Eco-friendly Synthesis of Copper Sulphate Nanoparticles using Citrus Sinensis Extract and their Antimicrobial Properties: An In-vitro Study

Abstract

Introduction: The extensive use of nanoparticles underscores the importance of developing sustainable synthesis methods. There is a growing demand for green synthesis, which prioritises clean, safe, and environmentally friendly methods devoid of high temperature, pressure, energy consumption, and toxic chemicals. Aim: To produce copper sulfate nanoparticles through the utilisation of Citrus sinensis extract and assess their antibacterial efficacy against Streptococcus mutans, Staphylococcus aureus and Enterococcus faecalis. This addresses the requirement for innovative and environmentally friendly antimicrobial agents for dental purposes. Materials and Methods: This in-vitro study was conducted at the Department of Conservative Dentistry and Endodontics at Saveetha Dental College, located in Chennai, Tamil Nadu, India, during the timeframe of April 2023 to June 2023 following the acquisition of ethical clearance from the Scientific Review Board Committee. Utilising Citrus sinensis extract, a green synthesis method was employed to synthesise copper sulfate nanoparticles. Characterisation procedures included Fourier-transform Infrared Spectroscopy (FTIR) analysis, Scanning Electron Microscopy (SEM) imaging, Energy Dispersive X-ray Analysis (EDAX) analysis, and antibacterial testing. The antibacterial effectiveness of the nanoparticles was evaluated against Streptococcus mutans, Staphylococcus aureus and Enterococcus faecalis using the agar well diffusion method. The data were subjected to statistical analysis using Statistical Package for Social Sciences (SPSS) version 23.0 to assess antibacterial effectiveness. Analysis of Variance (ANOVA) was employed to analyse the Zone of Inhibition (ZOI) regarding antibacterial efficacy. Results: The copper sulfate nanoparticles, synthesised through a green approach, displayed a spherical morphology as affirmed by SEM and exhibited characteristic peaks in FTIR spectra. EDAX analysis confirmed the elemental composition of the nanoparticles. Notably, the nanoparticles showcased the most substantial antibacterial activity against Enterococcus faecalis followed by Staphylococcus aureus, and the least activity was seen against Streptococcus mutans. Conclusion: Copper sulfate nanoparticles synthesised through Citrus sinensis mediation show promising antibacterial efficacy against Enterococcus faecalis, highlighting their potential for dental applications. However, further investigations are necessary to assess their cytotoxicity, evaluate their clinical viability, and explore broader applications in antimicrobial contexts.