultured prASCs can be induced to differentiate into epithelial cells, shown by cytokeratin 18 expression. Stimulation of prASCs with LPS or cytomix suggests the cells are capable of initiating an inflammation-like response upon stimulation with LPS or cytokines, whereas, LTA did not induce a significant effect on the readouts (ICAM-1, IL-6, TNFα, MCP-1 mRNA and IL-6 protein). HCMV broadly infects prASCs, showing a viral load dependent cytopathological effect (CPE). Our current study summarizes the isolation and culture of prASCs, clearly characterizes the cells, and demonstrates their immunomodulatory potential and high permissiveness for HCMV.
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Hassanshahi M, Khabbazi S, Peymanfar Y, Hassanshahi A, Hosseini-Khah Z, Su YW, Xian CJ. Critical limb ischemia: Current and novel therapeutic strategies. J Cell Physiol 2019; 234:14445-14459. [PMID: 30637723 DOI: 10.1002/jcp.28141] [Citation(s) in RCA: 21] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/25/2018] [Accepted: 01/02/2019] [Indexed: 01/24/2023]
Abstract
Critical limb ischemia (CLI) is the advanced stage of peripheral artery disease spectrum and is defined by limb pain or impending limb loss because of compromised blood flow to the affected extremity. Current conventional therapies for CLI include amputation, bypass surgery, endovascular therapy, and pharmacological approaches. Although these conventional therapeutic strategies still remain as the mainstay of treatments for CLI, novel and promising therapeutic approaches such as proangiogenic gene/protein therapies and stem cell-based therapies have emerged to overcome, at least partially, the limitations and disadvantages of current conventional therapeutic approaches. Such novel CLI treatment options may become even more effective when other complementary approaches such as utilizing proper bioscaffolds are used to increase the survival and engraftment of delivered genes and stem cells. Therefore, herein, we address the benefits and disadvantages of current therapeutic strategies for CLI treatment and summarize the novel and promising therapeutic approaches for CLI treatment. Our analyses also suggest that these novel CLI therapeutic strategies show considerable advantages to be used when current conventional methods have failed for CLI treatment.
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Affiliation(s)
- Mohammadhossein Hassanshahi
- School of Pharmacy and Medical Sciences, University of South Australia Cancer Research Institute, University of South Australia, Adelaide, SA, Australia
| | - Samira Khabbazi
- School of Pharmacy and Medical Sciences, University of South Australia Cancer Research Institute, University of South Australia, Adelaide, SA, Australia
| | - Yaser Peymanfar
- School of Pharmacy and Medical Sciences, University of South Australia Cancer Research Institute, University of South Australia, Adelaide, SA, Australia
| | - Alireza Hassanshahi
- Department of Genetics, Faculty of Basic Sciences, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran
| | - Zahra Hosseini-Khah
- Department of Immunology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
| | - Yu-Wen Su
- School of Pharmacy and Medical Sciences, University of South Australia Cancer Research Institute, University of South Australia, Adelaide, SA, Australia
| | - Cory J Xian
- School of Pharmacy and Medical Sciences, University of South Australia Cancer Research Institute, University of South Australia, Adelaide, SA, Australia
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Pederzoli F, Joice G, Salonia A, Bivalacqua TJ, Sopko NA. Regenerative and engineered options for urethroplasty. Nat Rev Urol 2019; 16:453-464. [PMID: 31171866 DOI: 10.1038/s41585-019-0198-y] [Citation(s) in RCA: 32] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 05/04/2019] [Indexed: 02/07/2023]
Abstract
Surgical correction of urethral strictures by substitution urethroplasty - the use of grafts or flaps to correct the urethral narrowing - remains one of the most challenging procedures in urology and is frequently associated with complications, restenosis and poor quality of life for the affected individual. Tissue engineering using different cell types and tissue scaffolds offers a promising alternative for tissue repair and replacement. The past 30 years of tissue engineering has resulted in the development of several therapies that are now in use in the clinic, especially in treating cutaneous, bone and cartilage defects. Advances in tissue engineering for urethral replacement have resulted in several clinical applications that have shown promise but have not yet become the standard of care.
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Affiliation(s)
- Filippo Pederzoli
- Division of Experimental Oncology/Unit of Urology, URI, IRCCS Ospedale San Raffaele, Milan, Italy
- Department of Urology, James Buchanan Brady Urological Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA
- Università Vita-Salute San Raffaele, Milan, Italy
| | - Gregory Joice
- Department of Urology, James Buchanan Brady Urological Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA
| | - Andrea Salonia
- Division of Experimental Oncology/Unit of Urology, URI, IRCCS Ospedale San Raffaele, Milan, Italy
- Università Vita-Salute San Raffaele, Milan, Italy
| | - Trinity J Bivalacqua
- Department of Urology, James Buchanan Brady Urological Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA
| | - Nikolai A Sopko
- Department of Urology, James Buchanan Brady Urological Institute, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
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Secretome of Adipose Tissue-Derived Stem Cells (ASCs) as a Novel Trend in Chronic Non-Healing Wounds: An Overview of Experimental In Vitro and In Vivo Studies and Methodological Variables. Int J Mol Sci 2019; 20:ijms20153721. [PMID: 31366040 PMCID: PMC6696601 DOI: 10.3390/ijms20153721] [Citation(s) in RCA: 51] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2019] [Revised: 07/27/2019] [Accepted: 07/28/2019] [Indexed: 12/11/2022] Open
Abstract
Wound healing is a complex process with a linear development that involves many actors in a multistep timeline commonly divided into four stages: Hemostasis, inflammation, proliferation, and remodeling. Chronic non-healing wounds fail to progress beyond the inflammatory phase, thus precluding the next steps and, ultimately, wound repair. Many intrinsic or extrinsic factors may contribute to such an occurrence, including patient health conditions, age-related diseases, metabolic deficiencies, advanced age, mechanical pressure, and infections. Great interest is being focused on the adipose tissue-derived stem cell’s (ASC) paracrine activity for its potential therapeutic impact on chronic non-healing wounds. In this review, we summarize the results of in vitro and in vivo experimental studies on the pro-wound healing effects of ASC-secretome and/or extracellular vesicles (EVs). To define an overall picture of the available literature data, experimental conditions and applied methodologies are described as well as the in vitro and in vivo models chosen in the reported studies. Even if a comparative analysis of the results obtained by the different groups is challenging due to the large variability of experimental conditions, the available findings are undoubtedly encouraging and fully support the use of cell-free therapies for the treatment of chronic non-healing wounds.
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Giri TK, Alexander A, Agrawal M, Saraf S, Saraf S, Ajazuddin. Current Status of Stem Cell Therapies in Tissue Repair and Regeneration. Curr Stem Cell Res Ther 2019; 14:117-126. [PMID: 29732992 DOI: 10.2174/1574888x13666180502103831] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/26/2017] [Revised: 03/28/2018] [Accepted: 03/29/2018] [Indexed: 01/07/2023]
Abstract
Tissue engineering is a multi-disciplinary field such as material science, life science, and bioengineering that are necessary to make artificial tissue or rejuvenate damaged tissue. Numerous tissue repair techniques and substitute now exist even though it has several shortcomings; these shortcomings give a good reason for the continuous research for more acceptable tissue-engineered substitutes. The search for and use of a suitable stem cell in tissue engineering is a promising concept. Stem cells have a distinctive capability to differentiate and self-renew that make more suitable for cell-based therapies in tissue repair and regeneration. This review article focuses on stem cell for tissue engineering and their methods of manufacture with their application in nerve, bone, skin, cartilage, bladder, cardiac, liver tissue repair and regeneration.
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Affiliation(s)
- Tapan Kumar Giri
- NSHM College of Pharmaceutical Technology, NSHM Knowledge Campus, Kolkata Group of Institutions, 124 BL Saha Road, Kolkata-700053, West Bengal, India.,Rungta College of Pharmaceutical Sciences and Research, Bhilai, Chhattisgarh 490024, India
| | - Amit Alexander
- Rungta College of Pharmaceutical Sciences and Research, Bhilai, Chhattisgarh 490024, India
| | - Mukta Agrawal
- Rungta College of Pharmaceutical Sciences and Research, Bhilai, Chhattisgarh 490024, India
| | - Swarnalata Saraf
- Department of Pharmaceutics, University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh 492010, India
| | - Shailendra Saraf
- Department of Pharmaceutics, University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur, Chhattisgarh 492010, India.,Durg University, Govt. Vasudev Vaman Patankar Girls' P.G. College Campus, Raipur Naka, Durg, Chhattisgarh 491001, India
| | - Ajazuddin
- Rungta College of Pharmaceutical Sciences and Research, Bhilai, Chhattisgarh 490024, India
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30
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Zheng M, Kim DY, Sung JH. Ion channels and transporters in adipose-derived stem cells. JOURNAL OF PHARMACEUTICAL INVESTIGATION 2019. [DOI: 10.1007/s40005-018-00413-z] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/24/2023]
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Abstract
Aging is the result of two overlapping processes, "intrinsic" and "extrinsic." Intrinsic structural changes occur as a consequence of physiologic aging and are genetically determined; extrinsic relates to exposure to harmful events and habits, like smoking, bad diet, alcohol consumption, lack of sleep, stress, sun exposure, environmental pollution, etc. Aging may be decelerated by improving bad habits or treating signs of aging with various esthetic methods, food supplements, and antioxidants. It is believed that we cannot stop aging entirely due to the intrinsic part, which leads to irreversible cell damage, as well as tissue and organ damage due to their limited ability to regenerate. Stem cells and their ability to exhibit telomerase activity, to self-renew, and to differentiate into all three embryonic tissues challenges aging as a process, which is not inevitable and can even possibly be reversed. Stem cells can promote regeneration of aged tissues and organs by replacing apoptotic and necrotic cells with healthy ones. In addition, they can have antiinflammatory and antiapoptotic properties by paracrine-secreting growth factors and cytokines on the site of administration. Autologous adipose-derived stem cells are the most promising because they can be easily harvested in huge numbers with minimally invasive liposuction and, as such, represent a powerful tool in anti-aging and regenerative medicine. In this contribution, the author discusses their properties and application in clinical practice.
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32
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Natesan S, Wrice NL, Christy RJ. Peroxisome proliferator‐activated receptor‐α agonist and all‐
trans
retinoic acid induce epithelial differentiation of subcutaneous adipose‐derived stem cells from debrided burn skin. J Cell Biochem 2018; 120:9213-9229. [DOI: 10.1002/jcb.28197] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/10/2018] [Accepted: 11/12/2018] [Indexed: 11/06/2022]
Affiliation(s)
- Shanmugasundaram Natesan
- Combat Trauma and Burn Injury Research United States Army Institute of Surgical Research Fort Sam Houston Texas
| | - Nicole L. Wrice
- Combat Trauma and Burn Injury Research United States Army Institute of Surgical Research Fort Sam Houston Texas
| | - Robert J. Christy
- Combat Trauma and Burn Injury Research United States Army Institute of Surgical Research Fort Sam Houston Texas
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Wu Y, Kang YG, Cho H, Kim IG, Chung EJ, Shin JW. Combinational effects of mechanical forces and substrate surface characteristics on esophageal epithelial differentiation. J Biomed Mater Res A 2018; 107:552-560. [DOI: 10.1002/jbm.a.36571] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/06/2018] [Revised: 10/11/2018] [Accepted: 10/27/2018] [Indexed: 12/28/2022]
Affiliation(s)
- Yanru Wu
- Department of Health Science and Technology; Inje University; Gimhae Republic of Korea
| | - Yun Gyeong Kang
- Department of Biomedical Engineering; Inje University; Gimhae Republic of Korea
| | - Hana Cho
- Department of Otorhinolaryngology-Head and Neck Surgery; Seoul National University Hospital; Seoul Republic of Korea
| | - In Gul Kim
- Department of Otorhinolaryngology-Head and Neck Surgery; Seoul National University Hospital; Seoul Republic of Korea
| | - Eun-Jae Chung
- Department of Otorhinolaryngology-Head and Neck Surgery; Seoul National University Hospital; Seoul Republic of Korea
| | - Jung-Woog Shin
- Department of Health Science and Technology; Inje University; Gimhae Republic of Korea
- Department of Biomedical Engineering; Inje University; Gimhae Republic of Korea
- Cardiovascular and Metabolic Disease Center/Institute of Aged Life Redesign/UHARC, Inje University; Gimhae Republic of Korea
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Tong J, Mou S, Xiong L, Wang Z, Wang R, Weigand A, Yuan Q, Horch RE, Sun J, Yang J. Adipose-derived mesenchymal stem cells formed acinar-like structure when stimulated with breast epithelial cells in three-dimensional culture. PLoS One 2018; 13:e0204077. [PMID: 30335754 PMCID: PMC6193614 DOI: 10.1371/journal.pone.0204077] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2018] [Accepted: 08/31/2018] [Indexed: 02/08/2023] Open
Abstract
Lipotransfer has been applied in breast augmentation surgery for several years and the resident adipose-derived stem cells (ASCs) play an important role in enhancing fat graft survival. However, the interaction between ASCs and mammary epithelium is not fully understood. Many studies have shown that ASCs have a tumor-supportive effect in breast cancer. To the best of our knowledge, this is the first study on the effect of mammary epithelial cells on the human ASCs in 3D culture. ASCs were cultivated on matrigel in the conditioned medium (CM) prepared from a human breast epithelial cell line (HBL-100). The ASCs formed KRT18-positive acini-like structures after stimulation with breast epithelial cells. The expression of epithelial genes (CDH1 and KRT18) was up-regulated while the expression of mesenchymal specific genes (CDH2 and VIM) was down-regulated as determined by qRT-PCR. The stemness marker (CD29) and angiogenic factors (CD31 and VEGF) were also down-regulated as examined by immunofluorescence. In addition, the CM obtained from HBL-100 enhanced the migration and inhibited the adipogenic differentiation of ASCs. These results demonstrate that ASCs have the ability to transform into epithelial-like cells when cultured with mammary epithelial cells. Given these observations, we infer that ASCs have a positive effect on lipotransfer, not only due to their ability to secrete growth factors, but also due to their direct participation in the formation of new breast tissue.
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Affiliation(s)
- Jing Tong
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Shan Mou
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Lingyun Xiong
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Zhenxing Wang
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Rongrong Wang
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Annika Weigand
- Department of Plastic and Hand Surgery and Laboratory for Tissue Engineering and Regenerative Medicine, University Hospital Erlangen, Friedrich Alexander University, Erlangen‐Nuernberg, FAU, Germany
| | - Quan Yuan
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Raymund E. Horch
- Department of Plastic and Hand Surgery and Laboratory for Tissue Engineering and Regenerative Medicine, University Hospital Erlangen, Friedrich Alexander University, Erlangen‐Nuernberg, FAU, Germany
| | - Jiaming Sun
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
- * E-mail: (JY); (JS)
| | - Jie Yang
- Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
- * E-mail: (JY); (JS)
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Doornaert M, Colle J, De Maere E, Declercq H, Blondeel P. Autologous fat grafting: Latest insights. Ann Med Surg (Lond) 2018; 37:47-53. [PMID: 30622707 PMCID: PMC6318549 DOI: 10.1016/j.amsu.2018.10.016] [Citation(s) in RCA: 53] [Impact Index Per Article: 7.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/05/2018] [Revised: 10/06/2018] [Accepted: 10/11/2018] [Indexed: 12/13/2022] Open
Abstract
A recent rise in the use of autologous fat transfer for soft tissue augmentation has paralleled the increasing popularity of liposuction body contouring. This creates a readily available and inexpensive product for lipografting, which is the application of lipoaspirated material. Consistent scientific proof about the long-term viability of the transferred fat is not available. Clinically, there is a reabsorption rate which has been reported to range from 20 to 90%. Results can be unpredictable with overcorrection and regular need for additional interventions. In this review, adipogenesis physiology and the adipogenic cascade from adipose-derived stem cells to adult adipocytes is extensively described to determine various procedures involved in the fat grafting technique. Variables in structure and physiology, adipose tissue harvesting- and processing techniques, and the preservation of fat grafts are taken into account to collect reproducible scientific data to establish standard in vitro and in vivo models for experimental fat grafting. Adequate histological staining for fat tissue, immunohistochemistry and viability assays should be universally used in experiments to be able to produce comparative results. By analysis of the applied methods and comparison to similar experiments, a conclusion concerning the ideal technique to improve clinical outcome is proposed.
Adipogenic physiology is described to determine various procedures involved in the fat grafting technique. Clinical studies on fat grafting have confirmed an unpredictable result. After analysis of the literature and despite attempts to eliminate confounding factors, on every step of the fat transfer technique a number of studies with conflicting results exist. Adequate histological staining for fat tissue, immunohistochemistry and viability assays should be universally used in experiments to be able to produce comparative results.
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36
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Thelen K, Watts SW, Contreras GA. Adipogenic potential of perivascular adipose tissue preadipocytes is improved by coculture with primary adipocytes. Cytotechnology 2018; 70:1435-1445. [PMID: 30051281 PMCID: PMC6214855 DOI: 10.1007/s10616-018-0238-0] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/23/2018] [Accepted: 07/19/2018] [Indexed: 12/21/2022] Open
Abstract
Perivascular adipose tissue (PVAT) has the capacity to secrete vasoactive mediators with the potential to regulate vascular function. Given its location adjacent to the vasculature, PVAT dysfunction may be part of the pathophysiology of cardiovascular diseases. To study the mechanisms of PVAT dysfunction, several adipogenic models have been proposed. However, these approaches do not adequately reflect PVAT adipocyte phenotypes variability that depends on their anatomical location. Despite PVAT importance in modulating vascular function, to date, there is not a depot-specific adipogenic model for PVAT adipocytes. We present a model that uses coculturing of PVAT stromal vascular fraction derived preadipocytes with primary adipocytes isolated from the same PVAT. Preadipocytes were isolated from thoracic aorta PVAT and mesenteric resistance artery PVAT (mPVAT). Upon confluency, cells were induced to differentiate for 7 and 14 days using a standard protocol (SP) or standard protocol cocultured with primary adipocytes isolated from the same adipose depots (SPA) for 96, 120, and 144 h. SPA reduced the time for differentiation of stromal vascular fraction derived preadipocytes and increased their capacity to store lipids compared with SP as indicated by lipid accumulation, lipolytic responses, gene marker profile expression, and adiponectin secretion. The coculture system improved adipogenesis efficiency by enhancing lipid accumulation and reducing the time of induction, therefore, is a more efficient method compared to SP alone.
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Affiliation(s)
- Kyan Thelen
- Department of Large Animal Clinical Sciences, Michigan State University, 736 Wilson Road, Room D202, East Lansing, MI, 48824, USA
| | - Stephanie W Watts
- Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI, USA
| | - G Andres Contreras
- Department of Large Animal Clinical Sciences, Michigan State University, 736 Wilson Road, Room D202, East Lansing, MI, 48824, USA.
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37
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La Padula S, Hersant B, Meningaud J, D’Andrea F. Use of autologous fat graft and fractiononal co2 laser to optimize the aesthetic and functional results in patients with severe burn outcomes of the face. JOURNAL OF STOMATOLOGY, ORAL AND MAXILLOFACIAL SURGERY 2018; 119:279-283. [DOI: 10.1016/j.jormas.2018.04.005] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/01/2018] [Accepted: 04/12/2018] [Indexed: 11/30/2022]
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Hintze JM, Tchoukalova YD, Sista R, Shah MK, Zhang N, Lott DG. Development of xeno-free epithelial differentiation media for adherent, non-expanded adipose stromal vascular cell cultures. Biochem Biophys Res Commun 2018; 503:3128-3133. [PMID: 30166060 DOI: 10.1016/j.bbrc.2018.08.104] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2018] [Accepted: 08/15/2018] [Indexed: 12/12/2022]
Abstract
INTRODUCTION Reconstruction of respiratory epithelium is critical for the fabrication of bioengineered airway implants. Epithelial differentiation is typically achieved using bovine pituitary extract (BPE). Due to the xenogenic nature and undefined composition of BPE, an alternative for human clinical applications, devoid of BPE, must be developed. The goal of this study was to develop two different BPE-free media, with and without select pituitary hormone (PH), which could initiate epithelial differentiation for use in human implantation. METHODS The ability of the two BPE-free media to initiate epithelial differentiation of adherent, non-expanded stromal-vascular cells grown on porcine small intestinal submucosa was compared to traditional BPE-containing media (M1). Nanostring® was used to measure differences in gene expression of stemness (MSC), basal cell (basal), and ciliated markers (muco-cil), and staining was performed support the gene data. RESULTS Compared to baseline, both BPE-free media upregulated epithelial and stemness genes, however this was to a lower degree than BPE-containing media. In general, the expression of basal cell markers (COL17A1, DSG3, ITGA6, KRT6A, LOXL2) and secreted mucous proteins (PLUNC, MUC5B, SCGB2A1) was upregulated. The gene expression of ciliated markers C9orf24, TUBA3 and DNCL2B but not of the key transcription factor for cilagenesis FOXJ1 were upregulated, indicating that mucus-secreting cell differentiation occurs more rapidly than ciliogenesis. The ability of the adherent stromal vascular cells to upregulate gene expression of both epithelial and stemness markers suggests maintenance of the self-renewal capacity of undifferentiated and/or basal cell-like cells contributing to proliferation and ensuring a persisting source of cells for regenerative medicine applications. CONCLUSION This study provides the initial step to defining a BPE-free epithelial differentiation medium for clinical translation. Thus, either of the proposed BPE-free medium are viable alternatives to BPE-containing medium for partial epithelial differentiation for human translational applications.
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Affiliation(s)
- Justin M Hintze
- Head and Neck Regeneration Program, Center for Regenerative Medicine, Mayo Clinic, USA
| | - Yourka D Tchoukalova
- Head and Neck Regeneration Program, Center for Regenerative Medicine, Mayo Clinic, USA
| | - Ramachandra Sista
- Department of Pulmonary and Critical Care Medicine, Mayo Clinic, Phoenix, AZ, USA
| | - Manisha K Shah
- Head and Neck Regeneration Program, Center for Regenerative Medicine, Mayo Clinic, USA
| | - Nan Zhang
- Department of Biostatistics, Mayo Clinic, Scottsdale, AZ, USA
| | - David G Lott
- Head and Neck Regeneration Program, Center for Regenerative Medicine, Mayo Clinic, USA; Department of Otolaryngology - Head and Neck Surgery, Mayo Clinic, Phoenix, AZ, USA.
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Dubey NK, Mishra VK, Dubey R, Deng YH, Tsai FC, Deng WP. Revisiting the Advances in Isolation, Characterization and Secretome of Adipose-Derived Stromal/Stem Cells. Int J Mol Sci 2018; 19:ijms19082200. [PMID: 30060511 PMCID: PMC6121360 DOI: 10.3390/ijms19082200] [Citation(s) in RCA: 80] [Impact Index Per Article: 11.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2018] [Revised: 07/08/2018] [Accepted: 07/24/2018] [Indexed: 12/13/2022] Open
Abstract
Adipose-derived stromal/stem cells (ASCs) seems to be a promising regenerative therapeutic agent due to the minimally invasive approach of their harvest and multi-lineage differentiation potential. The harvested adipose tissues are further digested to extract stromal vascular fraction (SVF), which is cultured, and the anchorage-dependent cells are isolated in order to characterize their stemness, surface markers, and multi-differentiation potential. The differentiation potential of ASCs is directed through manipulating culture medium composition with an introduction of growth factors to obtain the desired cell type. ASCs have been widely studied for its regenerative therapeutic solution to neurologic, skin, wound, muscle, bone, and other disorders. These therapeutic outcomes of ASCs are achieved possibly via autocrine and paracrine effects of their secretome comprising of cytokines, extracellular proteins and RNAs. Therefore, secretome-derivatives might offer huge advantages over cells through their synthesis and storage for long-term use. When considering the therapeutic significance and future prospects of ASCs, this review summarizes the recent developments made in harvesting, isolation, and characterization. Furthermore, this article also provides a deeper insight into secretome of ASCs mediating regenerative efficacy.
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Affiliation(s)
- Navneet Kumar Dubey
- Ceramics and Biomaterials Research Group, Advanced Institute of Materials Science, Ton Duc Thang University, Ho Chi Minh City 700000, Vietnam.
- Faculty of Applied Sciences, Ton Duc Thang University, Ho Chi Minh City 700000, Vietnam.
| | - Viraj Krishna Mishra
- Applied Biotech Engineering Centre (ABEC), Department of Biotechnology, Ambala College of Engineering and Applied Research, Ambala 133101, India.
| | - Rajni Dubey
- Graduate Institute Food Science and Technology, National Taiwan University, Taipei 10617, Taiwan.
| | - Yue-Hua Deng
- Stem Cell Research Center, Taipei Medical University, Taipei 11031, Taiwan.
- Department of Life Science, Fu Jen Catholic University, New Taipei City 24205, Taiwan.
| | - Feng-Chou Tsai
- School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei 11031, Taiwan.
| | - Win-Ping Deng
- Stem Cell Research Center, Taipei Medical University, Taipei 11031, Taiwan.
- School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei 11031, Taiwan.
- Department of Basic medicine, Fu-Jen Catholic University, New Taipei City 24205, Taiwan.
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Badimon L, Cubedo J. Adipose tissue depots and inflammation: effects on plasticity and resident mesenchymal stem cell function. Cardiovasc Res 2018; 113:1064-1073. [PMID: 28498891 DOI: 10.1093/cvr/cvx096] [Citation(s) in RCA: 79] [Impact Index Per Article: 11.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/30/2017] [Accepted: 05/10/2017] [Indexed: 02/07/2023] Open
Abstract
Adipose tissue (AT) is a highly heterogeneous organ. Beside the heterogeneity associated to different tissue types (white, brown, and 'brite') and its location-related heterogeneity (subcutaneous, visceral, epicardial, and perivascular, etc.), AT composition, structure, and functionality are highly dependent on individual-associated factors. As such, the pro-inflammatory state associated to the presence of obesity and other cardiovascular risk factors (CVRFs) directly affects AT metabolism. Furthermore, the adipose-derived stem cells (ASCs) that reside in the stromal vascular fraction of AT, besides being responsible for most of the plasticity attributed to AT, is an additional source of heterogeneity. Thus, ASCs directly contribute to AT homeostasis, cell renewal, and spontaneous repair. These ASCs share many properties with the bone-marrow mesenchymal stem cells (i.e. potential to differentiate towards multiple tissue lineages, and angiogenic, antiapoptotic, and immunomodulatory properties). Moreover, ASCs show clear advantages in terms of accessibility and quantity of available sample, their easy in vitro expansion, and the possibility of having an autologous source. All these properties point out towards a potential use of ASCs in regenerative medicine. However, the presence of obesity and other CVRFs induces a pro-inflammatory state that directly impacts ASCs proliferation and differentiation capacities affecting their regenerative abilities. The focus of this review is to summarize how inflammation affects the different AT depots and the mechanisms by which these changes further enhance the obesity-associated metabolic disturbances. Furthermore, we highlight the impact of obesity-induced inflammation on ASCs properties and how those effects impair their plasticity.
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Affiliation(s)
- Lina Badimon
- Cardiovascular Science Institute - ICCC, IIB-Sant Pau, CiberCV, Hospital de Sant Pau, c/Sant Antoni M Claret 167, Barcelona 08025, Spain.,Cardiovascular Research Chair UAB, Barcelona, Spain
| | - Judit Cubedo
- Cardiovascular Science Institute - ICCC, IIB-Sant Pau, CiberCV, Hospital de Sant Pau, c/Sant Antoni MaClaret 167, Barcelona 08025, Spain
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Composite Scaffolds Based on Intestinal Extracellular Matrices and Oxidized Polyvinyl Alcohol: A Preliminary Study for a New Regenerative Approach in Short Bowel Syndrome. BIOMED RESEARCH INTERNATIONAL 2018; 2018:7824757. [PMID: 29992163 PMCID: PMC5994320 DOI: 10.1155/2018/7824757] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/30/2017] [Accepted: 04/17/2018] [Indexed: 12/11/2022]
Abstract
Pediatric Short Bowel Syndrome is a rare malabsorption disease occurring because of massive surgical resections of the small intestine. To date, the issues related to current strategies including intestinal transplantation prompted the attention towards tissue engineering (TE). This work aimed to develop and compare two composite scaffolds for intestinal TE consisting of a novel hydrogel, that is, oxidized polyvinyl alcohol (OxPVA), cross-linked with decellularized intestinal wall as a whole (wW/OxPVA) or homogenized (hW/OxPVA). A characterization of the supports was performed by histology and Scanning Electron Microscopy and their interaction with adipose mesenchymal stem cells occurred by MTT assay. Finally, the scaffolds were implanted in the omentum of Sprague Dawley rats for 4 weeks prior to being processed by histology and immunohistochemistry (CD3; F4/80; Ki-67; desmin; α-SMA; MNF116). In vitro studies proved the effectiveness of the decellularization, highlighting the features of the matrices; moreover, both supports promoted cell adhesion/proliferation even if the wW/OxPVA ones were more effective (p < 0.01). Analysis of explants showed a continuous and relatively organized tissue wall around the supports with a connective appearance, such as myofibroblastic features, smooth muscle, and epithelial cells. Both scaffolds, albeit with some difference, were promising; nevertheless, further analysis will be necessary.
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Li Y, Shan Z, Yang B, Yang D, Men C, Cui Y, Wu J. Cathelicidin LL37 Promotes Epithelial and Smooth-Muscle-Like Differentiation of Adipose-Derived Stem Cells through the Wnt/β-Catenin and NF-κB Pathways. BIOCHEMISTRY (MOSCOW) 2018; 82:1336-1345. [PMID: 29223160 DOI: 10.1134/s0006297917110116] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2023]
Abstract
Ureter reconstruction is a difficult procedure in urology. Adipose-derived stem cells (ADSCs), along with multipotency and self-renewal capacity, are a preferred choice for tissue engineering-based ureteral reconstruction. We explored the synergic role of cathelicidin LL37 (LL37) in epithelial and smooth-muscle-like differentiation. ADSCs were separated from adipose tissues of mouse and characterized by flow cytometry. The ADSCs were then stably transfected with pGC-FU-GFP (pGC) or pGC containing full-length LL37 (pGC-LL37), respectively. Cell viability and apoptosis were respectively estimated in the stably transfected cells and non-transfected cells. Then, qRT-PCR and Western blot analysis were used for determinations of epithelial marker expressions after induction by all-trans retinoic acid as well as smooth-muscle-like marker expressions after induction by transforming growth factor-β1. Then, possibly involved signaling pathways and extracellular expression of LL37 were detected. Cell viability and apoptosis were not changed after LL37 overexpression. Expression levels of epithelial and smooth-muscle-like markers were significantly upregulated by LL37 overexpression. Moreover, expressions of key kinases involved in the Wnt/β-catenin pathway as well as epithelial marker were upregulated by the LL37 overexpression, while it was reversed by Wnt/β-catenin inhibitor. Likewise, expressions of key kinases involved in the nuclear factor κB (NF-κB) pathway as well as smooth-muscle-like markers were upregulated by LL37 overexpression, which was reversed by NF-κB inhibitor. LL37 was found in the culture medium. LL37, which could be released into the medium, had no impact on cell proliferation and apoptosis of ADSCs. However, LL37 promoted epithelial and smooth-muscle-like differentiation through activating the Wnt/β-catenin and NF-κB pathways, respectively.
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Affiliation(s)
- Yongwei Li
- Department of Urology Surgery, Qingdao University, Affiliated Yantai Yuhuangding Hospital, Yantai, 264000, China.
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Choi B, Kim D, Han I, Lee SH. Microenvironmental Regulation of Stem Cell Behavior Through Biochemical and Biophysical Stimulation. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2018; 1064:147-160. [PMID: 30471031 DOI: 10.1007/978-981-13-0445-3_9] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/22/2022]
Abstract
Stem cells proliferate by undergoing self-renewal and differentiate into multiple cell lineages in response to biochemical and biophysical stimuli. Various biochemical cues such as growth factors, nucleic acids, chemical reagents, and small molecules have been used to induce stem cell differentiation or reprogramming or to maintain their pluripotency. Moreover, biophysical cues such as matrix stiffness, substrate topography, and external stress and strain play a major role in modulating stem cell behavior. In this chapter, we have summarized microenvironmental regulation of stem cell behavior through biochemical and biophysical stimulation.
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Affiliation(s)
- Bogyu Choi
- Department of Biomedical Science, CHA University, Seongnam-si, South Korea
| | - Deogil Kim
- Department of Biomedical Science, CHA University, Seongnam-si, South Korea
| | - Inbo Han
- Department of Neurosurgery, CHA University, CHA Bundang Medical Center, Seongnam-si, South Korea
| | - Soo-Hong Lee
- Department of Medical Biotechnology, Dongguk University, Goyang-si, Gyeonggi-do, South Korea.
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RUNX2 promotes epithelial differentiation of ADSCs and burn wound healing via targeting E-cadherin. Oncotarget 2017; 9:2646-2659. [PMID: 29416798 PMCID: PMC5788666 DOI: 10.18632/oncotarget.23522] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/06/2017] [Accepted: 09/21/2017] [Indexed: 12/11/2022] Open
Abstract
Epithelial differentiation of adipose-derived stem cells (ADSCs) is mediated by sophisticated interactions of various molecular functions and biological processes, including transcriptional regulation. Runt-related transcription factor 2 (RUNX2) increases osteoblast and adipocyte differentiation of ADSCs. However, the role of RUNX2 in epithelial differentiation of ADSCs is unknown. We first showed that ADSCs possess the potential to differentiate into epithelial lineage. Then, we employed the effect of RUNX2 on epithelial differentiation of ADSCs. Our data showed that RUNX2 promoted epithelial differentiation of ADSCs. Overexpression or knockdown of RUNX2 resulted in increase or decrease of E-cadherin expression, respectively. Abatement of E-cadherin in ADSCs attenuated RUNX2-activated epithelial conversion of ADSCs and epithelial markers cytokeratin 18 (CK18) and zonula occludens protein-1 (ZO-1). We also evaluated the effect of RUNX2 on burn wound healing in vivo. The wound re-epithelialization were accelerated by RUNX2. The wound closure indexs, demis regeneration and revascularization were all improved. Furthermore, RUNX2 binding directly to the E-cadherin promoter region was characterized in ADSCs by chromatin immunoprecipitation (ChIP) and luciferase promoter reporter assays. Taken together, the study demonstrates the role of RUNX2 in epithelial differentiation of ADSCs and suggests that RUNX2 promotes E-cadherin expression, at least in part, through its direct binding to the promoter.
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Sola A, Saenz Del Burgo L, Ciriza J, Hernandez RM, Orive G, Martin Cordero J, Calle P, Pedraz JL, Hotter G. Microencapsulated macrophages releases conditioned medium able to prevent epithelial to mesenchymal transition. Drug Deliv 2017; 25:91-101. [PMID: 29250977 PMCID: PMC6058712 DOI: 10.1080/10717544.2017.1413449] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/02/2022] Open
Abstract
Epithelial to mesenchymal transition (EMT) has emerged as a key process in the development of renal fibrosis. In fact, EMT-derived fibroblasts contribute to the progression of chronic renal disease. In addition, anti-inflammatory M2 macrophages have exhibited a great influence on renal fibrosis. However, because of the high impact that the inputs of different environmental cytokines have on their phenotype, macrophages can easily lose this property. We aim to known if microencapsulated macrophages on M2-inducing alginate matrices could preserve macrophage phenotype and thus release factors able to act on epithelial cells to prevent the epithelial differentiation towards mesenchymal cells. We reproduced an in vitro model of EMT by treating adipose-derived stem cells with all-trans retinoic acid (ATRA) and induced their transformation toward epithelia. Dedifferentiation of epithelial cells into a mesenchymal phenotype occurred when ATRA was retired, thus simulating EMT. Results indicate that induction of M2 phenotype by IL-10 addition in the alginate matrix produces anti-inflammatory cytokines and increases the metabolic activity and the viability of the encapsulated macrophages. The released conditioned medium modulates EMT and maintains healthy epithelial phenotype. This could be used for in vivo cell transplantation, or alternatively as an external releaser able to prevent epithelial to mesenchymal transformation for future anti-fibrotic therapies.
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Affiliation(s)
- Anna Sola
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain
| | - Laura Saenz Del Burgo
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain.,b NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy , University of the Basque Country (UPV/EHU) , Vitoria-Gasteiz , Spain
| | - Jesús Ciriza
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain.,b NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy , University of the Basque Country (UPV/EHU) , Vitoria-Gasteiz , Spain
| | - Rosa Maria Hernandez
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain.,b NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy , University of the Basque Country (UPV/EHU) , Vitoria-Gasteiz , Spain
| | - Gorka Orive
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain.,b NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy , University of the Basque Country (UPV/EHU) , Vitoria-Gasteiz , Spain
| | - Jorge Martin Cordero
- c Department of Experimental Pathology , Instituto de Investigaciones Biomédicas de Barcelona, Spanish Research Council (IIBB-CSIC, IDIBAPS) , Barcelona , Spain
| | - Priscila Calle
- c Department of Experimental Pathology , Instituto de Investigaciones Biomédicas de Barcelona, Spanish Research Council (IIBB-CSIC, IDIBAPS) , Barcelona , Spain
| | - Jose Luis Pedraz
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain.,b NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy , University of the Basque Country (UPV/EHU) , Vitoria-Gasteiz , Spain
| | - Georgina Hotter
- a Biomedical Research Networking Center in Bioengineering , Biomaterials and Nanomedicine (CIBER-BBN) , Barcelona , Spain.,c Department of Experimental Pathology , Instituto de Investigaciones Biomédicas de Barcelona, Spanish Research Council (IIBB-CSIC, IDIBAPS) , Barcelona , Spain
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Abstract
Adipose-derived stem/stromal cells (ASCs), together with adipocytes, vascular endothelial cells, and vascular smooth muscle cells, are contained in fat tissue. ASCs, like the human bone marrow stromal/stem cells (BMSCs), can differentiate into several lineages (adipose cells, fibroblast, chondrocytes, osteoblasts, neuronal cells, endothelial cells, myocytes, and cardiomyocytes). They have also been shown to be immunoprivileged, and genetically stable in long-term cultures. Nevertheless, unlike the BMSCs, ASCs can be easily harvested in large amounts with minimal invasive procedures. The combination of these properties suggests that these cells may be a useful tool in tissue engineering and regenerative medicine.
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Affiliation(s)
- Simone Ciuffi
- Department of Surgery and Translational Medicine, University of Florence, Florence, Italy
| | - Roberto Zonefrati
- Department of Surgery and Translational Medicine, University of Florence, Florence, Italy
| | - Maria Luisa Brandi
- Department of Surgery and Translational Medicine, University of Florence, Florence, Italy
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Kim YJ, Park SG, Shin B, Kim J, Kim SW, Choo OS, Yin XY, Min BH, Choung YH. Osteogenesis for postoperative temporal bone defects using human ear adipose-derived stromal cells and tissue engineering: An animal model study. J Biomed Mater Res A 2017; 105:3493-3501. [PMID: 28875515 DOI: 10.1002/jbm.a.36194] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/23/2017] [Revised: 07/12/2017] [Accepted: 08/16/2017] [Indexed: 02/06/2023]
Abstract
Mastoidectomy, the removal of infected mastoid bones, is a common surgical procedure for the treatment of chronic otitis media. Persistent and recurrent otorrhea and accumulation of keratin debris following open cavity mastoidectomy are still bothersome issues for both patients and otologists. In this study, we used human ear adipose-derived stromal cells (hEASCs) in combination with polycaprolactone (PCL) scaffolds and osteogenic differentiation medium (ODM) to regenerate temporal bone defects. The hEASCs showed stem cell phenotypes, and these characteristics were maintained up to passage 5. Mastoid bulla and cranial bone defects were induced in Sprague-Dawley rats using AgNO3 and burr hole drilling, respectively, and the rats were then divided into five groups: (1) control, (2) hEASCs, (3) hEASCs + ODM, (4) hEASCs + PCL scaffolds, and (5) hEASCs + PCL scaffolds + ODM. Osteogenesis was evaluated by micro-computed tomography and histology. Compared with the control group, the groups transplanted with hEASCs and PCL scaffolds had significantly higher bone formation along the periphery of the mastoid bulla area. Moreover, ODM synergistically enhanced bone formation in mastoid bulla defects. Our results suggest that combining hEASCs with PCL scaffolds represents a promising method for anatomical and functional reconstruction of postoperative temporal bone defects following mastoidectomy. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3493-3501, 2017.
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Affiliation(s)
- Yeon Ju Kim
- Department of Otolaryngology, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
| | - Seung Gu Park
- Department of Otolaryngology, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
| | - Beomyong Shin
- Department of Otolaryngology, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea.,Department of Biomedical Sciences, BK21 Plus Research Center for Biomedical Sciences, Ajou University Graduate School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
| | - Jangho Kim
- Department of Rural and Biosystems Engineering, Chonnam National University, Gwangju, 61186, Republic of Korea
| | - Seung Won Kim
- Department of Burns and Plastic Surgery, Affiliated Hospital of Yanbian University, 1327 Juzi Street, Yanji, Jilin, 133000, China
| | - Oak-Sung Choo
- Department of Otolaryngology, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea.,Department of Medical Sciences, Ajou University Graduate School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
| | - Xiang Yun Yin
- Department of Orthopedic Surgery, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
| | - Byoung Hyun Min
- Department of Orthopedic Surgery, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
| | - Yun-Hoon Choung
- Department of Otolaryngology, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea.,Department of Biomedical Sciences, BK21 Plus Research Center for Biomedical Sciences, Ajou University Graduate School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea.,Department of Medical Sciences, Ajou University Graduate School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea
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Application of adipose-derived stromal cells in fat grafting: Basic science and literature review. Exp Ther Med 2017; 14:2415-2423. [PMID: 28962175 PMCID: PMC5609216 DOI: 10.3892/etm.2017.4811] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2016] [Accepted: 01/06/2017] [Indexed: 02/07/2023] Open
Abstract
Autologous fat is considered the ideal material for soft-tissue augmentation in plastic and reconstructive surgery. The primary drawback of autologous fat grafting is the high resorption rate. The isolation of mesenchymal stem cells from adipose tissue inevitably led to research focusing on the study of combined transplantation of autologous fat and adipose derived stem cells (ADSCs) and introduced the theory of ‘cell-assisted lipotransfer’. Transplantation of ADSCs is a promising strategy, due to the high proliferative capacity of stem cells, their potential to induce paracrine signalling and ability to differentiate into adipocytes and vascular cells. The current study examined the literature for clinical and experimental studies on cell-assisted lipotransfer to assess the efficacy of this novel technique when compared with traditional fat grafting. A total of 30 studies were included in the present review. The current study demonstrates that cell-assisted lipotransfer has improved efficacy compared with conventional fat grafting. Despite relatively positive outcomes, further investigation is required to establish a consensus in cell-assisted lipotransfer.
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Bertozzi N, Simonacci F, Grieco MP, Grignaffini E, Raposio E. The biological and clinical basis for the use of adipose-derived stem cells in the field of wound healing. Ann Med Surg (Lond) 2017; 20:41-48. [PMID: 28702186 PMCID: PMC5491486 DOI: 10.1016/j.amsu.2017.06.058] [Citation(s) in RCA: 67] [Impact Index Per Article: 8.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/12/2017] [Revised: 06/21/2017] [Accepted: 06/21/2017] [Indexed: 02/08/2023] Open
Abstract
Worldwide, hard-to-heal lower limb wounds are estimated to affect 1.5–3% of the adult population with a treatment-related annual cost of $10 billion. Thus, chronic skin ulcers of the lower limb are a matter of economic and public concern. Over the years, multiple medical and surgical approaches have been proposed but they are still inadequate, and no effective therapy yet exists. Regenerative medicine and stem cell-based therapies hold great promise for wound healing. Recently, many plastic surgeons have studied the potential clinical application of adipose-derived stem cells (ASCs), which are a readily available adult stem cell population that can undergo multilineage differentiation and secrete growth factors that can enhance wound-healing processes by promoting angiogenesis, and hence increase local blood supply. ASCs have been widely studied in vitro and in vivo in animal models. However, there are few randomized clinical trials on humans, and these are still ongoing or recruiting patients. Moreover, there is no consensus on a common isolation protocol that is clinically feasible and which would ensure reproducible results. The authors aim to provide readers with an overview of the biological properties of ASCs as well as their clinical application, to help better understanding of present and future strategies for the treatment of hard-to-heal wounds by means of stem cell-based therapies.
Worldwide, hard-to-heal wounds are a matter of economic and public concern. The emerging fields of regenerative medicine and stem cell-based therapies hold great promise for wound healing. ASCs can potentially give the support necessary for recovery of hard-to-heal wounds. ASCs can be easily harvested from adipose tissue by means of standard wet liposuction technique. ASCs have been widely studied in vitro and in vivo to demonstrate their potential and safety.
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Affiliation(s)
- Nicolò Bertozzi
- Department of Medicine and Surgery, Plastic Surgery Division, University of Parma, Via Gramsci, 14, 43126, Parma, Italy.,Cutaneous, Mininvasive, Regenerative and Plastic Surgery Unit, Parma University Hospital, Via Gramsci, 14, 43126, Parma, Italy
| | - Francesco Simonacci
- Department of Medicine and Surgery, Plastic Surgery Division, University of Parma, Via Gramsci, 14, 43126, Parma, Italy.,Cutaneous, Mininvasive, Regenerative and Plastic Surgery Unit, Parma University Hospital, Via Gramsci, 14, 43126, Parma, Italy
| | - Michele Pio Grieco
- Department of Medicine and Surgery, Plastic Surgery Division, University of Parma, Via Gramsci, 14, 43126, Parma, Italy.,Cutaneous, Mininvasive, Regenerative and Plastic Surgery Unit, Parma University Hospital, Via Gramsci, 14, 43126, Parma, Italy
| | - Eugenio Grignaffini
- Department of Medicine and Surgery, Plastic Surgery Division, University of Parma, Via Gramsci, 14, 43126, Parma, Italy.,Cutaneous, Mininvasive, Regenerative and Plastic Surgery Unit, Parma University Hospital, Via Gramsci, 14, 43126, Parma, Italy
| | - Edoardo Raposio
- Department of Medicine and Surgery, Plastic Surgery Division, University of Parma, Via Gramsci, 14, 43126, Parma, Italy.,Cutaneous, Mininvasive, Regenerative and Plastic Surgery Unit, Parma University Hospital, Via Gramsci, 14, 43126, Parma, Italy
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50
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Zou Q, Fu Q. Tissue engineering for urinary tract reconstruction and repair: Progress and prospect in China. Asian J Urol 2017; 5:57-68. [PMID: 29736367 PMCID: PMC5934513 DOI: 10.1016/j.ajur.2017.06.010] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2016] [Revised: 03/10/2017] [Accepted: 04/25/2017] [Indexed: 12/11/2022] Open
Abstract
Several urinary tract pathologic conditions, such as strictures, cancer, and obliterations, require reconstructive plastic surgery. Reconstruction of the urinary tract is an intractable task for urologists due to insufficient autologous tissue. Limitations of autologous tissue application prompted urologists to investigate ideal substitutes. Tissue engineering is a new direction in these cases. Advances in tissue engineering over the last 2 decades may offer alternative approaches for the urinary tract reconstruction. The main components of tissue engineering include biomaterials and cells. Biomaterials can be used with or without cultured cells. This paper focuses on cell sources, biomaterials, and existing methods of tissue engineering for urinary tract reconstruction in China. The paper also details challenges and perspectives involved in urinary tract reconstruction.
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Affiliation(s)
- Qingsong Zou
- Department of Urology, Affiliated Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, China
| | - Qiang Fu
- Department of Urology, Affiliated Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, China
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