Benzyl isothiocyanate-loaded chitosan beads: a novel strategy to combat biofilm formation by <i>Staphylococcus aureus</i> and <i>Escherichia coli</i>

Aims This study aimed to develop and evaluate chitosan beads loaded with benzyl isothiocyanate (BITC) as a novel therapeutic strategy to combat biofilm-associated infections caused by Staphylococcus aureus and Escherichia coli.Methods The beads were fabricated via ionic gelation and characterized using Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and ultraviolet-visible spectrophotometry (UV-Vis). Antibacterial and antibiofilm activities of the optimal formulation BITC-loaded beads (B-Cs2) were assessed. Supporting computational analyses (molecular docking) were carried out; detailed methods and results are provided in the Supplementary Material.Results B-Cs2 beads (10 mg) exhibited strong antibacterial activity, producing inhibition zones of 34.0 ± 1.0 mm for S. aureus and 24.0 ± 2.0 mm for E. coli. The formulation also significantly inhibited biofilm formation, reducing it by 49.3% in S. aureus and 48.2% in E. coli. Molecular docking predicted potential interactions with key virulence proteins, suggesting that BITC may disrupt distinct virulence pathways; however, further biochemical validation is required to confirm these proposed mechanisms.Conclusion BITC-loaded chitosan beads function as a dual-action system with potent antibacterial and antibiofilm properties, demonstrating significant potential for applications in clinical therapeutics and food safety.