In this study, the lavender essential oil-incorporated polylactic acid (PLA) nanofibers were fabricated by the electrospinning technique with the presence of kolliphor as a nonionic surfactant. The FTIR spectra supported the chemical composition of the fibers. The FTIR spectra demonstrated that there is no chemical reaction present between PLA and lavender essential oil. The SEM images of all nanofibers showed bead-free morphology. ImageJ results showed that the average diameter of lavender oil-loaded fibers ranged between 121.6 ± 32 and 228.2 ± 53 nm. All lavender essential oil-incorporated nanofibers were hydrophobic with satisfactory thermal properties. Furthermore, the lavender essential oil-incorporated PLA nanofibrous mats exhibited good antioxidant activity. The results showed that as the concentration of the essential oil in resulting nanofibers increased, the antioxidant activity also increased. According to the results of this study, lavender essential oil-loaded PLA fibers can be considered for a wide range of potential applications such as active packaging, food coating, facial masks, and wound dressing.