3D Nanofibrous Scaffolds for Encapsulation-Controlled Vancomycin Delivery: Antibacterial Performance and Cytocompatibility

This study aimed to engineer nanofibrous scaffolds that prioritize architecture, rather than relying solely on the drug, to achieve reproducible, long-acting local therapies. Cotton-wool-like fiber, three-dimensional (3D) poly(L-lactic acid)/polyethene glycol (PLLA/PEG) blend scaffolds were fabricated using solution blow spinning (SBS) as a customizable encapsulation platform for controlled antibiotic release. Morphological and wettability analyses were performed by scanning electron microscopy (SEM) and pendant-drop contact angle measurements, respectively. Fiber diameters were quantified using ImageJ. The chemical composition and thermal behavior were investigated by Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). In vitro, assays were conducted to assess the antimicrobial activity of vancomycin-loaded scaffolds against <i>Staphylococcus aureus</i> (disk diffusion method), as well as their cytocompatibility (Live/Dead assay in Vero cells) and hemocompatibility (ASTM F756-17 hemolysis test). All biological data were statistically analyzed using ANOVA with Tukey's post-test, Mann-Whitney, and paired <i>t</i>-tests, with significance set at <i>p</i> ≤ 0.05. Structural optimization identified PLLA/PEG 85:15 as the most stable composition, producing homogeneous mats with high porosity and rapid wettability. Incorporation of vancomycin (10 wt.%) reduced the fiber diameter (0.23 ± 0.11 µm) compared with unloaded scaffolds (0.32 ± 0.17 µm), indicating drug-polymer interactions that modulated jet elongation. FTIR, DSC, and TGA analyses confirmed polymer miscibility and stabilization of VMC within the fibrous matrix, with no signs of degradation. Drug release exhibited a biphasic profile, with an initial burst during the first 72 h. PLLA/PEG-VMC scaffolds produced larger inhibition zones against <i>S. aureus</i> (18.55 mm ± 1.2 to 6.63 mm ± 0.2 at 120 h) compared with free VMC (12.91 mm ± 3.8 to 4.07 mm ± 0.6291), while blank scaffolds were inactive. Hemolysis remained within the range 2% < PLLA/PEG-VMC < 5%, indicating acceptable hemocompatibility according to ASTM standards. Although VCM-loaded PLLA/PEG scaffolds slightly reduced Vero cell viability, no statistically significant differences were observed compared with the control group. These findings demonstrate that the architecture of nanofibers presents itself as a potential platform for antimicrobial therapy with topical vancomycin in potential applications such as wound dressings or implant coatings.