<p>Chitosan nanofibers, renowned for their unique properties, hold great promise for filtration, wound healing, and other biomedical applications. However, their instability in aqueous environments remains a major limitation, restricting practical implementation. This study introduces a scalable, non-toxic, UV-induced photocrosslinking method that overcomes this challenge while ensuring compatibility with continuous industrial production. To enable rapid photocrosslinking, chitosan was chemically modified with methacrylate groups using glycidyl methacrylate, facilitating efficient crosslinking upon UV exposure. Electrospinning a blend of methacrylated chitosan and poly(ethylene oxide), followed by photocrosslinking, produced nanofibers with significantly enhanced water stability, while maintaining biocompatibility with mammalian cells. Unlike conventional crosslinking techniques that rely on toxic reagents or prolonged thermal treatments, this approach eliminates hazardous chemicals and supports high-throughput, roll-to-roll manufacturing. Furthermore, we investigated the incorporation of gelatin, demonstrating its influence on nanofiber integrity and cell adhesion, which is crucial for biomedical applications. By integrating photocrosslinking into a streamlined industrial workflow developed with our industrial partner, this study removes a key scalability barrier in nanofiber production and offers a viable route to mass production of stable chitosan nanofibers.</p>