title: Quantitative analysis of the cellular inflammatory response against biofilm bacteria in chronic wounds.
abstract: Chronic wounds are an important problem worldwide. These wounds are characterized by a persistent inflammatory stage associated with excessive accumulation and elevated cell activity of neutrophils, suggesting that there must be a persistent stimulus that attracts and recruits neutrophils to the wound. One such stimulus might be the presence of bacterial biofilms in chronic wounds. In the present study, biopsy specimens from chronic venous leg ulcers were investigated for the detection of bacteria using peptide nucleic acid-based fluorescence in situ hybridization (PNA-FISH) and confocal laser scanning microscopy. The bacteria in the wounds were often situated in large aggregates. To obtain a measure of the cellular inflammatory response against the bacteria in the chronic wounds, the amount of neutrophils accumulated at the site of infection was evaluated through differential neutrophil counting on the tissue sections from wounds containing either Pseudomonas aeruginosa or Staphylococcus aureus. The P. aeruginosa-containing wounds had significantly higher numbers of neutrophils accumulated compared with the S. aureus-containing wounds. These results are discussed in relation to the hypothesis that the presence of P. aeruginosa biofilms in chronic wounds may be one of the main factors leading to a persistent inflammatory response and impaired wound healing.
authors.affiliation: Department of International Health, Immunology and Microbiology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Fazli
authors.forename: Thomas
authors.initials: T
authors.lastname: Bjarnsholt
authors.forename: Klaus
authors.initials: K
authors.lastname: Kirketerp-Møller
authors.forename: Anne
authors.initials: A
authors.lastname: Jørgensen
authors.forename: Claus Bøgelund
authors.initials: CB
authors.lastname: Andersen
authors.forename: Michael
authors.initials: M
authors.lastname: Givskov
authors.forename: Tim
authors.initials: T
authors.lastname: Tolker-Nielsen
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D018441:Biofilms; D002908:Chronic Disease; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007249:Inflammation / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D020556:Neutrophil Infiltration; D020135:Peptide Nucleic Acids; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification / Q000502:physiology; D013211:Staphylococcus aureus / Q000302:isolation & purification / Q000502:physiology; D014647:Varicose Ulcer / Q000378:metabolism* / Q000382:microbiology*; D014945:Wound Healing / Q000276:immunology / Q000502:physiology*; D014946:Wound Infection / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology*
chemical_list: D020135:Peptide Nucleic Acids