Abstract
BACKGROUNS: Skeletal muscle stem cells (SkMSCs) are essential for muscle regeneration and represent a promising therapeutic target for muscle disorders. However, effective strategies to precisely regulate SkMSC fate by integrating biochemical and mechanical cues remain limited. METHODS: A decellularized extracellular matrix (ECM) hydrogel replicating the native muscle microenvironment was developed. The hydrogel was loaded with TGFβ1 and applied to SkMSCs in vitro and in Sprague-Dawley rat models. Signaling activation, m6A methylation of integrin mRNA, ERK phosphorylation, and functional outcomes were assessed through molecular and physiological analyses. RESULTS: The TGFβ1-loaded ECM hydrogel significantly enhanced SkMSC proliferation and differentiation by activating ERK signaling. Mechanistically, TGFβ1 promoted m6A methylation of integrin mRNA, leading to sustained ERK phosphorylation. In vivo, ECM hydrogels and TGFβ1 synergistically improved SkMSC function and muscle regeneration via m6A-modulated integrin signaling. CONCLUSION: This study reveals a novel m6A-mediated pathway that integrates biochemical (TGFβ1) and mechanical (ECM hydrogel) signals to direct SkMSC fate and provides a promising biomaterial-based strategy for treating muscle diseases through microenvironment-mimicking regenerative engineering.