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 Ribeiro Alves, Simon Booth, Paola Scavone, Pascale Schellenberger, Jonathan Salvage, Cinzia Dedi, Naing-Tun Thet, Toby A. Jenkins, Ryan Waters, Keng W. Ng, Andrew Overall, Anthony Metcalfe, Jonathan Nzakizwanayo, Brian Jones

Research output: Contribution to journalArticleResearchpeer-review

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
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

Bibliographical note

© 2018 Alves, Booth, Scavone, Schellenberger, Salvage, Dedi, Thet, Jenkins, Waters, Ng, Overall, Metcalfe, Nzakizwanayo and Jones. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Cite this

Ribeiro Alves, Diana ; Booth, Simon ; Scavone, Paola ; Schellenberger, Pascale ; Salvage, Jonathan ; Dedi, Cinzia ; Thet, Naing-Tun ; Jenkins, Toby A. ; Waters, Ryan ; Ng, Keng W. ; Overall, Andrew ; Metcalfe, Anthony ; Nzakizwanayo, Jonathan ; Jones, Brian. / 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.
@article{0c8586c0536349a0a64cab21e4632901,
title = "Development of a High-Throughput ex-Vivo Burn Wound Model Using Porcine Skin, and Its Application to Evaluate New Approaches to Control Wound Infection",
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.",
author = "{Ribeiro Alves}, Diana and Simon Booth and Paola Scavone and Pascale Schellenberger and Jonathan Salvage and Cinzia Dedi and Naing-Tun Thet and Jenkins, {Toby A.} and Ryan Waters and Ng, {Keng W.} and Andrew Overall and Anthony Metcalfe and Jonathan Nzakizwanayo and Brian Jones",
note = "{\circledC} 2018 Alves, Booth, Scavone, Schellenberger, Salvage, Dedi, Thet, Jenkins, Waters, Ng, Overall, Metcalfe, Nzakizwanayo and Jones. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.",
year = "2018",
month = "6",
day = "15",
doi = "10.3389/fcimb.2018.00196",
language = "English",
journal = "Frontiers in Cellular and Infection Microbiology",

}

Development of a High-Throughput ex-Vivo Burn Wound Model Using Porcine Skin, and Its Application to Evaluate New Approaches to Control Wound Infection. / Ribeiro Alves, Diana; Booth, Simon; Scavone, Paola; Schellenberger, Pascale; Salvage, Jonathan; Dedi, Cinzia; Thet, Naing-Tun; Jenkins, Toby A.; Waters, Ryan; Ng, Keng W. ; Overall, Andrew; Metcalfe, Anthony; Nzakizwanayo, Jonathan; Jones, Brian.

In: Frontiers in Cellular and Infection Microbiology, 15.06.2018.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Development of a High-Throughput ex-Vivo Burn Wound Model Using Porcine Skin, and Its Application to Evaluate New Approaches to Control Wound Infection

AU - Ribeiro Alves, Diana

AU - Booth, Simon

AU - Scavone, Paola

AU - Schellenberger, Pascale

AU - Salvage, Jonathan

AU - Dedi, Cinzia

AU - Thet, Naing-Tun

AU - Jenkins, Toby A.

AU - Waters, Ryan

AU - Ng, Keng W.

AU - Overall, Andrew

AU - Metcalfe, Anthony

AU - Nzakizwanayo, Jonathan

AU - Jones, Brian

N1 - © 2018 Alves, Booth, Scavone, Schellenberger, Salvage, Dedi, Thet, Jenkins, Waters, Ng, Overall, Metcalfe, Nzakizwanayo and Jones. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PY - 2018/6/15

Y1 - 2018/6/15

N2 - 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.

AB - 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.

U2 - 10.3389/fcimb.2018.00196

DO - 10.3389/fcimb.2018.00196

M3 - Article

JO - Frontiers in Cellular and Infection Microbiology

JF - Frontiers in Cellular and Infection Microbiology

M1 - 196

ER -