title: The role of calcium signalling in the chondrogenic response of mesenchymal stem cells to hydrostatic pressure.
abstract: The object of this study was to elucidate the role of Ca++ signalling in the chondrogenic response of mesenchymal stem cells (MSCs) to hydrostatic pressure (HP). MSCs were seeded into agarose hydrogels, subjected to HP or kept in free swelling conditions, and were cultured either with or without pharmacological inhibitors of Ca++ mobility and downstream targets. Chelating free Ca++, inhibiting voltage-gated calcium channels, and depleting intracellular calcium storessuppressed the beneficial effect of HP on chondrogenesis, indicating that Ca++ mobility may play an important role in the mechanotransduction of HP. However, inhibition of stretch-activated calcium channels in the current experiment yielded similar results to the control group, suggesting that mechanotransduction of HP is distinct from loads that generate cell deformations. Inhibition of the downstream targets calmodulin, calmodulin kinase II, and calcineurin all knocked down the effect of HP on chondrogenesis, implicating these targets in MSCs response to HP. All of the pharmacological inhibitors that abolished the chondrogenic response to HP also maintained a punctate vimentin organisation in the presence of HP, as opposed to the mechanoresponsive groups where the vimentin structure became more diffuse. These results suggest that Ca++ signalling may transduce HP via vimentin adaptation to loading.
authors.affiliation: Department of Aerospace and Mechanical Engineering, 145 Multidisciplinary Research Building, University of Notre Dame, Notre Dame, IN 46556, USA.dwagner@nd.edu.
authors.forename: A J
authors.initials: AJ
authors.lastname: Steward
authors.forename: D J
authors.initials: DJ
authors.lastname: Kelly
authors.forename: D R
authors.initials: DR
authors.lastname: Wagner
mesh_terms: D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D065096:Calcium Chelating Agents / Q000494:pharmacology; D020013:Calcium Signaling; D002478:Cells, Cultured; D020219:Chondrogenesis; D006874:Hydrostatic Pressure; D040542:Mechanotransduction, Cellular; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D013552:Swine; D014746:Vimentin / Q000378:metabolism
chemical_list: D002121:Calcium Channel Blockers; D065096:Calcium Chelating Agents; D014746:Vimentin