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Staphylococcus aureus is the leading cause of skin infections in the U.S., and its rapid evolution and resistance to antibiotics create a barrier to effective treatment. In this study, we engineered a composite membrane with bacterial cellulose and carbon nanotubes (BC-CNT) as an electroactive dressing to rapidly eradicate vancomycin-intermediate S. aureus. Nonpathogenic Komagataeibacter sucrofermentans produced the BC membrane at an air-liquid interface. Then, carboxyl-functionalized multi-walled CNTs were integrated into decellularized BC to create stable and electrically conductive BC-CNT dressings. The electric potential and ionic flux across BC-CNT were modeled and standardized via chronoamperometry for experimental validation. We found that treatment with electroactive BC-CNT increases S. aureus sensitivity to vancomycin and prevents macro-scale biofilm formation. The bactericidal efficacy of the composite membrane is consistent with electrochemical stress caused by voltage mediated with BC-CNT. After a single hour of combinatorial electrical and drug treatment, biofilm-forming capacity was inhibited by nearly 92 %. These results advance applications of electrochemistry in medicine and create a new direction to overcome S. aureus infections on skin and soft tissues. 

IntroductionChronic lung infection due to bacterial biofilms is one of the leading causes of mortality in cystic fibrosis (CF) patients. Among many species colonizing the lung airways,Pseudomonas aeruginosaandStaphylococcus aureusare two virulent pathogens involved in mechanically robust biofilms that are difficult to eradicate using airway clearance techniques like lung lavage. To remove such biological materials, glycoside hydrolase-based compounds are commonly employed for targeting and breaking down the biofilm matrix, and subsequently increasing cell susceptibility to antibiotics. Materials and methodsIn this study, we evaluate the effects of N-acetyl cysteine (NAC) and Cysteamine (CYST) in disrupting interfacial bacterial films, targeting different components of the extracellular polymeric substances (EPS). We characterize the mechanics and structural integrity of the interfacial bacterial films using pendant drop elastometry and scanning electron microscopy. Results and discussionOur results show that the film architectures are compromised by treatment with disrupting agents for 6 h, which reduces film elasticity significantly. These effects are profound in the wild type and mucoidP. aeruginosa, compared toS. aureus. We further assess the effects of competition and cooperation betweenS. aureusandP. aeruginosaon the mechanics of composite interfacial films. Films ofS. aureusand wild-typeP. aeruginosacocultures lose mechanical strength while those ofS. aureusand mucoidP. aeruginosaexhibit improved storage modulus. Treatment with NAC and CYST reduces the elastic property of both composite films, owing to the drugs’ ability to disintegrate their EPS matrix. Overall, our results provide new insights into methods for assessing the efficacy of mucolytic agents against interfacial biofilms relevant to cystic fibrosis infection.