Engineering fibrous scaffolds with controllable structural and mechanical attributes is critical for reproducing the anisotropic organization and functional microenvironment of skeletal muscle. In this study, we developed gelatin-chitosan (GC) scaffolds using a toxic solvent-free tensile spinning process that, resulting in aligned fiber assemblies. Adjusting the chitosan fraction systematically altered the viscoelastic properties of the precursor mixture, which directly influenced fiber formation, pore characteristics (20-26%), hydration resilience, and stiffness (3-87 kPa). Spectroscopic and diffraction analyses revealed strengthened molecular interactions and increased structural order with chitosan incorporation. In parallel, surface charge measurements confirmed compositional regulation of electrostatic cues governing cell-matrix interactions.

Among the tested formulations, mid-range compositions (GC_1.00-GC_1.75) achieved an advantageous combination of limited swelling, persistent mass transport, and stable mechanical behavior under hydrated conditions. In particular, GC _1.25 scaffolds promoted robust myoblast attachment, high viability, and pronounced alignment of the actin cytoskeleton, driving directional myotube development along the fiber axis. Collectively, these results highlight gelatin-chitosan composites as a versatile system for dissecting how polysaccharide-protein interactions and rheological tuning dictate scaffold architecture, charge environment, and downstream myogenic outcomes. The approach outlined here offers a practical foundation for creating architecturally defined fibrous scaffolds capable of sustaining cellular organization and function.