The potential of biogenic nano copper-loaded lipophilic quaternized chitosan in pharmacological applications: Antimicrobial, anti-inflammatory, and wound healing

Wound healing plays an important role in restoring skin function in the presence of microbial invasion. Prompt treatment is necessary to accelerate the resolution and prevent redness, exudation, or tissue necrosis. Chitosan is a biodegradable natural polymer that has a good hemostatic effect, wound repair, and antibacterial activity and has generated interest in its applications. In this study, we investigated the cytotoxicity, wound-healing potential, and molecular effects of biogenic nano-copper-loaded lipophilic quaternized chitosan nanoparticles on normal human dermal fibroblast (NHDF) cells, and evaluated their impact on cell proliferation, oxidative stress response, and gene expression. Cytotoxicity was assessed using the MTT assay and cell migration was analyzed using an in vitro scratch assay. Immunofluorescence was used to measure the levels of VEGF, PDGF, FGFR, and FN-C in NHDF and oxidative stress biomarkers, including TAC, SOD, GPx, and catalase. The gene expression levels of proinflammatory (IL-6 and IL-1 beta) and regenerative (TGF-beta) markers were quantified using RT-qPCR. Biogenic nano copper-loaded lipophilic quaternized chitosan nanoparticles enhanced NHDF cell proliferation, migration, angiogenesis, tissue repair, and extracellular matrix formation, with an EC50 value of 2.39 mu g/mL. Oxidative stress markers showed a significant increase in antioxidant enzyme activity, suggesting their protective role against cellular damage. Gene expression analysis revealed marked downregulation of inflammatory markers (IL6 and IL-1 beta) and a decrease in TGF-beta, indicating a controlled inflammatory response. Biogenic composite nanoparticles exhibit promising wound healing potential by enhancing fibroblast activity, modulating oxidative stress, and regulating key inflammatory genes. These findings suggest their suitability for advanced wound-care applications.