Presentation Title

Identifying Biofilm-Disrupting Properties of N-acetyl-L-cysteine

Start Date

November 2016

End Date

November 2016

Location

HUB 302-#180

Type of Presentation

Poster

Abstract

Elderly and diabetic patients have increased risk of developing chronic wounds because they have reduced antioxidant activity, increased oxidative stress, and biofilm-producing bacteria in the wound. We generated chronic wounds in diabetic mice by inhibiting two major enzymes that decrease oxidative stress. We found that the excess oxidative stress can be reversed through treatment with N-acetyl-L-cysteine (NAC), a commonly used chemical to loosen mucus in cystic fibrosis patients. We hypothesized that NAC disrupts biofilm by chemically interacting with different components of the extracellular polymeric substance secreted by biofilm-producing bacteria. To test this possibility, we treated mature biofilm from diabetic mouse chronic wounds in vitro in various ways and analyzed the effects on protein. To test whether the thiol group in NAC is critical to disrupt biofilm, we compared the ability of NAC to other thiol-containing chemicals, dithiothreitol and β-mercaptoethanol, and found that only NAC produced a smaller molecular weight protein band pattern, suggesting that the thiol group is not important. To determine whether the carboxyl group is key for NAC biofilm disruption, we used N-acetylcysteine amide (AD4), a NAC derivative with an amide group replacing the acidic carboxyl of NAC. We found that AD4 produced similar band patterns to the control; hence, the NAC carboxyl group appears to be critical for biofilm dismantling. We tested NAC at neutral pH and found that it produced similar protein bands to control. These results highlight the importance of pH-balance in the biofilm and that the carboxyl group of NAC may be critical for biofilm disruption. Research on biofilms in chronic wounds is needed because it is a major problem in these wounds. Our chronic wound model allows investigation of early-occurring processes that are crucial for biofilm growth and development of methods to dismantle biofilm, which would significantly advance the treatment of chronic wounds.

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Identifying Biofilm-Disrupting Properties of N-acetyl-L-cysteine

HUB 302-#180

Elderly and diabetic patients have increased risk of developing chronic wounds because they have reduced antioxidant activity, increased oxidative stress, and biofilm-producing bacteria in the wound. We generated chronic wounds in diabetic mice by inhibiting two major enzymes that decrease oxidative stress. We found that the excess oxidative stress can be reversed through treatment with N-acetyl-L-cysteine (NAC), a commonly used chemical to loosen mucus in cystic fibrosis patients. We hypothesized that NAC disrupts biofilm by chemically interacting with different components of the extracellular polymeric substance secreted by biofilm-producing bacteria. To test this possibility, we treated mature biofilm from diabetic mouse chronic wounds in vitro in various ways and analyzed the effects on protein. To test whether the thiol group in NAC is critical to disrupt biofilm, we compared the ability of NAC to other thiol-containing chemicals, dithiothreitol and β-mercaptoethanol, and found that only NAC produced a smaller molecular weight protein band pattern, suggesting that the thiol group is not important. To determine whether the carboxyl group is key for NAC biofilm disruption, we used N-acetylcysteine amide (AD4), a NAC derivative with an amide group replacing the acidic carboxyl of NAC. We found that AD4 produced similar band patterns to the control; hence, the NAC carboxyl group appears to be critical for biofilm dismantling. We tested NAC at neutral pH and found that it produced similar protein bands to control. These results highlight the importance of pH-balance in the biofilm and that the carboxyl group of NAC may be critical for biofilm disruption. Research on biofilms in chronic wounds is needed because it is a major problem in these wounds. Our chronic wound model allows investigation of early-occurring processes that are crucial for biofilm growth and development of methods to dismantle biofilm, which would significantly advance the treatment of chronic wounds.