Manufacturing, Cell Regulation, and Coculture Strategies for Vascularization and Neural Innervation of Skeletal Muscle Tissue

Skeletal muscle tissue engineering (SMTE) offers a promising approach for restoring volumetric muscle loss (VML), but recreating the anisotropic architecture, vascular networks, and neural structures of native muscle remains a major challenge. This review proposes that functional skeletal muscle reconstruction requires an integrated framework combining structural fabrication, cellular regulation, and multicellular coordination. For fabrication, it highlights the shift from single manufacturing methods to hybrid strategies, as single approaches cannot meet the multiscale structural requirements of skeletal muscle, while hybrid processes better reproduce native tissue features. For cellular regulation, the review emphasizes the roles of physical and chemical cues in promoting myogenic differentiation, vascularization, and innervation, noting that multimodal stimulation can synergistically mimic the dynamic muscle microenvironment and accelerate tissue maturation. For multicellular coculture, it identifies coculture systems as essential for functional remodeling and discusses related clinical translation and regulatory challenges. Future progress in SMTE will therefore rely on the convergence of advanced hybrid manufacturing strategies, controlled cellular regulation strategies, and robust multicellular coculture strategies, ultimately enabling the reproducible construction of vascularized and innervated muscle constructs with clinically relevant functionality.