<i>In Vitro</i> Antibacterial Activity of Hydrogen Peroxide and Hypochlorous Acid, Including That Generated by Electrochemical Scaffolds

Abstract

Hydrogen peroxide (H₂O₂) and hypochlorous acid (HOCl) are biocides used for cleaning and debriding chronic wound infections, which often harbor drug resistant bacteria. Here, we evaluated the <i>in vitro</i> activity of H₂O₂ and HOCl against 27 isolates of eight bacterial species involved in wound infections. Minimum inhibitory concentrations (MICs) and minimum biofilm bactericidal concentrations (MBBCs) were measured. When compared to their respective MICs, MBBCs of isolates exposed to H₂O₂ were 16- to 1,024-fold higher and those exposed to HOCl were 2- to 4-fold higher. We evaluated selection of resistance after exposure of <i>Staphylococcus aureus</i> and <i>Pseudomonas aeruginosa</i> biofilms to 10 iterations of electrochemically generated HOCl or H₂O₂ delivered using electrochemical scaffolds (e-scaffolds), observing no decrease in anti-biofilm effects with serial exposure to e-scaffold-generated H₂O₂ or HOCl. 24-hour exposure to H₂O₂-generating e-scaffolds consistently decreased colony forming units (CFUs) of <i>S. aureus</i> and <i>P. aeruginosa</i> biofilms by ∼5.0-log₁₀ and ∼4.78-log₁₀ through 10 iterations of exposure, respectively. 4-hour exposure to HOCl-generating e-scaffolds consistently decreased CFUs of <i>S. aureus</i> biofilms by ∼4.9-log₁₀, and 1-hour exposure to HOCl-generating e-scaffolds consistently decreased CFUs of <i>P. aeruginosa</i> biofilms by ∼1.57-log₁₀ These results suggest that HOCl has similar activity against planktonic and biofilm bacteria, whereas the activity of H₂O₂ is less against biofilm than planktonic bacteria, and that repeat exposure to either biocide, generated electrochemically under the experimental conditions studied, does not lessen antibiofilm effects.