Gold nanoparticle conjugation enhances berberine's antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA)

Nanoparticle (NP) conjugation with various biomolecules is one of the most promising approaches for targeting Methicillin-resistant Staphylococcus aureus (MRSA). In this study, berberine (BER) was conjugated with gold nanoparticles (AuNPs) to enhance its antibacterial activity against MRSA. Chemically synthesized AuNPs were characterized by UV–vis spectroscopy, size distribution and Field Emission-Scanning Electron Microscope (FE-SEM) analysis. Berberine was conjugated with AuNPs and the conjugants were characterized using UV–vis spectroscopy and Fourier Transform Infrared (FTIR). The cytotoxicity of free and conjugated BER was also investigated. Comparative studies were conducted based on the Minimum Inhibitory Concentration (MIC) and anti-biofilm activities of conjugants and free BER against MRSA isolates. To verify cell membrane disruption and intracellular imbalance following treatment exposure, reactive oxygen species (ROS) and live-dead staining experiments were performed. In vivo antibacterial efficacy of treated groups was also assessed in a BALB/c mouse-infected skin model. DLS measurement, FE-SEM, and UV–vis spectroscopy confirmed the synthesis of AuNPs with a narrow size distribution of 49.38 nm and a zeta potential of −31.9 mV. The results from UV–vis spectroscopy and FTIR provided support for the functionalization of AuNPs by BER functional groups. The In vitro antibacterial results demonstrated that the conjugated BER exhibited a lower MIC value against MRSA (109.5 μg/ml) compared to free BER (165 μg/ml). Free and conjugated BER, at their MIC concentrations, demonstrated anti-biofilm activity, resulting in biofilm eradication of 13.9 and 22.33 %, respectively. The highest level of ROS production (93 %) was associated with the conjugated BER at a concentration of 27.37 μg/ml. This finding indicates a disruption in cell membrane integrity and a reduction in bacterial viability, as demonstrated by ROS and live/dead staining assays. The cytotoxicity study on the mouse L929 fibroblast cell line revealed approximately 100 % cell viability when exposed to free or conjugated BER at their MIC concentration. This result indicates the biosafety of both of the compounds. The in vivo study in the infected skin model groups treated with conjugated and free BER revealed MRSA survival rate of 2.7 % and 26 %, respectively. These findings suggest that conjugated BER could be an effective nanoformulation candidate with a potential role in managing MRSA associated infections.