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 language | English |
|---|---|
| Article number | 196 |
| Journal | Frontiers in Cellular and Infection Microbiology |
| Volume | 8 |
| Issue number | JUN |
| DOIs | |
| Publication status | Published - 15 Jun 2018 |
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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 journal › Article
}
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 - Alves, Diana R.
AU - Booth, Simon P.
AU - Scavone, Paola
AU - Schellenberger, Pascale
AU - Salvage, Jonathan
AU - Dedi, Cinzia
AU - Thet, Naing Tun
AU - Jenkins, A. Toby A.
AU - Waters, Ryan
AU - Ng, Keng W.
AU - Overall, Andrew D.J.
AU - Metcalfe, Anthony D.
AU - Nzakizwanayo, Jonathan
AU - Jones, Brian V.
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.
KW - Accessory gene regulator
KW - Bacteriophage therapy
KW - Biofilm
KW - Ex-vivo burn wound model
KW - Infection
KW - Infection responsive materials
KW - Staphylococcus aureus
UR - http://www.scopus.com/inward/record.url?scp=85048602473&partnerID=8YFLogxK
U2 - 10.3389/fcimb.2018.00196
DO - 10.3389/fcimb.2018.00196
M3 - Article
AN - SCOPUS:85048602473
VL - 8
JO - Frontiers in Cellular and Infection Microbiology
JF - Frontiers in Cellular and Infection Microbiology
SN - 2235-2988
IS - JUN
M1 - 196
ER -