Negative-Pressure Wound Therapy in a Pseudomonas aeruginosa Infection Model

BACKGROUND: Negative-pressure wound therapy (NPWT) is an effective strategy for the management of contaminated wounds, including those infected by Pseudomonas aeruginosa. We hypothesized that NPWT would reduce virulence factors as well as biofilm components and inhibit virulence-regulated gene expression in a model of P. aeruginosa wound infection. METHODS: Wounds were created in anesthetized rabbits and P. aeruginosa was inoculated to the wound surface for 24 h. Wounds were treated with either NPWT or a sterile gauze dressing. Virulence factors including exotoxin A, rhamnolipid, and elastase were quantified by the enzyme-linked immunosorbent assay, orcinol, and elastin-Congo red methods, respectively. A biofilm component, eDNA, was quantified using a commercial kit. Virulence-regulated genes were determined by quantitative real-time polymerase chain reaction (RT-PCR). Biofilms were observed in vivo by staining with concanavalin A conjugated to Alexa Fluor® 647. RESULTS: NPWT was more effective than the control treatment in reducing virulence factors and bacteria counts in vivo. A biofilm component, eDNA, was less abundant in the NPWT group. The results of the RT-PCR indicated that the expression levels of P. aeruginosa virulence-regulated genes and quorum-sensing population density-dependent systems were significantly inhibited by NPWT treatment. CONCLUSION: NPWT reduced bacteria counts, virulence factors, and eDNA in a P. aeruginosa wound infection model in vivo. These beneficial effects are likely to be related to the reduced expression of virulence-regulated genes and the drainage induced by NPWT treatment. These findings may help clinicians to obtain a better understanding of the mechanism of NPWT for the treatment of infected wounds.