Fate of tenogenic differentiation potential of human bone marrow stromal cells by uniaxial stretching affected by stretch-activated calcium channel agonist gadolinium

Nam, H.Y. and Raghavendran, H.R.B. and Pingguan-Murphy, Belinda and Abbas, A.A. and Merican, A.M. and Kamarul, Tunku (2017) Fate of tenogenic differentiation potential of human bone marrow stromal cells by uniaxial stretching affected by stretch-activated calcium channel agonist gadolinium. PLoS ONE, 12 (6). e0178117. ISSN 1932-6203

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Official URL: http://dx.doi.org/10.1371/journal.pone.0178117

Abstract

The role for mechanical stimulation in the control of cell fate has been previously proposed, suggesting that there may be a role of mechanical conditioning in directing mesenchymal stromal cells (MSCs) towards specific lineage for tissue engineering applications. Although previous studies have reported that calcium signalling is involved in regulating many cellular processes in many cell types, its role in managing cellular responses to tensile loading (mechanotransduction) of MSCs has not been fully elucidated. In order to establish this, we disrupted calcium signalling by blocking stretch-activated calcium channel (SACC) in human MSCs (hMSCs) in vitro. Passaged-2 hMSCs were exposed to cyclic tensile loading (1 Hz + 8% for 6, 24, 48, and 72 hours) in the presence of the SACC blocker, gadolinium. Analyses include image observations of immunochemistry and immunofluorescence staining from extracellular matrix (ECM) production, and measuring related tenogenic and apoptosis gene marker expression. Uniaxial tensile loading increased the expression of tenogenic markers and ECM production. However, exposure to strain in the presence of 20 μM gadolinium reduced the induction of almost all tenogenic markers and ECM staining, suggesting that SACC acts as a mechanosensor in strain-induced hMSC tenogenic differentiation process. Although cell death was observed in prolonged stretching, it did not appear to be apoptosis mediated. In conclusion, the knowledge gained in this study by elucidating the role of calcium in MSC mechanotransduction processes, and that in prolonged stretching results in non-apoptosis mediated cell death may be potential useful for regenerative medicine applications.

Item Type: Article
Uncontrolled Keywords: Aged; Apoptosis; Cadherins; Calcium Channel Agonists; Cell Differentiation; Cells, Cultured; Collagen; Extracellular Matrix; Fibronectins; Gadolinium; Humans; Mechanotransduction, Cellular; Mesenchymal Stromal Cells; Middle Aged; Stress, Mechanical; Tissue Engineering
Subjects: R Medicine
T Technology > T Technology (General)
Divisions: Faculty of Engineering
Faculty of Medicine
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 04 Sep 2018 04:04
Last Modified: 10 Oct 2018 09:12
URI: http://eprints.um.edu.my/id/eprint/19081

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