Sequential Hypertonic-Hypotonic Treatment Enhances Efficacy of Antibiotic against Acinetobacter baumannii Biofilm Communities

Infections with bacterial biofilm communities are highly tolerant of antibiotics. This protection is attributed, in part, to a hydrated extracellular polymeric substance (EPS) that surrounds the bacterial community and that limits antibiotic diffusion. In this study, we evaluated whether it is possible to dehydrate and then re-hydrate a biofilm as a means to increase antibiotic penetration and efficacy. Acinetobacter baumannii biofilms (24 h) were exposed to hypertonic concentrations of maltodextrin, sucrose or polyethylene glycol (PEG) as the dehydration step. These biofilms were then washed with deionized water containing 10 times the concentration of antibiotics needed to kill these bacteria in broth culture (50 µg/mL tobramycin, 300 µg/mL chloramphenicol, 20 µg/mL ciprofloxacin or 100 µg/mL erythromycin) as the rehydration step. Biofilms were then harvested, and the number of viable cells was determined. Sequential treatment with PEG and tobramycin reduced cell counts 4 to 7 log (p < 0.05) relative to combining PEG and tobramycin in a single treatment, and 3 to 7 log relative to tobramycin treatment alone (p < 0.05). Results were variable for other osmotic compounds and antibiotics depending on the concentrations used, likely related to mass and hydrophobicity. Our findings support future clinical evaluation of sequential regimens of hypertonic and hypotonic solutions to enhance antibiotic efficacy against chronic biofilm infections.