Biogenic carbon dots: a novel mechanistic approach to combat multidrug-resistant critical pathogens on the global priority list.

This study delves into investigating alternative methodologies for anti-microbial therapy by focusing on the mechanistic assessment of carbon dots (CDs) synthesized from F. benghalensis L. extracts. These biogenic CDs have shown remarkable broad-spectrum anti-bacterial activity even against multi-drug resistant (MDR) bacterial strains, prompting a deeper examination of their potential as novel anti-microbial agents. The study highlights the significant detrimental impact of CDs on bacterial cells through oxidative damage, which disrupts the delicate balance of ROS control within the cells. Notably, even at low doses, the anti-bacterial activity of CDs against MDR strains of P. aeruginosa and E. cloacae is highly effective, demonstrating their promise as potent antimicrobial agents. The research sheds light on the capacity of CDs to generate ROS, leading to membrane lipid peroxidation, loss of membrane potential, and rupture of bacterial cell membranes, resulting in cytoplasmic leakage. SEM and TEM analysis revealed time-dependent cell surface, morphological, and ultrastructural changes such as elongation of the cells, irregular surface protrusion, cell wall and cell membrane disintegration, internalization, and aggregations of CDs. These mechanisms offer a comprehensive explanation of how CDs exert their anti-bacterial effects. We also determined the status of plasma membrane integrity and evaluated live (viable) and dead cells upon CD exposure by flow cytometry. Furthermore, comet assay, biochemical assays, and SDS PAGE identify DNA damage, carbohydrate and protein leakage, and distinct differences in protein expression, adding another layer of understanding to the mechanisms behind CDs' anti-bacterial activity. These findings pave the way for future research on managing ROS levels and developing CDs with enhanced anti-bacterial properties, presenting a breakthrough in anti-microbial therapy.