Wound healing activity of green synthesized copper nanoparticles through cell proliferation-migration, antimicrobial effects, and nitric oxide triggering.

Abstract

The present experimental study aimed to assess the in vitro wound healing and anti-inflammatory effects of green synthesized copper nanoparticles (CuNPs) by the methanol extract of Ferula macrecolea (Boiss), as a plant with various pharmacological effects, such as anti-inflammatory and antimicrobial effects, in traditional and modern medicine. The precipitation approach was used for the green synthesis of CuNPs by mixing the methanol and copper sulfate solution. Cell viability and fibroblast proliferation assay were performed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. The migration abilities of fibroblast cells were evaluated using the in vitro scratch assay for wound healing. The effects of CuNPs on gene expression of inducible nitric oxide synthesis (iNOS) were also examined by real-time polymerase chain reaction (PCR). In vitro antibacterial susceptibility test of CuNPs was carried out according to the standards protocol of the National Committee for Clinical Laboratory Standards. The scanning electron microscope analysis revealed that the green synthesized CNP exhibited a globular shape with a size ranging from 15 to 90 nm, while the majority were at 40-60 nm. The results of the MTT assay demonstrated that the calculated 50% cytotoxic concentration (CC₅₀) value of green synthesized CuNPs was 236.3 μg/mL. The optimum concentrations of the CuNPs were selected based on the CC₅₀, which dose-dependently increased the proliferation of fibroblast cells. The CuNPs dose-dependently increased the rate of wound closure after 16 and 24 h. The results of the real-time PCR illustrated that CuNPs caused upregulation in the expression level of the iNOS gene in RAW 264.7 cells. CuNPs showed promising antimicrobial effects against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa. The present study highlighted the high potency of green CuNPs synthesized by F. macrecolea for wound healing through their antimicrobial properties, proliferation of fibroblast cells, and provoking iNOS.