Use of flow cytometry to compare the antimicrobial efficacy of silver-containing wound dressings against planktonic Staphylococcus aureus and Pseudomonas aeruginosa

Abstract

Silver-impregnated wound dressings continue to be routinely used for the management of infected wounds, or wounds that are at risk of becoming infected. The ability of antimicrobials that have been incorporated into wound dressings to kill microorganisms within the dressing requires appropriate evaluation using in vitro models. In vitro models that have been exploited for this purpose have included the corrected zone of inhibition and the log reduction assay. However, these and other related culturable-based assays are purported to have poor correlation with the overall microbicidal barrier activity of an antimicrobial wound dressing. This is because culturable-based methods only retrospectively indicate bacterial cell death and do not take into account viable but nonculturable states of microorganisms. Consequently, it was the purpose of this study to show that the use of flow cytometry, in conjunction with Syto(®) 9 and propidium iodide, could be used as a method for accurately evaluating and comparing the antimicrobial barrier efficacy of a silver alginate and a silver carboxymethyl cellulose dressing on individual bacterial cells without the need for the use of culturable assays. When a comparison of antimicrobial barrier efficacy on individual planktonic Staphylococcus aureus cells in a simulated wound fluid assay was made between each dressing, enhanced antimicrobial efficacy (as showed by the percentage of dead to alive bacterial cells) of the silver alginate dressing was shown. When Pseudomonas aeruginosa was exposed to both silver-containing dressings, equivalent kill rates were showed for up to 4 days. This result was not significantly different (p<0.05). By utilizing the use of flow cytometric assays, the antimicrobial barrier efficacy of wound dressings can be accurately evaluated enabling differentiation to be achieved between individual dead and live bacteria. The flow cytometric assay is considered a significant advancement to the traditionally used culturable-based methods that are presently used for antimicrobial barrier efficacy testing on planktonic microorganisms.