R Cooper1, L Jenkins2, S Hooper1. 1. PhD, Cardiff School of Health Sciences, Cardiff Metropolitan University, Cardiff, Wales, UK. 2. BSc, Cardiff School of Health Sciences, Cardiff Metropolitan University, Cardiff, Wales, UK.
Abstract
OBJECTIVE: Pseudomonas aeruginosa has been linked to chronic wound infections, where its ability to form biofilms and to tolerate antimicrobial agents helps to facilitate its persistence. This study aimed to investigate the susceptibility of biofilms of Pseudomonas aeruginosa to Medihoney in vitro. METHOD: Biofilms were cultivated in microtitre plates with and without a range of concentrations of Medihoney, and effects on biofilm were monitored by optical density (at 650nm), biomass (by staining with crystal violet), metabolic activity (using an esterase assay) and viability (by determining total cell counts). Structural effects on established biofilms were examined by scanning electron microscopy and epifluorescence following staining by LIVE/DEAD® BacLight, which also showed effects on vitality. RESULTS: The lowest concentration of Medihoney found to prevent biofilm formation was 17%(w/v), whereas on average 35.5%(w/v) of Medihoney was required to inhibit established biofilms. Susceptibility did not vary with length of biofilm establishment between 24 and 72 hours. Extensive structural changes in established biofilms were seen in the sample with less than or equal to 30%(w/v) Medihoney using scanning electron microscopy and loss of viability was found in test samples with less than or equal to 20%(w/v) Medihoney concentration using fluorescent staining, together with loss of biofilm structure. CONCLUSION: Using a range of methods to evaluate biofilm integrity, this study demonstrates that Medihoney inhibits Pseudomonas aeruginosa biofilms in vitro at concentrations that are attainable in clinical use. Whether Medihoney has the potential to disrupt Pseudomonas aeruginosa biofilms in cutaneous wounds must now be tested in patients. DECLARATION OF INTEREST: This study was sponsored by Derma Sciences Inc, NJ. An unrestricted grant was provided and the sponsors were not involved in the design of the experiments or the preparation of this manuscript.
OBJECTIVE:Pseudomonas aeruginosa has been linked to chronic wound infections, where its ability to form biofilms and to tolerate antimicrobial agents helps to facilitate its persistence. This study aimed to investigate the susceptibility of biofilms of Pseudomonas aeruginosa to Medihoney in vitro. METHOD: Biofilms were cultivated in microtitre plates with and without a range of concentrations of Medihoney, and effects on biofilm were monitored by optical density (at 650nm), biomass (by staining with crystal violet), metabolic activity (using an esterase assay) and viability (by determining total cell counts). Structural effects on established biofilms were examined by scanning electron microscopy and epifluorescence following staining by LIVE/DEAD® BacLight, which also showed effects on vitality. RESULTS: The lowest concentration of Medihoney found to prevent biofilm formation was 17%(w/v), whereas on average 35.5%(w/v) of Medihoney was required to inhibit established biofilms. Susceptibility did not vary with length of biofilm establishment between 24 and 72 hours. Extensive structural changes in established biofilms were seen in the sample with less than or equal to 30%(w/v) Medihoney using scanning electron microscopy and loss of viability was found in test samples with less than or equal to 20%(w/v) Medihoney concentration using fluorescent staining, together with loss of biofilm structure. CONCLUSION: Using a range of methods to evaluate biofilm integrity, this study demonstrates that Medihoney inhibits Pseudomonas aeruginosa biofilms in vitro at concentrations that are attainable in clinical use. Whether Medihoney has the potential to disrupt Pseudomonas aeruginosa biofilms in cutaneous wounds must now be tested in patients. DECLARATION OF INTEREST: This study was sponsored by Derma Sciences Inc, NJ. An unrestricted grant was provided and the sponsors were not involved in the design of the experiments or the preparation of this manuscript.
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