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Success rate of split-thickness skin grafting of chronic venous leg ulcers depends on the presence of Pseudomonas aeruginosa: a retrospective study.

Høgsberg T, Bjarnsholt T, Thomsen JS, Kirketerp-Møller K - PLoS ONE (2011)

Bottom Line: The results were evaluated using a Student T-test for continuous parameters, chi-square test for categorical parameters and a logistic regression analysis to predict healing after 12 weeks.Smoking also significantly suppressed the outcome at the 12-week follow-up.Subsequently, a logistic regression analysis was carried out leaving P. aeruginosa as the only predictor left in the model (p = 0.001).

View Article: PubMed Central - PubMed

Affiliation: Copenhagen Wound Healing Centre, Bispebjerg Hospital, Copenhagen, Denmark.

ABSTRACT
The last years of research have proposed that bacteria might be involved in and contribute to the lack of healing of chronic wounds. Especially it seems that Pseudomonas aeruginosa play a crucial role in the healing. At Copenhagen Wound Healing Centre it was for many years clinical suspected that once chronic venous leg ulcers were colonized (weeks or months preoperatively) by P. aeruginosa, the success rate of skin grafting deteriorated despite aggressive treatment. To investigate this, a retrospective study was performed on the clinical outcome of 82 consecutive patients with chronic venous leg ulcers on 91 extremities, from the 1(st) of March 2005 until the 31(st) of August 2006. This was achieved by analysing the microbiology, demographic data, smoking and drinking habits, diabetes, renal impairment, co-morbidities, approximated size and age of the wounds, immunosuppressive treatment and complicating factors on the clinical outcome of each patient. The results were evaluated using a Student T-test for continuous parameters, chi-square test for categorical parameters and a logistic regression analysis to predict healing after 12 weeks. The analysis revealed that only 33,3% of ulcers with P. aeruginosa, isolated at least once from 12 weeks prior, to or during surgery, were healed (98% or more) by week 12 follow-up, while 73,1% of ulcers without P. aeruginosa were so by the same time (p = 0.001). Smoking also significantly suppressed the outcome at the 12-week follow-up. Subsequently, a logistic regression analysis was carried out leaving P. aeruginosa as the only predictor left in the model (p = 0.001). This study supports our hypothesis that P. aeruginosa in chronic venous leg ulcers, despite treatment, has considerable impact on partial take or rejection of split-thickness skin grafts.

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CLSM three-dimensional image of a large P. aeruginosa biofilm in the wound bed of a chronic wound (identified by a specific PNA FISH probe (red)).
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pone-0020492-g003: CLSM three-dimensional image of a large P. aeruginosa biofilm in the wound bed of a chronic wound (identified by a specific PNA FISH probe (red)).

Mentions: Tissue section of a chronic wound was analyzed by fluorescence in situ hybridization (FISH) using peptide nucleic acid (PNA) probes (Figure 3). A Texas Red-labeled P. aeruginosa-specific PNA probe in hybridization solution (AdvanDx, Inc., Woburn, MA), was added to each section and hybridized in a PNA FISH workstation covered by a lid at 55°C for 90 min. The slides were washed for 30 min at 55°C in wash solution (AdvanDx). Three-dimensional imaging of the bacteria in the wounds was done by confocal laser scanning microscopy (CLSM) using a Zeiss LSM 510 system (Carl Zeiss GmbH, Jena, Germany) equipped with ahelium-neon laser for excitation of the fluorophores. Simulated fluorescence projections of the biofilms were generated by using the IMARIS software package (Bitplane AG). Images were further processed for display by using Photoshop software (Adobe).


Success rate of split-thickness skin grafting of chronic venous leg ulcers depends on the presence of Pseudomonas aeruginosa: a retrospective study.

Høgsberg T, Bjarnsholt T, Thomsen JS, Kirketerp-Møller K - PLoS ONE (2011)

CLSM three-dimensional image of a large P. aeruginosa biofilm in the wound bed of a chronic wound (identified by a specific PNA FISH probe (red)).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3105064&req=5

pone-0020492-g003: CLSM three-dimensional image of a large P. aeruginosa biofilm in the wound bed of a chronic wound (identified by a specific PNA FISH probe (red)).
Mentions: Tissue section of a chronic wound was analyzed by fluorescence in situ hybridization (FISH) using peptide nucleic acid (PNA) probes (Figure 3). A Texas Red-labeled P. aeruginosa-specific PNA probe in hybridization solution (AdvanDx, Inc., Woburn, MA), was added to each section and hybridized in a PNA FISH workstation covered by a lid at 55°C for 90 min. The slides were washed for 30 min at 55°C in wash solution (AdvanDx). Three-dimensional imaging of the bacteria in the wounds was done by confocal laser scanning microscopy (CLSM) using a Zeiss LSM 510 system (Carl Zeiss GmbH, Jena, Germany) equipped with ahelium-neon laser for excitation of the fluorophores. Simulated fluorescence projections of the biofilms were generated by using the IMARIS software package (Bitplane AG). Images were further processed for display by using Photoshop software (Adobe).

Bottom Line: The results were evaluated using a Student T-test for continuous parameters, chi-square test for categorical parameters and a logistic regression analysis to predict healing after 12 weeks.Smoking also significantly suppressed the outcome at the 12-week follow-up.Subsequently, a logistic regression analysis was carried out leaving P. aeruginosa as the only predictor left in the model (p = 0.001).

View Article: PubMed Central - PubMed

Affiliation: Copenhagen Wound Healing Centre, Bispebjerg Hospital, Copenhagen, Denmark.

ABSTRACT
The last years of research have proposed that bacteria might be involved in and contribute to the lack of healing of chronic wounds. Especially it seems that Pseudomonas aeruginosa play a crucial role in the healing. At Copenhagen Wound Healing Centre it was for many years clinical suspected that once chronic venous leg ulcers were colonized (weeks or months preoperatively) by P. aeruginosa, the success rate of skin grafting deteriorated despite aggressive treatment. To investigate this, a retrospective study was performed on the clinical outcome of 82 consecutive patients with chronic venous leg ulcers on 91 extremities, from the 1(st) of March 2005 until the 31(st) of August 2006. This was achieved by analysing the microbiology, demographic data, smoking and drinking habits, diabetes, renal impairment, co-morbidities, approximated size and age of the wounds, immunosuppressive treatment and complicating factors on the clinical outcome of each patient. The results were evaluated using a Student T-test for continuous parameters, chi-square test for categorical parameters and a logistic regression analysis to predict healing after 12 weeks. The analysis revealed that only 33,3% of ulcers with P. aeruginosa, isolated at least once from 12 weeks prior, to or during surgery, were healed (98% or more) by week 12 follow-up, while 73,1% of ulcers without P. aeruginosa were so by the same time (p = 0.001). Smoking also significantly suppressed the outcome at the 12-week follow-up. Subsequently, a logistic regression analysis was carried out leaving P. aeruginosa as the only predictor left in the model (p = 0.001). This study supports our hypothesis that P. aeruginosa in chronic venous leg ulcers, despite treatment, has considerable impact on partial take or rejection of split-thickness skin grafts.

Show MeSH
Related in: MedlinePlus