assisted extraction (USAE) and comprehensively characterized to evaluate their potential for biomedical applications. Extraction conditions were optimized through response surface methodology, yielding 8.93% (HWE) and 8.40% (USAE), confirming the high efficiency and reduced processing time of the ultrasound-assisted method. Although both extraction approaches produced polysaccharides with similar monosaccharide compositions, the USAE-derived sample demonstrated markedly superior quality, including the absence of protein (vs. 8.2% in HWE), lower ash content (4.3%), stronger antioxidant activity, higher apparent viscosity, and enhanced colloidal stability (–71 mV vs. –56 mV). These findings highlighted the USAE-derived polysaccharide as the optimal candidate for nanofiber fabrication. Pure polysaccharide solutions were not electrospinnable; however, blending with 6% (w/v) polyvinyl alcohol (PVA) enabled the formation of continuous, bead-free nanofibers at low polysaccharide concentrations (0.25–0.5% w/v). The resulting PVA/polysaccharide nanofibers exhibited high porosity, improved thermal stability, and enhanced structural integrity following glutaraldehyde vapor crosslinking. Swelling and degradation assessments further confirmed their suitability for wound dressing applications. Release modeling based on the free-volume theory successfully captured the non-linear, concentration-dependent diffusion behavior of the polysaccharide within the electrospun fibers. The 0.25% formulation achieved complete release within 72 hours, while the 0.5% formulation displayed slower and incomplete release due to reduced free volume, higher structural density, and greater fiber thickness, which collectively limited late-stage diffusion. In vitro cytocompatibility studies using L929 fibroblasts revealed that both formulations were non-cytotoxic; however, nanofibers containing 0.25% polysaccharide demonstrated superior cell viability and enhanced cell spreading and adhesion. This impro