Hebah Ayash, Gülşah Esen, Ece Sabuncu, İnci Deniz, Pınar Akkuş Süt, Gökçen Yaşayan, Hande Sipahi, Ahmet Aydın, Soodabeh Davaran, Muhammed Hamitoğlu
Advanced wound dressings should actively promote tissue repair while preventing microbial infection and excessive inflammation. In this study, multifunctional electrospun nanofibers composed of poly(vinyl alcohol)/chitosan/gelatin were developed by incorporating green-synthesized mesoporous silica nanoparticles (MSNs), Acacia gum, and Hypericum perforatum extract (HPE). Biological activity and safety were evaluated using disk-diffusion antimicrobial testing, MTT cell viability assays, nitric oxide measurement in LPS-stimulated RAW 264.7 macrophages, an L929 fibroblast scratch assay, and Ames and comet assays. The optimized nanofibers exhibited a uniform morphology with an average fiber diameter of approximately 400 nm and high encapsulation efficiencies (86-96%). MSN incorporation significantly prolonged hypericin release, producing a biphasic release profile with approximately 1.4-fold slower release than nanofibers without MSNs, while increasing Young's modulus from 2.1 to 2.5 MPa. The hybrid nanofibers demonstrated broad-spectrum antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, and Candida albicans, achieving approximately 70% of the efficacy of standard antimicrobial agents. In lipopolysaccharide-stimulated RAW 264.7 macrophages, the optimized formulation reduced nitric oxide production by approximately 51%, comparable to indomethacin and L-NAME. Although fibroblast migration was not significantly enhanced, the nanofibers exhibited acceptable cytocompatibility and showed no mutagenic or genotoxic effects in Ames and comet assays. These findings demonstrate that green-synthesized MSN-enhanced herbal nanofibers represent a safe and promising platform for sustained drug delivery and advanced wound-healing applications.