Development of a high-throughput ex-vivo burn wound model using porcine skin, and its application to evaluate new approaches to control wound infection

Diana R. Alves, Simon P. Booth, Paola Scavone, Pascale Schellenberger, Jonathan Salvage, Cinzia Dedi, Naing Tun Thet, A. Toby A. Jenkins, Ryan Waters, Keng W. Ng, Andrew D.J. Overall, Anthony D. Metcalfe, Jonathan Nzakizwanayo, Brian V. Jones

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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 ex-vivo 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 Staphylococcus aureus, and for population density dependant expression of S. aureus virulence factors (regulated by the Accessory Gene Regulator, agr) 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 agr 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 agr activity. In all assays, excellent reproducibility was observed between replicates using this model.

Original languageEnglish
Article number196
JournalFrontiers in Cellular and Infection Microbiology
Volume8
Issue numberJUN
DOIs
Publication statusPublished - 15 Jun 2018

Fingerprint

Wound Infection
Biofilms
Swine
Skin
Wounds and Injuries
Regulator Genes
Bacteriophages
Growth
Staphylococcus aureus
Virulence Factors
Bandages
Population Density
Confocal Microscopy
Stem Cells
Bacteria
Gene Expression
Equipment and Supplies
Polymerase Chain Reaction
Infection

Keywords

  • Accessory gene regulator
  • Bacteriophage therapy
  • Biofilm
  • Ex-vivo burn wound model
  • Infection
  • Infection responsive materials
  • Staphylococcus aureus

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

Cite this

Development of a high-throughput ex-vivo burn wound model using porcine skin, and its application to evaluate new approaches to control wound infection. / 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.

In: Frontiers in Cellular and Infection Microbiology, Vol. 8, No. JUN, 196, 15.06.2018.

Research output: Contribution to journalArticle

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. / Development of a high-throughput ex-vivo burn wound model using porcine skin, and its application to evaluate new approaches to control wound infection. In: Frontiers in Cellular and Infection Microbiology. 2018 ; Vol. 8, No. JUN.
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