Evaluating the Combined Effects of Zinc Oxide and Hamamelitannin Nanoparticles on Wound Healing: Results from In Vitro and Zebrafish Experiments

Abstract

This study evaluates the combined effects of zinc oxide (ZnO) and hamamelitannin (HMT) nanoparticles on wound healing, focusing on their antimicrobial, anti-inflammatory, antioxidant, and molecular mechanisms. Antimicrobial assays revealed significant activity of Zinc Oxide Hamamelitannin Nanoparticles (ZnO-HM NPs) against wound-associated pathogens Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Vibrio vulnificus demonstrating enhanced bacterial inhibition compared to individual components. Anti-inflammatory properties were assessed through protein denaturation inhibition using ibuprofen as a reference, with ZnO-HM NPs showing strong inhibition of protein denaturation. Antioxidant activity, evaluated by DPPH free radical scavenging, revealed high efficacy in reducing oxidative stress.  Molecular docking studies were conducted against key proteins 7S53 (antimicrobial protein), 4NG2 (oxidative stress regulator), 20MF (inflammatory mediator), and 5TV2 (wound healing protein), demonstrating strong binding affinities, suggesting potential mechanisms for their biological effects. Additionally, RT-PCR analysis on wounded zebrafish tissues showed significant downregulation of pro-inflammatory markers including mmp13, TNF-α, and IL-1β, further supporting the anti-inflammatory action of the nanoparticles. In vivo zebrafish models revealed accelerated wound closure and tissue regeneration, confirming the synergistic effects of ZnO-HM NPs on wound healing. These results suggest that ZnO-HM NPs hold significant promise as a multifunctional therapeutic agent for enhancing wound healing through antimicrobial, antioxidant, and anti-inflammatory pathways, reinforced by molecular targeting.