Mechanical Strain Promotes Proliferation of Adipose-Derived Stem Cells Through the Integrin β1-Mediated RhoA/Myosin Light Chain Pathway.

External volume expansion (EVE) promotes proliferation of adipose-derived stem cells (ADSCs) during adipose tissue regeneration. However, the mechanism by which EVE is translated into biochemical signals and subsequently induces proliferation of ADSCs is poorly understood. Here, we investigated the strain in adipose tissue and mechanochemical signaling upon EVE in rats. In addition, the effect of mechanical strain on proliferation of ADSCs was assessed using a custom-built Flexcell device. The level of strain in adipose tissue upon EVE peaked at week 1 and then decreased over time, and the cell proliferation rate was similarly affected. Mechanical strain-dependent activation of integrin β1 and the RhoA/myosin light chain (MLC) pathway was involved in cell proliferation. The proliferation rate of ADSCs was higher under 12% mechanical strain than under 6% and 0% mechanical strain in vitro. Mechanical strain-dependent activation of integrin β1 promoted activation of the small GTPase RhoA and phosphorylation of MLC. Furthermore, knockdown of integrin β1 attenuated activation of the RhoA/MLC pathway and proliferation of ADSCs in response to mechanical strain. Taken together, this study provides the first evidence of mechanochemical signaling in response to EVE. These data may help elucidate the effects of different strain levels on adipose tissue regeneration. Impact statement External volume expansion (EVE) induces adipose tissue regeneration and has great therapeutic potential to correct soft tissue defects. This study showed that EVE promotes proliferation of adipose-derived stem cells by activating integrin β1 and its crucial downstream signaling molecules, namely the small GTPase RhoA and p-myosin light chain. The findings of this study may assist clinical tissue engineering applications and provide new insights into the regulation of adipose tissue regeneration in clinical practice.