Rhamnus prinoides leaf contains carbohydrates, saccharides, phenolic acids, and diterpenes with antibacterial, wound-healing, and anti-inflammatory properties. In this study, Rhamnus prinoides leaf extract was successfully incorporated into polycaprolactone-cellulose acetate (PCL-CA) nanofibers through electrospinning technique for the first time. The mats' morphology, diameter, chemical, and crystalline structure were characterized. The study investigated the mats' antibacterial activity, wound healing, cytotoxicity, drug release behaviour, hydrophilicity, and water absorbency properties. The results revealed that the mats exhibited continuous, smooth, without-beads, and interconnected structures, with average fiber diameters ranging from 385 ± 21 nm to 332 ± 74 nm. The antibacterial effeciency was remarkable against S. aureus and E. coli, achieving bacterial reduction percentages exceeding 99 % at concentrations of 3 % and above against S. aureus and 5 % and above against E. coli. Cytotoxic tests showed low-cytotoxicity up to an extract concentration of 7 %. The extract release increases with an increase in concentration. In vitro wound healing assay, the mats enhanced cell migration to the wound area. Additionally, the incorporation of Rhamnus prinoides significantly improved the hydrophilicity and water absorbency of the nanofibers. Overall, the study highlights the mats' broad antimicrobial and wound healing properties with less cytotoxicity, hydrophilicity, and water absorbency, making them promising for use as wound dressings.