Ex situ model of biofilm‐associated wounds: providing a host‐like environment for the study of Staphylococcus aureus and Pseudomonas aeruginosa biofilms

Aim This study aimed to assess an ex situ model of biofilm-associated wounds on porcine skin for the study of Staphylococcus aureus and Pseudomonas aeruginosa biofilms in a host-like environment, after 48 to 120 h of incubation. Material and results Ex situ and in vitro biofilms were comparatively analysed. Overall, CFU-counts and matrix quantification yielded significantly (P < 0·05) higher results for ex situ than in vitro biofilms. Confocal microscopy revealed greater (P < 0·05) biomass and thickness at 48–72 h and greater (P < 0·05) robustness at 72 h of growth. S. aureus ex situ biofilms produced less (P < 0·05) siderophore and proteases than in vitro biofilms, while P. aeruginosa ex situ biofilms produced more (P < 0·05) siderophores and less proteases than in vitro biofilms. Conclusions Biofilms grown ex situ present a greater amount of bacterial cells and polymeric matrix than their in vitro counterparts, reaching maturity at 72 h of growth. Moreover the production of virulence factors differs between ex situ and in vitro biofilms. Significance and Impact of the Study These findings emphasize the importance of using ex situ biofilm models, once they mimic in vivo conditions. The use of these models brings perspectives for the pursuit of therapeutic alternatives, as tests may be performed in a host-like environment.