Abstract: Objective To explore the feasibility of inducing myofibroblasts differentiation of skin fibroblasts by mechanical loading based on three dimensional culture in vitro. Methods Human dermal fibroblasts were isolated and three dimensionally cultured in type I rat tail collagen scaffold. Then mechanical loading were added by stretching the gel (stretch group, n=9), and control group was set up (non-stretch group, n=9). The situation of myofibroblasts differentiation were tested by drawing cell growth curve, immunofluorescent staining and Real-Time PCR. Results Cell proliferation was slowed down under mechanical loading environment. The cell proliferation was promoted in stretch group, but there was no significant difference comparing with non-stretch group. Dyeing brightness of α-SMA and fibronectin were significantly higher in stretch group showed by immunofluorescent staining. Higher mRNA expression of α-SMA, fibronectin, type I and III collagen genes were observed in stretch group by real time PCR. Conclusion The model of mechanical loading inducing myofibroblasts differentiation of skin fibroblasts based on 3D culture could be successfully established.

Key words: Myofibroblasts, Mechanical loading, 3D culture