The main objective of this work was to prepare a tailor-made electrospun nanofibrous samples based on poly(-caprolactone) (PCL) containing tetracycline hydrochloride (TC-HCl) as a middle layer and poly(vinyl alcohol) (PVA) including phenytoin sodium (PHT-Na) as lateral layers. The characterizations of the three-layered electrospun samples were carried out by using SEM, ATR-FTIR spectroscopy along with swelling/weight loss, UV-vis spectrophotometry as well as HPLC, antibacterial and MTT tests. The SEM micrograph images showed that the average diameter of PCL nanofibers was decreased from 243 +/- 7 nm to 181 +/- 5 nm by adding TC-HCl. The hydrolytic degradation of PVA nanofibers in the exposure of phosphate buffer solution (PBS) was confirmed by ATR-FTIR results in which a change at the intensity of the characteristic peak located at 3333 cm(-1) corresponding to hydroxyl groups (OH) was observed. The UV-vis outcomes revealed a sustained control release of TC-HCl from the three-layered nanofibrous samples (PVA/PCL/PVA) with an amount of about 43% compared to the PCL nanofibers which had an ultimate release of the drug about 79%. Furthermore, the HPLC chromatograms showed the released PHT-Na from PVA nanofibers about 87%. Finally, the MTT assay along with the antibacterial evaluation exhibited that the surfaces of these electrospun three-layered nanofibrous samples have no cytotoxicity as well as the controlled release of TC-HCl from them enabled their prolonged use for preventing the bacterium growth such as S. aureus during 24-h treatment time. (c) 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43309.