Monitoring Of Biofilm Formation In Burn Wounds In Real-time
Angela Gibson, Anna Garren, Stephanie Metzger, Lindsay Kalan.
University of Wisconsin, Madison, WI, USA.
Background: Burn wound healing can be enhanced by daily mechanical debridement and topical antimicrobials to remove potential slough and prevent infection. However, if biofilms form they are recalcitrant to antimicrobials and can impact wound healing by impairing granulation tissue formation and re-epithelialization. The incidence of biofilm formation in human burn wounds is understudied and predicting burn wounds more susceptible to biofilm is challenging. Here we describe the development of a biofilm model of human burn wounds to study infection dynamics and response to treatments.
Methods:Human skin discarded after elective surgical procedures was burned ex-vivo using a metal soldering device and placed into tissue culture medium. After 24 hrs each burn wound was inoculated with 1x10^5 colony forming units (CFU) of GFP labelled Pseudomonas aeruginosa PAK. P. aeruginosa colonized and expanded within the tissue in a reproducible and time-dependent manner (n=8) and was assessed by quantitative culture after 24 and 48 hrs post-inoculation. Biofilm formation was determined visually by live-imaging with confocal microscopy.
Results: After 24 hrs individual cells in a patchy monolayer were found to preferentially attach to specific topographic features of the burn tissue. By 48 hrs post-infection sessile bacterial cells within highly ordered and three-dimensional structures up to 17 uM thick were observed. Multiple structures were observed across the surface of the burn wound and within the interstitial space where motile planktonic cells were found.
Conclusions: Together these data suggest that biofilm formation occurs non-uniformly in burn tissue in a manner that may be related to the degree of necrotic tissue. Future studies will focus on the evaluation of clinically-used wound treatments to determine their capacity for eradication of biofilm in burns.
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