This chapter deals with significant clinical challenges presented by musculoskeletal injuries and degenerative conditions, which frequently demand tissue replacement or repair procedures. Traditional biomaterials used in tissue engineering and regenerative medicine have demonstrated limitations in replicating the complexity of native tissues. The decellularized extracellular matrix (dECM) can provide a biologically active scaffold that closely resembles the natural tissue microenvironment, which has become a promising biomaterial as it still has structural proteins, signaling molecules, and vital biochemical indicators. The possible sources of dECM in musculoskeletal applications are discussed in this chapter, with a focus on the restoration of bone, cartilage, and tendon tissues. Also, deliberated are the various decellularization techniques, the significance of maintaining the integrity of the extracellular matrix, and methods to improve the mechanical and bioactive qualities of the matrix. Additionally, since these elements are essential for encouraging effective tissue regeneration, the relationship between dECM and stem cells as well as its function in regulating the immune response are highlighted. The effectiveness of dECM-based scaffolds in promoting tissue repair and functional recovery is demonstrated by preclinical research. Nonetheless, there are still issues to be resolved, including immunogenicity, matrix composition variability, and scalability for clinical use. In conclusion, dECM is a versatile and promising platform for musculoskeletal tissue engineering. It has the potential to improve clinical outcomes in orthopedic and sports medicine.