Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, C039

Oral Communications

Skeletal myogenesis can be driven by key growth factors contained in human platelets.

D. Scully1, P. Sfyri1, S. Verpoorten1, A. Aburima1, K. naseem3, P. papadopoulos2, L. Gutierrez4, A. Matsakas1

1. Centre for Atherothrombosis & Metabolic Diseases, Hull York Medical School, Hull, East Riding of Yorkshire, United Kingdom. 2. Department of Hematology, Instituto de Investigación Sanitaria San Carlos (IdISSC), Hospital Clínico, Madrid, San Carlos, Spain. 3. Leeds Institute of Cardiovascular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom. 4. Dept. of Medicine, Universidad de Oviedo, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Asturias, Spain.

Introduction: Platelet-based applications offer a potentially straightforward, cost-effective and safe, autologous method of delivering growth factors to injured tissue, in an attempt to promote regeneration. However, there is currently contrasting evidence on the role of platelets as biomaterials in skeletal muscle injuries. Given that the molecular mechanisms involved in platelet-mediated regeneration are not well established, the aim of this study was to determine the effect of platelet releasate on skeletal myogenesis in vitro and muscle stem cells ex vivo. Methods: We studied the effect of platelet releasate on proliferation and differentiation of C2C12 myoblasts, single muscle fibres and isolated satellite cells by means of cell proliferation assays, immunohistochemistry and gene expression. Moreover, we expanded in vitro cell culture findings on by establishing the effect of human platelet releasate on murine skeletal muscle stem cells using protein expression profiles for key myogenic regulatory factors. Results: We show that platelet releasate increases myoblast proliferation in a dose-dependent manner, The proliferative effects were prevented by pharmacological inhibition of vascular endothelial growth factor- (VEGF) and/or platelet-derived growth factor- (PDGF) receptors. The cell culture findings were validated with primary skeletal muscle stem cells, where we observed that platelet releasate upregulated MyoD and Scrib expression on and promoted their myogenic potential through the VEGFR and PDGFR-mediated pathways. Application of platelet releasate can either attenuate fusion of myoblasts into myotubes or promote terminal myogenic differentiation in a temporal manner, indicating that the timing of releasate application is crucial. Key components of the platelet releasate were identified by immuno-based assays. Conclusion: Taken together, in vitro and ex vivo data provide supporting evidence of a positive effect of platelet releasate on myoblast proliferation, with temporal effects on cell differentiation. The effects are mediated through the VEGF and PDGF signalling pathways. Platelet-based methodologies aiming to provide autologous biomaterials have implications for regenerative medicine and muscle healing.

Where applicable, experiments conform with Society ethical requirements