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This study explores the role of redox\u2010modulated surface thiol modification in myogenesis, using tris(2\u2010carboxyethyl)phosphine hydrochloride (TCEP) to reduce disulfide bonds and generate free thiol groups on the cell surface. The findings show that TCEP treatment significantly increases cell surface thiols, activating the focal adhesion kinase (FAK)\u2010phosphoinositide 3\u2010kinase (PI3K)\u2010protein kinase B (AKT) signaling pathway, and promoting cell spreading, adhesion, and actin cytoskeleton remodeling. This biochemical modulation upregulates myogenic and fusion\u2010related markers, facilitating multinucleated myotube formation. Biophysical analysis using traction force microscopy (TFM), intracellular force microscopy (IFM), and monolayer stress microscopy (MSM) demonstrates that TCEP\u2010treated cells, particularly at muscle\u2010relevant stiffness (20\u00a0kPa), exhibit increased traction, intracellular, and monolayer stresses compared to untreated cells. 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This proof\u2010of\u2010concept study advances the understanding of how redox\u2010modulated surface engineering and substrate stiffness regulate myogenesis, offering novel therapeutic strategies for muscle regeneration and optimization of stem cell\u2010based therapies for muscle tissue repair<\/jats:p>","DOI":"10.1002\/adhm.202500914","type":"journal-article","created":{"date-parts":[[2025,8,11]],"date-time":"2025-08-11T10:40:16Z","timestamp":1754908816000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Cell Surface Thiol Engineering Mechanoregulates Myogenic Differentiation via the FAK\u2013PI3K\u2013AKT Axis"],"prefix":"10.1002","volume":"15","author":[{"given":"Juyeon","family":"Kim","sequence":"first","affiliation":[{"name":"Soonchunghyang Institute of Medi\u2010Bio Science (SIMS) Soonchunhyang University  Cheonan\u2010si 31151 Republic of Korea"},{"name":"Department of Integrated Biomedical Science 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