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Fundamental research at the nano- and microscale to understand how biofilms interact with wounds in the presence of hyperosmotic agents

EMSL Project ID


The bactericidal effect of extracellular hyperosmosis has been acknowledged for decades1,2. Nevertheless, it was only recently shown that hyperosmosis from high NaCl concentrations effectively reduces Acinetobacter baumannii3, Enterococcus faecalis and Pseudomonas aeruginosa4 biomass and viability in vitro. In a wound, however, high NaCl concentrations can be problematic and may impair healing5. High-osmolality solutions other than NaCl (e.g., honey, glucose and other sugar pastes) are known to inhibit microbial growth, and they can be topically applied to wounds without impairing healing6. It is unclear, however, whether wound biofilms are vulnerable to the effects of hyperosmotic compounds. This knowledge is critical to developing therapeutic strategies against multidrug-resistant bacterial biofilms in wounds. The goal of this proposal is to investigate the fundamental mechanisms of wound-biofilm interactions in the presence of selected hyperosmotic agents using innovative nano- and microscale tools that have not been applied to wound biofilms before. We are a group of researchers who have significant research experience in microscale biofilm processes, nanoscale bacterial-surface interactions and mechanics, antibiotic resistance and open wound treatment. We propose to use novel and innovative technologies to advance wound biofilm research. We plan to use nuclear magnetic resonance (NMR) spectroscopy, atomic force microscopy (AFM) and microelectrodes to investigate how antibiotics penetrate into wound biofilms and how biofilms and their extracellular polymeric substances (EPS) interact with hyperosmotic agents. We propose to use confocal microscopy, cytokine assays, and Transwell migration assays to investigate how bacterial and white blood cells interact with a wound bed and discover the factors that dominate the initial cell attachment to the wound, followed by biofilm formation. We will also employ porcine tissue explants and in vivo porcine models for this work. This proposal focuses on the use of NMR and will test two of the following hypothesis for our currently funded project using NMR.

Project Details

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Principal Investigator

Haluk Beyenal
Washington State University


Ryan Renslow
Pacific Northwest National Laboratory