Anti-Virulence Properties of Curcumin/CuO-NPs and Their Role in Accelerating Wound Healing In Vivo

<i>Background and Objectives</i>: This study introduces an innovative approach to accelerating wound healing by leveraging the bactericidal properties of mycosynthesized copper oxide nanoparticles (CuO-NPs) and their combination with curcumin against <i>Pseudomonas aeruginosa</i>. The study aims to evaluate their antimicrobial efficacy, impact on quorum sensing-associated virulence factors, and potential therapeutic applications in wound healing. <i>Materials and Methods</i>: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CuO-NPs were determined to be 25 μg/mL and 50 μg/mL, respectively. At sub-inhibitory concentrations (0.5 MIC, 0.25 MIC, and 0.125 MIC), their effects on <i>P. aeruginosa</i> growth and quorum sensing-associated virulence factors were assessed. Antioxidant activity and cytotoxicity were also evaluated. Additionally, the combination of CuO-NPs and curcumin (CUR) was tested for its enhanced wound-healing efficacy. <i>Results</i>: While CuO-NPs did not inhibit <i>P. aeruginosa</i> growth at sub-inhibitory concentrations, they significantly reduced quorum sensing-associated virulence factors in a dose-dependent manner: LasB elastase (81.8%, 60.6%, and 53.03%), LasA protease (70%, 68.5%, and 57.1%), and pyocyanin (85.7%, 71.4%, and 55.9%). CuO-NPs exhibited strong antioxidant activity by scavenging free radicals. The combination of CuO-NPs and CUR demonstrated the highest wound-healing efficacy, outperforming the negative control and Mebo ointment by 193.9% and 61.6%, respectively. Additionally, CuO-NPs exhibited selective cytotoxicity against HepG2 cancer cells while displaying minimal toxicity toward normal human skin cells. <i>Conclusions</i>: CuO-NPs, particularly in combination with CUR, show promising potential as a therapeutic agent for wound healing by inhibiting quorum sensing-associated virulence factors, exhibiting strong antioxidant activity, and demonstrating selective cytotoxicity. These findings highlight their potential biomedical applications.