Green synthesis and chemical characterization of copper nanoparticles using Allium saralicum leaves and assessment of their cytotoxicity, antioxidant, antimicrobial, and cutaneous wound healing properties

Abstract

In recent decades, nanotechnology is growing rapidly owing to its widespread application in science and industry. The aim of the experiment was the green synthesis of copper nanoparticles using Allium saralicum R.M. Fritsch aqueous extract and assessment of their cytotoxicity, antioxidant, antibacterial, antifungal, and cutaneous wound healing effects under in vitro and in vivo conditions. These nanoparticles were characterized by Fourier transformed infrared spectroscopy (FT‐IR), UV–visible spectroscopy, field emission scanning electron microscopy (FE‐SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM). DPPH free radical scavenging test was done to assess the antioxidant properties, which indicated similar antioxidant potentials for CuNPs@Allium and butylated hydroxytoluene. Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and Minimum Fungicidal Concentration (MFC) were specified by macro‐broth dilution assay. CuNPs@Allium indicated higher antibacterial and antifungal effects than all standard antibiotics (p ≤ 0.01). Also, CuNPs@Allium inhibited the growth of all bacteria at 1–8 mg/ml concentrations and removed them at 2–8 mg/ml concentrations (p ≤ 0.01). In the case of antifungal properties of CuNPs@Allium, they prevented the growth of all fungi at 1–4 mg/ml concentrations and destroyed them at 2–8 mg/ml concentrations (p ≤ 0.01). In vivo experiment, after creating the cutaneous wound, the rats were randomly divided into six groups (n = 10): untreated control, treatment with Eucerin basal ointment, treatment with 3% tetracycline ointment, treatment with 0.2% CuSO4 ointment, treatment with 0.2% A. saralicum ointment, and treatment with 0.2% CuNPs@Allium ointment. Use of CuNPs@Allium ointment in the treatment groups substantially reduced (p ≤ 0.01) the wound area, total cells, neutrophil, macrophage, and lymphocyte and remarkably raised (p ≤ 0.01) the wound contracture, hydroxyl proline, hexosamine, hexuronic acid, fibrocyte, and fibrocytes/fibroblast rate compared to other groups. The synthesized CuNPs@Allium had high cell viability dose‐dependently (Investigating the effect of the plant on HUVEC cell line) and revealed this method was nontoxic. The results revealed the useful non‐cytotoxic, antioxidant, antibacterial, antifungal, and cutaneous wound healing effects of CuNPs@Allium.