In this study, electrospun PVA membranes crosslinked with citric acid or glutaraldehyde and containing the antibiotic chloramphenicol (CLF) were obtained for the production of dressings. This material was characterized by SEM, FTIR, DSC, TGA NMR, degree of swelling, and solubility. The SEM images showed the possibility of obtaining nanofibers with a diameter between 600 and 700 nm. Crosslinking with citric acid (3% m/m) was the only one able to give stability to PVA fibers in water and preserve their morphology. The stability of PVA fibers in water was associated with a decrease in OH groups in PVA and an increase in crystallinity, both effects promoted by crosslinking. The increase in crystallinity due to crosslinking was observed by DSC, DRX, and FTIR analyses. A linear correlation was found between the increase in enthalpy of fusion and the increase in crystallinity. The FTIR analysis confirmed the incorporation of the drug into the fibers. The results of TGA analysis suggested that the presence of basic groups in the CLF molecule may act as proton receptors delaying the first stage of thermal degradation of the polymer. The absence of CLF-related peaks in the X-ray diffractograms and DSC curves suggests that the drug may be in an amorphous state, which improves the solubility of CLF and facilitates its immediate release, making the drug more bioavailable. The mechanism of CLF release indicated that the kinetics follows the typical Weibull model for matrix systems, showing an immediate release in the first 30 min. The membranes exhibited hemotoxicity below the limit established by ASTM F756–08. In addition, the disk diffusion tests showed antimicrobial activity against S. aureus, S. epidermidis, and P. aeruginosa strains, indicating that these membranes are promising for use in wound dressings with anti-infective biological activity.