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Peng L, Wu F, Cao M, Li M, Cui J, Liu L, Zhao Y, Yang J. Effects of different physical factors on osteogenic differentiation. Biochimie 2023; 207:62-74. [PMID: 36336107 DOI: 10.1016/j.biochi.2022.10.020] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2022] [Revised: 10/11/2022] [Accepted: 10/31/2022] [Indexed: 11/06/2022]
Abstract
Osteoblasts are essential for bone formation and can perceive external mechanical stimuli, which are translated into biochemical responses that ultimately alter cell phenotypes and respond to environmental stimuli, described as mechanical transduction. These cells actively participate in osteogenesis and the formation and mineralisation of the extracellular bone matrix. This review summarises the basic physiological and biological mechanisms of five different physical stimuli, i.e. light, electricity, magnetism, force and sound, to induce osteogenesis; further, it summarises the effects of changing culture conditions on the morphology, structure and function of osteoblasts. These findings may provide a theoretical basis for further studies on bone physiology and pathology at the cytological level and will be useful in the clinical application of bone formation and bone regeneration technology.
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Affiliation(s)
- Li Peng
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China; Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, China
| | - Fanzi Wu
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China
| | - Mengjiao Cao
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China
| | - Mengxin Li
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China
| | - Jingyao Cui
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China
| | - Lijia Liu
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China
| | - Yun Zhao
- Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, China.
| | - Jing Yang
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, China.
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Marciniak J, Lossdörfer S, Knaup I, Bastian A, Craveiro RB, Jäger A, Wolf M. Orthodontic cell stress modifies proinflammatory cytokine expression in human PDL cells and induces immunomodulatory effects via TLR-4 signaling in vitro. Clin Oral Investig 2019; 24:1411-1419. [PMID: 31691860 DOI: 10.1007/s00784-019-03111-8] [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] [Received: 02/20/2019] [Accepted: 10/07/2019] [Indexed: 12/17/2022]
Abstract
OBJECTIVE Biomechanical orthodontics loading of the periodontium initiates a cascade of inflammatory signaling events that induce periodontal remodeling and finally facilitate orthodontic tooth movement. Pattern recognition receptors such as toll-like receptors (TLRs) have been well characterized for their ability to induce the activation of inflammatory, immunomodulatory cytokines. Here, we examined whether the cellular response of human periodontal ligament (hPDL) cells to mechanical stress involves TLR-4 signaling in vitro. MATERIALS AND METHODS Confluent hPDL cells were cultured in the presence of 5 μg/ml TLR-4 antibody (TLR-4ab) for 1 h prior to the induction of compressive forces by the use of round glass plates for 24 h. At harvest, interleukin-6 and interleukin-8 (IL-6, IL-8) mRNA and protein expression were analyzed by real-time PCR and ELISA. The immunomodulatory role of mechanical cell stress and TLR-4 signaling was addressed in co-culture experiments of hPDL and THP-1 cells targeting monocyte adhesion and by culturing osteoclastic precursors (RAW 264.7) in the presence of the conditioned medium of hPDL cells that had been mechanically loaded before. RESULTS Basal expression of IL-6 and IL-8 was not affected by TLR-4ab, but increased significantly upon mechanical loading of hPDL cells. When cells were mechanically stressed in the presence of TLR-4ab, the effect seen for loading alone was markedly reduced. Likewise, monocyte adhesion and osteoclastic differentiation were enhanced significantly by mechanical stress of hPDL cells and this effect was partially inhibited by TLR-4ab. CONCLUSIONS The results of the present study indicate a proinflammatory and immunomodulatory influence of mechanical loading on hPDL cells. Intracellular signaling involves a TLR-4-dependent pathway. CLINICAL RELEVANCE These findings hold out the prospect of interfering with the cellular response to mechanical cell stress in order to minimize undesired side effects of orthodontic tooth movement.
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Affiliation(s)
- Jana Marciniak
- Department of Orthodontics, Dental Clinic, University of Aachen, Pauwelsstr. 30, 52074, Aachen, Germany.,Department of Orthodontics, Dental Clinic, University of Bonn, Bonn, Germany
| | - Stefan Lossdörfer
- Department of Orthodontics, Dental Clinic, University of Bonn, Bonn, Germany
| | - Isabel Knaup
- Department of Orthodontics, Dental Clinic, University of Aachen, Pauwelsstr. 30, 52074, Aachen, Germany
| | - Asisa Bastian
- Department of Orthodontics, Dental Clinic, University of Aachen, Pauwelsstr. 30, 52074, Aachen, Germany
| | - Rogerio B Craveiro
- Department of Orthodontics, Dental Clinic, University of Aachen, Pauwelsstr. 30, 52074, Aachen, Germany
| | - Andreas Jäger
- Department of Orthodontics, Dental Clinic, University of Bonn, Bonn, Germany
| | - Michael Wolf
- Department of Orthodontics, Dental Clinic, University of Aachen, Pauwelsstr. 30, 52074, Aachen, Germany.
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Xu W, Lu Y, Yue J, Lu W, Zhou W, Zhou X, Ye L, Zheng Q, Zhang L, Huang D. Occlusal trauma inhibits osteoblast differentiation and bone formation through IKK-NF-κB signaling. J Periodontol 2019; 91:683-692. [PMID: 31487049 DOI: 10.1002/jper.18-0710] [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] [Received: 12/08/2018] [Revised: 07/22/2019] [Accepted: 07/30/2019] [Indexed: 02/05/2023]
Abstract
BACKGROUND Occlusal trauma is an important factor promoting bone loss caused by periodontal diseases. Although there are reports of traumatic force promoting bone resorption in periodontal diseases, no studies examining the inhibition of bone formation by traumatic force and the underlying mechanism have been reported. The aim of this study was to investigate the mechanism whereby traumatic force inhibits bone formation. METHODS MC3T3-E1 cells were induced to undergo osteogenic differentiation and subjected to cyclic uniaxial compressive stress with or without stimulation with Pg. LPS. The expression of osteoblast markers and the activation of IKK-NF-κB signaling were evaluated in vitro. Then, MC3T3-E1 cells were induced to undergo osteogenic differentiation and subjected to cyclic uniaxial compressive stress with or without IKK-2 Inhibitor VI. The expression of osteoblast markers was determined. Then, the classic Wnt signaling pathway (β-catenin, Gsk3β, p-Gsk3β, and Dkk1) was further evaluated in vitro. Finally, occlusal trauma was induced in Wistar rats with or without the injection of IKK-2 Inhibitor VI, to evaluate changes in bone mass and IKK-NF-κB and Wnt/β-catenin signaling in vivo. RESULTS After stimulation with Pg. LPS and traumatic force, IKK-NF-κB signaling was significantly activated in vitro. The expression of osteoblast markers and the activity of alkaline phosphatase in MC3T3-E1 cells declined after traumatic force loading and were rescued when IKK-NF-κB signaling was blocked. Wnt/β-catenin signaling was accordingly inhibited upon force loading, but this inhibition was reversed when IKK-NF-κB was antagonized in vitro. X-ray and Micro-CT analysis of the mandibles of the rats as well as HE and TRAP staining showed that bone loss induced by occlusal trauma declined after IKK-NF-κB was inhibited. The expression of p65 and IκBα was increased when occlusal trauma was induced in Wistar rats, whereas β-catenin, OCN, and Runx2 levels were decreased. After blocking IKK-NF-κB, significant upregulation of β-catenin, OCN, and Runx2 was observed in rats suffering from occlusal trauma. CONCLUSIONS IKK-NF-κB signaling could be activated by traumatic force or occlusal trauma. Its activation promoted the degradation of β-catenin, ultimately inhibiting osteogenic differentiation in vitro and bone formation in vivo.
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Affiliation(s)
- Weizhe Xu
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Ying Lu
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.,Department of Conservative Dentistry, College of Stomatology, Dalian Medical University, Dalian, Liaoning, China
| | - Junli Yue
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Wanlu Lu
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Wei Zhou
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Xuedong Zhou
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Ling Ye
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Qinghua Zheng
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Lan Zhang
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
| | - Dingming Huang
- State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Cariology and Endodontics West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China
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Yao Y, Mak AF. Strengthening of C2C12 mouse myoblasts against compression damage by mild cyclic compressive stimulation. J Biomech 2016; 49:3956-3961. [PMID: 27884430 DOI: 10.1016/j.jbiomech.2016.11.050] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/17/2016] [Revised: 11/10/2016] [Accepted: 11/11/2016] [Indexed: 11/28/2022]
Abstract
Deep tissue injury (DTI) is a severe kind of pressure ulcers formed by sustained deformation of muscle tissues over bony prominences. As a major clinical issue, DTI affects people with physical disabilities, and is obviously related to the load-bearing capacity of muscle cells in various in-vivo conditions. It is important to provide a preventive approach to help muscle cells from being damaged by compressive stress. In this study, we hypothesized that cyclic compressive stimulation could strengthen muscle cells against compressive damage and enhance the cell plasma membrane resealing capability. Monolayer of myoblasts was cultured in the cell culture dish covered by a cylinder 0.5% agarose gel. The platen indenter was applied with 20% strain on the agarose gel in the Mach-1 micromechanical system. The vibration was 1Hz sinusoidal function with amplitude 0.2% strain based on 20% gel strain. Cyclic compressive stimulation for 2h could enhance the compressive stress damage threshold of muscle cells, the muscle cell plasma membrane resealing ratio and viability of muscle cell under static loading as preventive approach. This approach might help to reduce the risk of DTI in clinic.
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Affiliation(s)
- Yifei Yao
- Division of Biomedical Engineering, The Chinese University of Hong Kong, Hong Kong Special Administrative Region
| | - Arthur Ft Mak
- Division of Biomedical Engineering, The Chinese University of Hong Kong, Hong Kong Special Administrative Region.
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Lu Y, Zheng Q, Lu W, Yue J, Zhou W, Zhou X, Zhang L, Huang D. Compressive mechanical stress may activate IKK-NF-κB through proinflammatory cytokines in MC3T3-E1 cells. Biotechnol Lett 2015; 37:1729-35. [DOI: 10.1007/s10529-015-1849-6] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/09/2015] [Accepted: 05/01/2015] [Indexed: 01/13/2023]
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Basson MD, Zeng B, Downey C, Sirivelu MP, Tepe JJ. Increased extracellular pressure stimulates tumor proliferation by a mechanosensitive calcium channel and PKC-β. Mol Oncol 2015; 9:513-526. [PMID: 25454347 PMCID: PMC4487881 DOI: 10.1016/j.molonc.2014.10.008] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/02/2014] [Revised: 10/17/2014] [Accepted: 10/20/2014] [Indexed: 01/31/2023] Open
Abstract
Large tumors exhibit high interstitial pressure heightened by growth against the constraining stroma. Such pressures could stimulate tumor proliferation via a mechanosensitive ion channel. We studied the effects of 0-80 mmHg increased extracellular pressure for 24 h on proliferation of SW620, Caco-2, and CT-26 colon; MCF-7 breast; and MLL and PC3 prostate cancer cells, and delineated its mechanism in SW620 cells with specific inhibitors and siRNA. Finally, we compared NF-kB, phospho-IkB and cyclin D1 immunoreactivity in the high pressure centers and low pressure peripheries of human tumors. Pressure-stimulated proliferation in all cells. Pressure-driven SW620 proliferation required calcium influx via the T-type Ca(2+) channel Cav3.3, which stimulated PKC-β to invoke the IKK-IkB-NF-kB pathway to increase proliferation and S-phase fraction. The mitotic index and immunoreactivity of NF-kB, phospho-IkB, and cyclin D1 in the center of 28 large human colon, lung, and head and neck tumors exceeded that in tumor peripheries. Extracellular pressure increases [Ca(2+)]i via Cav3.3, driving a PKC-β- IKK- IkB-NF-kB pathway that stimulates cancer cell proliferation. Rapid proliferation in large stiff tumors may increase intratumoral pressure, activating this pathway to stimulate further proliferation in a feedback cycle that potentiates tumor growth. Targeting this pathway may inhibit proliferation in large unresectable tumors.
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Affiliation(s)
- Marc D Basson
- Department of Surgery, Michigan State University College of Human Medicine, 1200 E Michigan Ave, Lansing Charter Township, MI 48912, USA.
| | - Bixi Zeng
- Department of Surgery, Michigan State University College of Human Medicine, 1200 E Michigan Ave, Lansing Charter Township, MI 48912, USA
| | - Christina Downey
- Department of Surgery, Michigan State University College of Human Medicine, 1200 E Michigan Ave, Lansing Charter Township, MI 48912, USA
| | - Madhu P Sirivelu
- Department of Surgery, Michigan State University College of Human Medicine, 1200 E Michigan Ave, Lansing Charter Township, MI 48912, USA
| | - Jetze J Tepe
- Department of Pharmacology, Michigan State University, 1355 Bogue Street, B440 Life Sciences Building, East Lansing, MI 48824, USA
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Spyropoulou A, Basdra EK. Mechanotransduction in bone: Intervening in health and disease. World J Exp Med 2013; 3:74-86. [DOI: 10.5493/wjem.v3.i4.74] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/28/2013] [Revised: 09/06/2013] [Accepted: 11/03/2013] [Indexed: 02/06/2023] Open
Abstract
Mechanotransduction has been proven to be one of the most significant variables in bone remodeling and its alterations have been shown to result in a variety of bone diseases. Osteoporosis, Paget’s disease, orthopedic disorders, osteopetrosis as well as hyperparathyroidism and hyperthyroidism all comprise conditions which have been linked with deregulated bone remodeling. Although the significance of mechanotransduction for bone health and disease is unquestionable, the mechanisms behind this important process have not been fully understood. This review will discuss the molecules that have been found to be implicated in mechanotransduction, as well as the mechanisms underlying bone health and disease, emphasizing on what is already known as well as new molecules potentially taking part in conveying mechanical signals from the cell surface towards the nucleus under physiological or pathologic conditions. It will also focus on the model systems currently used in mechanotransduction studies, like osteoblast-like cells as well as three-dimensional constructs and their applications among others. It will also examine the role of mechanostimulatory techniques in preventing and treating bone degenerative diseases and consider their applications in osteoporosis, craniofacial development, skeletal deregulations, fracture treatment, neurologic injuries following stroke or spinal cord injury, dentistry, hearing problems and bone implant integration in the near future.
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Zhou S, Zhang J, Zheng H, Zhou Y, Chen F, Lin J. Inhibition of mechanical stress-induced NF-κB promotes bone formation. Oral Dis 2012; 19:59-64. [DOI: 10.1111/j.1601-0825.2012.01949.x] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
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Kook SH, Jang YS, Lee JC. Involvement of JNK-AP-1 and ERK-NF-κB signaling in tension-stimulated expression of Type I collagen and MMP-1 in human periodontal ligament fibroblasts. J Appl Physiol (1985) 2011; 111:1575-83. [DOI: 10.1152/japplphysiol.00348.2011] [Citation(s) in RCA: 59] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022] Open
Abstract
Type I collagen (COL I) and matrix metalloproteinase-1 (MMP-1) are the predominant matrix proteins in the extracellular matrix of the human periodontal ligament (PDL). The expression of these proteins in PDL fibroblasts (PLF) is sensitive to physiological and mechanical stress and is critical for PDL remodeling accompanied by alveolar bone remodeling. This study examined how dose tensile force regulates the expression of COL I and MMP-1 and explored the possible roles of mitogen-activated protein kinases (MAPKs) and transcription factors, such as activator protein-1 (AP-1) and nuclear factor-κB (NF-κB). Tensile force stimulated the mRNA expression of COL I and MMP-1 in the cells and also activated MAPKs including extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38 MAPK. A pharmacological inhibitor of ERK or JNK prevented the expression of matrix genes and the nuclear translocation of c-Jun proteins in the force-applied PLF. The knockdown of c-Jun by transfecting the cells with its antisense oligonucleotides reduced the force-induced increase in matrix gene expression. In particular, the ERK inhibitor but not JNK or p38 MAPK inhibitor attenuated the force-mediated stimulation of NF-κB-DNA binding and MMP-1 expression. Overall, these results highlight the mechanotransduction pathways involved in matrix gene expression in PLF, where the tension-stimulated expression of COL I and MMP-1 is controlled by the ERK/JNK-AP-1 and ERK-NF-κB signaling pathways.
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Affiliation(s)
- Sung-Ho Kook
- Division of Hematology and Oncology, Department of Medicine, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania
| | - Yong-Suk Jang
- Research Center of Bioactive Materials, Chonbuk National University
| | - Jeong-Chae Lee
- Research Center of Bioactive Materials, Chonbuk National University
- Cluster for Craniofacial Development and Regeneration Research, Institute of Oral Biosciences and School of Dentistry (BK21 Program), Chonbuk National University, Jeonju, South Korea
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Morris HL, Reed CI, Haycock JW, Reilly GC. Mechanisms of fluid-flow-induced matrix production in bone tissue engineering. Proc Inst Mech Eng H 2010; 224:1509-21. [DOI: 10.1243/09544119jeim751] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022]
Abstract
Matrix production by tissue-engineered bone is enhanced when the growing tissue is subjected to mechanical forces and/or fluid flow in bioreactor culture. Cells deposit collagen and mineral, depending upon the mechanical loading that they receive. However, the molecular mechanisms of flow-induced signal transduction in bone are poorly understood. The hyaluronan (HA) glycocalyx has been proposed as a potential mediator of mechanical forces in bone. Using a parallel-plate flow chamber the effects of removal of HA on flow-induced collagen production and NF-κB activation in MLO-A5 osteoid osteocytes were investigated. Short periods of fluid flow significantly increased collagen production and induced translocation of the NF-κB subunit p65 to the cell's nuclei in 65 per cent of the cell population. Enzymatic removal of the HA coat and antibody blocking of CD44 (a transmembrane protein that binds to HA) eliminated the fluid-flow-induced increase in collagen production but had no effect on the translocation of p65. HA and CD44 appear to play roles in transducing the flow signals that modulate collagen production over long-term culture but not in the short-term flow-induced activation of NF-κB, implying that multiple signalling events are initiated from the commencement of flow. Understanding the mechanotransduction events that enable fluid flow to stimulate bone matrix production will allow the optimization of bioreactor design and flow profiles for bone tissue engineering.
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Affiliation(s)
- H L Morris
- Department of Engineering Materials, Kroto Research Institute, University of Sheffield, Broad Lane, UK
| | - C I Reed
- Department of Engineering Materials, Kroto Research Institute, University of Sheffield, Broad Lane, UK
| | - J W Haycock
- Department of Engineering Materials, Kroto Research Institute, University of Sheffield, Broad Lane, UK
| | - G C Reilly
- Department of Engineering Materials, Kroto Research Institute, University of Sheffield, Broad Lane, UK
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Proteomic analysis of early-response to mechanical stress in neonatal rat mandibular condylar chondrocytes. J Cell Physiol 2010; 223:610-22. [DOI: 10.1002/jcp.22052] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
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Wang Y, Zhao Z, Li Y, Li Y, Wu J, Fan X, Yang P. Up-regulated alpha-actin expression is associated with cell adhesion ability in 3-D cultured myocytes subjected to mechanical stimulation. Mol Cell Biochem 2009; 338:175-81. [PMID: 20024607 DOI: 10.1007/s11010-009-0351-7] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2009] [Accepted: 12/03/2009] [Indexed: 12/15/2022]
Abstract
This study was aimed to investigate the alteration of alpha-actin in three-dimensionally (3-D) cultured myocytes under cyclic tensile stress loading. Myocytes were collected from neonatal SD rat's lateral pterygoid muscle for primary cell culture. The third-passage cells were implanted and 3-D cultured in poly lactic-co-glycolic acid (PLGA) scaffold, and then subjected to cyclic tensile stress (0.5 Hz, 2,000 microstrain) for 0, 2, 4, 8, 12, and 24 h through a four-point bending strain system. The alpha-actin mRNA was investigated by semi-quantitative RT-PCR. The alpha-actin protein expression was examined by immunofluorescent cytochemistry, laser confocal scanning microscopy (LCSM), and image analysis technology. The dynamic adhesion of myocytes to PLGA scaffolds was investigated by fluorescence microscope and the viability of the myocytes was measured by MTT assay. After mechanical loading, the alpha-actin mRNA increased at 2 h and then declined. The alpha-actin protein expression kept increased until peaked at 12 h, but declined at 24 h. The time course changing of alpha-actin protein expression parallelled with that of cell adhesion ability. It is concluded that alpha-actin expression is probably associated with cell adhesion ability in myocytes subjected to mechanical stimulation.
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Affiliation(s)
- Yu Wang
- Department of Orthodontics, State Key Laboratory of Oral Diseases, West China Stomatology Hospital Sichuan University, Chengdu, Sichuan, People's Republic of China
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