Diabetic ulcer is a prevalent complication affecting a large number of individuals globally and presents a significant challenge in healthcare, often resulting in prolonged healing times and an increased risk of infection. Conventional treatment options, such as proton pump inhibitors (PPIs) and H2 blockers, can cause adverse side effects. To address these limitations, we have developed a smart nanofiber patch incorporating silver nanoparticles (AgNPs) and glucose oxidase (GOx) for enhanced peptic ulcer wound healing. The AgNPs were embedded in PVA–alginate nanofibers to provide sustained antibacterial activity. In addition, nanofibers catalyse the oxidation of glucose in the wound exudate, generating hydrogen peroxide (H2O2), which enhances tissue regeneration and promotes wound healing. The characterization techniques such as SEM, FT-IR confirmed the nanofiber formation, and the release kinetics study demonstrated the sustained release of GOx from the nanofiber matrix. Further, in vitro studies demonstrated that the AgNPs-GOx nanofiber patch exhibited excellent cytocompatibility and enhanced cell proliferation. These findings highlight the promising potential of the AgNPs-GOx nanofiber patch as a novel therapeutic strategy for effective peptic ulcer treatment. In conclusion, the AgNPs-GOx nanofiber patch represents a significant advancement in peptic ulcer treatment, offering a promising alternative to conventional treatment options with fewer side effects. Its ability to provide sustained antibacterial activity and promote wound healing through a targeted and localized approach holds immense therapeutic potential.