Abstract In this study, the polyvinyl alcohol (PVA) and sodium caseinate (SC) nanofibers were produced by a single‐fluid electrospinning method from their blends. Afterward, the cross‐linking process with two different methods was applied to the PVA/SC (70/30, v/v) ratio, which was selected according to the surface and mechanical properties of the electrospun mat. In the first method, different ratios (15%, 20%, 25%, and 30%) of glutaraldehyde (GLA) cross‐linking agents were added to the PVA/SC solution and then, PVA/SC/GLA nanofibers were obtained. In the second method (in‐situ method), the nanofibers obtained from the PVA/SC solution were cross‐linked by dipping into the cross‐linking solution. After, PVA/SC/GLA/Zinc oxide nanoparticles (ZnO NP) mats were obtained by adding ZnO NP at different rates to the PVA/SC/GLA (7030‐25GLA) solution, which was chosen according to the results of thermal, mechanical, and moisture test. In addition, performing tests, a cytotoxicity test for fibroblast cell line (L929), and in vitro antibacterial test for Escherichia coli and Staphylococcus aureus were also applied to them. Therefore, the usability of PVA/SC/GLA/ZnO NP nanofibers as an antibacterial effective wound dressing was investigated. Due to the high toxic effect of GLA, it was found that PVA/SC/ZnO cross‐linked nanofibers are not suitable for wound dressing use. However, it was determined that the PVA/SC nanofiber cross‐linked by the in‐situ method had high cell viability according to the cytotoxicity test result and thus could be used as a fibroblast tissue scaffold.
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