Mechanochemistry-Induced Universal Hydrogel Surface Modification for Orientation and Enhanced Differentiation of Skeletal Muscle Myoblasts

Micropatterned surface substrates containing topographic cues offer the possibility of programming tissue organization as a cell template by guiding cell alignment, adhesion, and function. In this study, we developed and used a force stamp method to grow aligned micropatterns with tunable chemical properties and elasticity on the surface of hydrogels based on a force-triggered polymerization mechanism of double-network hydrogels to elucidate the underlying mechanisms by which cells sense and respond to their mechanical and chemical microenvironments. In this work, we describe the impact of aligned micropatterns on the combined effects of microstructural chemistry and mechanics on the selective adhesion, directed migration, and differentiation of myoblasts. Our investigations revealed that topographically engineered substrates with hydrophobic and elevated surface roughness significantly enhanced myoblast adhesion kinetics. Concurrently, spatially ordered architectures facilitated cytoskeletal reorganization in myocytes, establishing biomechanically favorable niches for syncytial myotube development through extracellular matrix (ECM) physical guidance. Reverse transcription PCR analysis and immunofluorescence revealed that the expression of differentiation-specific genes, myosin heavy chain, and myogenic regulatory factors <i>Myf5</i> and <i>MyoD</i> was upregulated in muscle cells on the aligned patterned scaffolds. These results suggest that the aligned micropatterns can promote muscle cell differentiation, making them potential scaffolds for enhancing skeletal differentiation.