A highly aligned ternary nanofiber matrix composed of poly(lactide-co-ε-caprolactone) (PLCL), collagen, and Ti₃C₂Tₓ MXene nanoparticles (PCM matrices) was developed to promote the regeneration of volumetric muscle loss (VML). The PCM matrices exhibited favorable physicochemical properties, structural uniformity, alignment, microporosity, and hydrophilicity, which supported fast regeneration of VML in vivo. The MXene nanoparticles activated inducible nitric oxide synthase and serum/glucocorticoid regulated kinase 1-mediated mTOR-AKT pathway, promoting myoblast differentiation and maturation. The study demonstrated that the PCM matrices enhanced muscle remodeling and recovery in mice following VML injury. Mechanistic insights from next-generation sequencing revealed that MXene NPs facilitated protein and ion availability within PCM matrices, leading to elevated intracellular Ca²⁺ levels in myoblasts through the activation of inducible nitric oxide synthase (iNOS) and serum/glucocorticoid regulated kinase 1 (SGK1), ultimately promoting myogenic differentiation via the mTOR-AKT pathway. Additionally, upregulated iNOS and increased NO⁻ contributed to myoblast proliferation and fiber fusion, facilitating overall myoblast maturation. These findings highlight the potential of MXene NPs loaded within highly aligned matrices as therapeutic agents to promote skeletal muscle tissue recovery. The PCM matrices were fabricated using electrospinning, with PLCL, collagen, and MXene NPs incorporated into the nanofibrous matrices. The physicochemical properties of the nanofibrous matrices were characterized, including fiber diameter, surface roughness, hydrophilicity, and mechanical properties. The PCM matrices showed enhanced cell adhesion, proliferation, and myogenic differentiation of C2C12 myoblasts compared to control matrices. Immunohistochemical analysis of mouse VML models showed that PCM matrices significantly enhanced muscle regeneration, with increased muscle mass, fiber diameter, and reduced inflammatory cell count. The study also identified key genes involved in myogenesis, including myostatin, MyoD, myogenin, and PGC-1α, which were upregulated in the PCM group. The results suggest that PCM matrices support the entire myogenic differentiation process in C2C12 cells, making them a promising approach for VML recovery. The study highlights the potential of MXene NPs in promoting spontaneous myogenesis through the activation of signaling pathways and the modulation of gene expression. The findings suggest that PCM matrices could be a promising clinical approach for VML recovery.A highly aligned ternary nanofiber matrix composed of poly(lactide-co-ε-caprolactone) (PLCL), collagen, and Ti₃C₂Tₓ MXene nanoparticles (PCM matrices) was developed to promote the regeneration of volumetric muscle loss (VML). The PCM matrices exhibited favorable physicochemical properties, structural uniformity, alignment, microporosity, and hydrophilicity, which supported fast regeneration of VML in vivo. The MXene nanoparticles activated inducible nitric oxide synthase and serum/glucocorticoid regulated kinase 1-mediated mTOR-AKT pathway, promoting myoblast differentiation and maturation. The study demonstrated that the PCM matrices enhanced muscle remodeling and recovery in mice following VML injury. Mechanistic insights from next-generation sequencing revealed that MXene NPs facilitated protein and ion availability within PCM matrices, leading to elevated intracellular Ca²⁺ levels in myoblasts through the activation of inducible nitric oxide synthase (iNOS) and serum/glucocorticoid regulated kinase 1 (SGK1), ultimately promoting myogenic differentiation via the mTOR-AKT pathway. Additionally, upregulated iNOS and increased NO⁻ contributed to myoblast proliferation and fiber fusion, facilitating overall myoblast maturation. These findings highlight the potential of MXene NPs loaded within highly aligned matrices as therapeutic agents to promote skeletal muscle tissue recovery. The PCM matrices were fabricated using electrospinning, with PLCL, collagen, and MXene NPs incorporated into the nanofibrous matrices. The physicochemical properties of the nanofibrous matrices were characterized, including fiber diameter, surface roughness, hydrophilicity, and mechanical properties. The PCM matrices showed enhanced cell adhesion, proliferation, and myogenic differentiation of C2C12 myoblasts compared to control matrices. Immunohistochemical analysis of mouse VML models showed that PCM matrices significantly enhanced muscle regeneration, with increased muscle mass, fiber diameter, and reduced inflammatory cell count. The study also identified key genes involved in myogenesis, including myostatin, MyoD, myogenin, and PGC-1α, which were upregulated in the PCM group. The results suggest that PCM matrices support the entire myogenic differentiation process in C2C12 cells, making them a promising approach for VML recovery. The study highlights the potential of MXene NPs in promoting spontaneous myogenesis through the activation of signaling pathways and the modulation of gene expression. The findings suggest that PCM matrices could be a promising clinical approach for VML recovery.