This study aimed to fabricate a polyvinyl alcohol/chitosan (PVA/CS) nanofiber loaded with royal jelly (RJ) using blending electrospinning for potential wound dressings. The different PVA/CS ratios in electrospun nanofibers resulted in continuous nanofibers with an average diameter ranging from 219 to 299 nm. The FTIR spectra indicated that RJ was successfully incorporated into the nanofibers through hydrogen bonding with PVA/CS, which was further confirmed by the subsequent TGA experiments. Meanwhile, the RJ/PVA/CS nanofibers exhibited excellent water vapor permeability and hydrophilic properties. The encapsulation efficiency of RJ reached the maximum value of 89.00 %, while the cumulative release rate was up to 84.87 %. Furthermore, the RJ/PVA/CS nanofibers could inhibit the growth of Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli). The optimal PVA/CS ratio was determined to be 7:3, achieving inhibition rates of 97.83 % for S. aureus and 72.08 % for E. coli, demonstrating an excellent antibacterial performance. Therefore, this study successfully fabricated a wound dressing nanofiber with potential antibacterial efficacy.