<p>Recent advancements in nanofiber-based biomaterials have demonstrated considerable potential in wound healing, attributed to their distinctive characteristics, including porosity, high surface area-to-volume ratio and the capacity to replicate the extracellular matrix. In this study polyvinyl alcohol (PVA), collagen and sodium alginate nanofibers were prepared and characterized for potential use in wound healing. The nanofiber’s chemical composition, morphology, thermal stability, mechanical strength, hydrophilicity and cell viability were examined using various characterization techniques. FTIR results confirmed the effective blending of polymer components, SEM analysis revealed the nanofibers are homogenous and interconnected morphology, Tensile testing verified suitable mechanical strength and flexibility, TGA showed sufficient thermal stability for biomedical usage. A contact angle of 39.51° signifies favorable hydrophilicity, proving that the Nano fibrous scaffold is appropriate for sustaining a moist wound environment. Excellent cytocompatibility was indicated by the high cell viability (98.8 ± 0.20%). Cell migration tests supported wound closure by showing intact cellular motility (94.3 ± 2.41%). DAPI labelling demonstrated a fluorescent intensity of 20.09 ± 0.70 a.u., showing preserved nuclear integrity, while AO/EtBr staining showed 16.53 ± 0.89% live cells and 3.21 ± 0.36% dead cells. Additionally, the Nano fibrous scaffold demonstrated antibacterial efficacy against <i>Escherichia coli</i>, and Staphylococcus aureus. Overall, the results show that the PVA/collagen/sodium alginate Nanofibrous scaffold is a viable material for wound dressings because it providesa well-balanced platform for tissue regeneration and infection control.</p> Graphical abstract <p></p>