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

Abstract

The present experimental study was designed to evaluate the in vitro wound healing and anti-inflammatory effects of copper nanoparticles (CuNPs) green synthesized by the methanol extract 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 to green synthesis of CuNPs through 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 fibroblasts cells were evaluated using a cell scratch in vitro wound healing assay. Effects of CuNPs on iNOS expression genes were also performed by Real-time PCR. In vitro antibacterial susceptibility test of CuNPs was carried out according to the standards protocol of the National Committee for Clinical Laboratory Standards (NCCLS). The analysis of SEM revealed that the green synthesized CNP exhibited a globular shape with a size ranging from 15-90 nm; while their majority were at 40-60 nm. The results of the MTT assay showed that the calculated CC50 value of green synthesized CuNPs was 236.3 μg/mL. The optimum concentrations of the CuNPs which were selected based on the 50% cytotoxic concentration (CC50) value; whereas 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 showed that CuNPs causes an upregulation in expression level of gene of nitric oxide synthesis (iNOS) in RAW 264.7 cells. CuNPs showed the promising antimicrobial effects against S. aureus, S. epidermidis and Ps. aeruginosa. The present study showed the high potency of CuNPs green synthesized by F. macrecolea for wound healing through their antimicrobial, proliferation of fibroblast cells, and provoking iNOS.