Development of a High-Throughput ex-Vivo Burn Wound Model Using Porcine Skin, and Its Application to Evaluate New Approaches to Control Wound Infection

• Published in: Frontiers in cellular and infection microbiology Vol. 8; p. 196
• Main Authors: Alves, Diana R, Booth, Simon P, Scavone, Paola, Schellenberger, Pascale, Salvage, Jonathan, Dedi, Cinzia, Thet, Naing-Tun, Jenkins, A Toby A, Waters, Ryan, Ng, Keng W, Overall, Andrew D J, Metcalfe, Anthony D, Nzakizwanayo, Jonathan, Jones, Brian V
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 15.06.2018
• Subjects: accessory gene regulator; Animals; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; bacteriophage therapy; biofilm; Biofilms - growth & development; Burns - microbiology; Burns - pathology; Burns - veterinary; Cellular and Infection Microbiology; ex-vivo burn wound model; Humans; infection; Phage Therapy - veterinary; Reproducibility of Results; Skin - injuries; Skin - pathology; Staphylococcal Infections - pathology; Staphylococcal Infections - therapy; Staphylococcal Infections - veterinary; Staphylococcal Infections - virology; Staphylococcus aureus; Staphylococcus aureus - growth & development; Staphylococcus aureus - pathogenicity; Staphylococcus aureus - physiology; Staphylococcus aureus - virology; Swine; Trans-Activators - genetics; Trans-Activators - metabolism; Virulence Factors - genetics; Virulence Factors - physiology; Wound Infection - prevention & control; Wound Infection - therapy; Wound Infection - veterinary; Wound Infection - virology; infection; biofilm; bacteriophage therapy; infection responsive materials; ex-vivo burn wound model; Staphylococcus aureus; accessory gene regulator
• ISSN: 2235-2988; 2235-2988
• DOI: 10.3389/fcimb.2018.00196

Biofilm formation in wounds is considered a major barrier to successful treatment, and has been associated with the transition of wounds to a chronic non-healing state. Here, we present a novel laboratory model of wound biofilm formation using porcine skin and a custom burn wound array device. The model supports high-throughput studies of biofilm formation and is compatible with a range of established methods for monitoring bacterial growth, biofilm formation, and gene expression. We demonstrate the use of this model by evaluating the potential for bacteriophage to control biofilm formation by , and for population density dependant expression of virulence factors (regulated by the Accessory Gene Regulator, ) to signal clinically relevant wound infection. Enumeration of colony forming units and metabolic activity using the XTT assay, confirmed growth of bacteria in wounds and showed a significant reduction in viable cells after phage treatment. Confocal laser scanning microscopy confirmed the growth of biofilms in wounds, and showed phage treatment could significantly reduce the formation of these communities. Evaluation of activity by qRT-PCR showed an increase in activity during growth in wound models for most strains. Activation of a prototype infection-responsive dressing designed to provide a visual signal of wound infection, was related to increased activity. In all assays, excellent reproducibility was observed between replicates using this model.