Biogenic Synthesis of Chitosan/ CuO Nanocomposite From Azadirachta indica Extract: A Multifunctional Agent for Biomedical Applications

ABSTRACT This study reports the green synthesis of a chitosan/copper oxide (CS/CuO) nanocomposite using Azadirachta indica leaf extract. HRTEM imaging revealed well‐dispersed spherical nanoparticles with an average diameter of 44.57 nm, while XRD analysis confirmed the presence of a cubic crystalline structure. FESEM analysis revealed a uniform distribution of CuO nanoparticles on the chitosan surface, indicating enhanced interfacial properties and structural integrity. The CS/CuO nanocomposite exhibits potent antibacterial activity, producing a maximum inhibition zone of 35.10 mm against Pseudomonas aeruginosa at 100 μg/mL. The nanocomposite demonstrated strong antioxidant activity, achieving 86.81% DPPH radical scavenging at 1000 μg/mL and an IC 50 of 261.64 μg/mL. Cell viability assays using various concentrations of CS/CuO nanocomposites on GM00637 cells revealed cytotoxicity and enhanced colony formation. mRNA expression analysis indicated significant changes in target gene expression in GM00637 cells. The study evaluated the wound‐healing effectiveness of a chitosan/copper oxide (CS/CuO) nanocomposite using an excision wound model in mice. Results showed a significantly faster healing rate in treated groups compared to the untreated control group. After 10 days of treatment, 74.15% of the wounds healed, with complete healing observed within 15 days for the mice receiving the CS/CuO nanocomposite.