Nanofiber membranes based on biocompatible polymers offer advantages such as high surface area and porosity that allow them to support active ingredients, such as drugs with antibiotic activity, such as neomycin, which have potential as a treatment for several bacteria. The technique used to produce these membranes can be electrospinning, which in turn has two modifications to optimize the release of the drug, such as simple and coaxial needles: the simple technique produces 145 ± 22 nm fibers. In contrast, in the coaxial technique at 119 ± 36 nm, the infrared spectra revealed electrostatic interactions that allowed the release of neomycin, which enabled delivery. In turn, the release tests of the coaxial system revealed that in the first hour, approximately 40% of the bacteria were released via a degradation release mode according to Kerseymeres-Peppas Kinetic, which is in accordance with the antibacterial activity, which was 100% effective at that time for gram-positive and gram-negative bacteria.