Bio-composite materials were fabricated for wound healing in three concentrations.• Biodegradable polymers were used to create electrospun fibers. • A blend of polyvinyl alcohol and chitosan, incorporated with nanoparticles, was used to modify mechanical properties.This study uses electro-spinning techniques to create a poly(vinyl alcohol)chitosan-zinc oxide nanoparticles (PVA+CS)+ZnO NPs system in three concentrations of ZnO for application as composite nanofibers for wound treatment applications and Employing the Taguchi technique to optimize the mechanical characteristics via a four-level experimental design process (L16).Numerous techniques have been utilized to characterize the nanofibrous scaffolds: contact angle measurement, cytotoxicity testing, proliferation testing, evaluation of antimicrobial activity, Fourier Transform Infrared Spectroscopy (FTIR), Field Emission Scanning Electron Microscopy (FESEM), Energy Dispersion X-ray Spectroscopy (EDX), and a comparative experimental study involving the determination of hardness using the Nanoindentation method for (PVA+CS)+ZnO. The ideal combination for adding ZnO as nanoparticles was found to be (PVA+CS)+0.6 ZnO, with a flow rate of 0.5 mL/hr, an applied voltage of 18 kV, and a needle tip-to-collector distance of 15 cm (position). This resulted in the smallest fiber diameter (48 nm) with a smooth and uniform distribution. As a consequence, (PVA+CS)+ZnO can be regarded as non-toxic in accordance with the criteria. The methyl thiazole tetrazolium (MTT) assay was used to evaluate the cell viability of L929 (cell cultures for skin). It significantly affects cell viability, achieving 50% in less than 24 hours, which means cell growth through 24 hours is necessary for embryonic development, tissue repair, and regulation of cell division in case of wound healing. Exhibiting findings of inhibition zone diameter in antibacterial activity test exceeding 20 mm. According to the results, (PVA+CS)+0.6 ZnO's hydrophilic properties showed a well-connected porous structure and made it easier for nutrients and oxygen to be exchanged, encouraging cellular activity. After evaluating the mechanical characteristics of each specimen, the main determinants of these characteristics are determined using a Taguchi orthogonal array L16 design. The results of this study suggest that (PVA+CS)+0.6 ZnO could be a good biomedical material for skin tissue engineering applications.