Biophysical Cues on Modulation of Myoblast Differentiation Using Hydrostatic Pressure and Elastomeric Matrix of Tunable Stiffness

Accumulating evidence has demonstrated that integrated biochemical and biophysical microenvironmental cues with accurate temporal and spatial arrangements are needed in cell growth, self-renewal, and differentiation. Although muscle tissue pressure exists during the process of adult injury-induced myogenesis, a dearth of studies showed whether muscle regeneration would be altered when compressive pressure was applied during myoblast differentiation. Thus, the purpose of this study was to investigate the effects of hydrostatic pressure on differentiation of myoblasts, which was cultured on extracellular matrix of different stiffness to simulate the condition of embryonic myogenesis.
We cultured H9C2 myoblasts on fibronectin-coated polyacrylamide hydrogel, of which the stiffness was adjusted by different combinations of acrylamide and bis-acrylamide. Hydrostatic pressure of 10 cmH2O was applied. Then, we examined the expression of myogenesis transcriptional factors, MyoD and myogenin, using immunofluorescence staining. The results revealed that hydrostatic pressure increased the expression of MyoD and myogenin during myoblast differentiation. It might imply that hydrostatic pressure could facilitate the process of myogenesis. Therefore, further studies are merited on the detailed signaling pathways of pressure-induced modulation in myogenesis for developing muscle tissue engineering.