Rutin Incorporated Chitosan Nanocomposite Preparation and Antibacterial Activity Against Wound Pathogens: Cytoplasmic and Protein Leakage Investigation

Abstract

The rutin found in photosynthetic cells and chitosan, a fungal biopolymer, form environmentally friendly nanocomposites. Rutin chitosan nanocomposite was synthesized, and its antimicrobial capability was tested through the agar well diffusion assay and time kill curve analysis with wound pathogens like E. faecalis, E. coli, Pseudomonas sp., and S. aureus. Cytoplasmic and protein leakage analyses were performed using the cytoplasmic assay and Bradford assay. Amoxyrite was used as a standard antibiotic for comparison. In results agar well diffusion method showed that the rutin chitosan nanocomposite had a greater zone of inhibition than the control in E. faecalis and E. coli and was more effective at higher concentrations against Pseudomonas sp. and S. aureus. Time-kill curve analysis confirmed a decrease in living wound pathogen cells over time. Protein leakage analysis by the Bradford test showed higher optical density than the control at 20 µg/ml, 50 µg/ml, and 100 µg/ml but lower than the standard, with an increase in optical density at higher concentrations. Cytoplasmic leakage analysis indicated the release of cytoplasmic components from bacterial cells. Although the nanocomposite showed lower antimicrobial activity than the standard antibiotic, it significantly promoted cytoplasmic and protein leakage, highlighting its potential therapeutic applications. Rutin-chitosan nanocomposite demonstrated considerable antibacterial efficacy against P. aeruginosa, E. coli, E. faecalis, and S. aureus, confirmed by agar-well diffusion, time-kill curve, cytoplasmic leakage, and protein leakage assays. While its effects varied by concentration, it showed potential as an environmentally safe alternative to conventional antibiotics for therapeutic applications