Résumé : Background/Objectives: Polymeric barrier membranes (BMs) are usually used in guided bone regeneration to isolate the bone defect from the surrounding tissue, favoring bone apposition. This study proposes a third-generation BM made of polycaprolactone (PCL), loaded with different concentrations of nano-hidroxyapatite (nHAP) and gentamicin (GEN), and fabricated by electrospinning. Methods: The mechanical properties of the polymer, together with the fabrication procedure, offer porosity with interconnectivity to permit cell adhesion and proliferation. Bacterial contamination of the BM can induce infection at the bone level, leading to unfavorable clinical outcomes of the regeneration procedure. Results: Therefore, BMs have been proposed as carriers for local GEN antibiotic therapy, demonstrating antibacterial properties against S. aureus, S. mutans, and P. aeruginosa, depending on the drug concentration, while being negligibly affected by the nHAP content. X-ray diffraction, FTIR-ATR, and SEM allowed for BM structural characterization, demonstrating the presence of GEN/nHAP and establishing the fiber diameter, which influences the mechanical properties in dry and wet conditions and the drug release behaviorA BM cytotoxicity assessment, performed over 1 and 5 days, revealed that a high nHAP concentration provided protection against cytotoxicity, in contrast to GEN, and that cell proliferation and cell adhesion increased in the presence of nHAP. The BM’s bioactivity was demonstrated by mineralization after 21 days in simulated body fluid in an SEM/EDX analysis. Conclusions: The electrospun 15 wt.% nHAP and 2 wt.% GEN-loaded third-generation BM could be a promising alternative for guided bone regeneration.