Invitro antioxidant, Anticancer and wound healing activity of biogenically synthesised Azadirachta indica ethanolic extract copper nanoparticles

Abstract

This study explored the green synthesis of copper nanoparticles (CuNPs) using an ethanolic extract of Azadirachta indica (A. indica) and investigated their potential applications. Characterization techniques, including UV-visible spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), energy-dispersive X-ray spectroscopy (EDX), and electron microscopy, confirmed the successful synthesis of A. indica ethanolic extract of CuNPs. The antioxidant activity of the ethanolic extract of A. indica was evaluated using the stable DPPH radical method. A. indica ethanolic extract-CuNP cytotoxicity assessment of MCF-7 cells via the MTT assay revealed concentration-dependent inhibition. Apoptosis induction was evaluated using the acridine orange/ethidium bromide staining technique. In vitro wound healing activity was assessed using a scratch assay with fibroblast cells (L6 cell lines). Immunomodulatory activity was investigated by assessing the nitroblue tetrazolium (NBT)-positive cells. The nanoparticles exhibited a distinctive surface plasmon resonance peak at 679 nm and a crystalline structure with a characteristic peak at 2θ = 26.63°. A. indica ethanolic extract of CuNPs showed a dose-dependent free radical scavenging effect with an inhibition percentage ranging from 14.32% to 83.81%, and its activity was comparable to that of ascorbic acid. Cytotoxicity assessment of MCF-7 cells via MTT assay revealed concentration-dependent inhibition, with A. indica ethanolic extract of CuNPs exhibiting significant cytotoxic effects at concentrations ranging from 25 µg to 0.0976 µg. The AO/EB staining method distinguishes between living and apoptotic cells, providing insights into the various stages of apoptosis induced by the nanoparticles. A. indica ethanolic extract of CuNPs promoted the migration of fibroblasts, indicating potential wound healing properties. Immunomodulatory activity was investigated by assessing nitroblue tetrazolium (NBT)-positive cells, revealing a dose-dependent increase in immune response with A. indica ethanolic extract-CuNP concentrations ranging from 50 µg to 0.390625 µg. In conclusion, this study demonstrated the successful green synthesis of A. indica ethanolic extract-CuNPs with promising antioxidant, cytotoxic, apoptotic, wound healing, and immunomodulatory activities, suggesting their potential applications in various fields, including medicine and nanotechnology.