This study explores the potential of electrospun polyvinyl alcohol (PVA) incorporating Eucalyptus camaldurensis extract (ELE) as an innovative wound dressing strategy to address the escalating threat of antibiotic resistance and associated complications in bacterial infections of wounds. The investigation is grounded in the recognition of the antibacterial properties inherent in medicinal plants and the advantageous release characteristics of nanomaterials, particularly electrospun nanofibers that closely mimic the extracellular matrix. Utilizing the electrospinning technique, nanofiber mats were fabricated with hydroalcoholic Eucalyptus camaldulensis extract, and their structural and morphological attributes were comprehensively characterized using scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD). The study employed 60 male Wistar rats, categorizing them into groups treated with PVA/ELE, nitrofurazone, normal saline, and PVA wound dressings. Microbial and histopathological analyses were conducted at specified intervals post-infection. The results unveiled the remarkable antibacterial efficacy of PVA/ELE, as evidenced by a substantial reduction in bacterial count compared to control groups. Furthermore, the PVA/ELE group demonstrated superior wound size reduction, re-epithelization, and collagen deposition, akin to the effects observed in the nitrofurazone group. The findings suggest that PVA/ELE exhibits significant antimicrobial potential and promotes advanced wound-healing processes. Consequently, this electrospun nanofiber formulation, enriched with ELE, emerges as a promising and viable alternative for conventional wound care, offering multifaceted benefits in combating bacterial infections and facilitating wound healing.