| Summary: | In this study, we aimed to explore the morphologies of porous poly(lactic acid) (PLA) fibers through liquid–liquid phase separation, and investigate the relationship among pore formation, physical properties, and antibacterial activities of the fibers for identifying their potential as drug delivery carriers. Antibacterial activities of gentamicin-, kanamycin-, and amikacin-loaded PLA fibers against <i>E. coli</i> and <i>S. epidermidis</i> were evaluated. The antibacterial activity of drugs against <i>E. coli</i> showed the following profile: gentamicin > amikacin > kanamycin; however, <i>S. epidermidis</i> growth was almost completely inhibited immediately after the administration of all three drugs. The efficiency of gentamicin can be attributed to the electrostatic interactions between the positively and negatively charged antibiotic and bacterial cell membrane, respectively. Furthermore, gentamicin-loaded porous PLA fibers were evaluated as drug delivery systems. The cumulative amount of gentamicin in porous PLA nanofibers was considerably higher than that in other PLA fibers for 168 h, followed by 7:3 PLA > 6:4 PLA > 5:5 PLA > non-porous PLA. The 7:3 PLA fibers were projected to be ideal drug carrier candidates for controlled antibiotic release in delivery systems owing to their interconnected internal structure and the largest surface area (55.61 m<sup>2</sup> g<sup>−1</sup>), pore size (42.19 nm), and pore volume (12.78 cm<sup>3</sup> g<sup>−1</sup>).
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