Povidone-Iodine and Hydrogen Peroxide Combination Improves the Anti-Biofilm Activity of the Individual Agents on Staphylococcus aureus

<i>Staphylococcus aureus</i>, particularly methicillin-resistant <i>S. aureus</i> (MRSA), poses significant challenges in healthcare settings due to its ability to form biofilms on various surfaces. These biofilms enhance bacterial survival and increase resistance to conventional treatments, complicating infection control efforts. This study evaluated the efficacy of combined povidone-iodine (PVP-I) and hydrogen peroxide (H₂O₂) to disrupt pre-formed <i>S. aureus</i> biofilms. A series of assays-including crystal violet staining, colony-forming unit (CFU) enumeration, gene expression analysis, and confocal laser scanning microscopy-were performed to assess the effects of each treatment individually and in combination. The combined treatment resulted in significantly greater reductions in biofilm biomass and viable bacteria compared with either agent alone. Gene expression analysis revealed downregulation of key biofilm-associated genes (<i>icaA</i>, <i>icaB</i>, <i>icaD</i>, <i>icaR</i>, and <i>clfA</i>), suggesting interference with biofilm stability and maintenance. While formal synergy quantification was not conducted, the observed effects suggest a potentially synergistic or additive interaction between the two agents. These findings support the use of dual antiseptic strategies as a promising approach to biofilm eradication and highlight the potential clinical utility of dual antiseptic strategies. However, we underscore the need for further optimization and safety evaluation.