title: Proteolytic activity by multiple bacterial species isolated from chronic venous leg ulcers degrades matrix substrates.
abstract: BACKGROUND
  A major feature of chronic wounds is the loss of tissue, with the exposure of dermal components preventing primary closure and leading to bacterial colonization. Bacterial colonization has been proposed as one of the common underlying pathologies present in chronic wounds. The objective of this exploratory study was to identify bacteria cultured from chronic venous leg ulcers and test for proteolytic activity that degrades matrix substrates.
  
  
  METHODS
  Bacteria were isolated, cultured, and identified from six subjects (average age = 62.8 years) over 2-10 months under an approved protocol using swabs and microbiological culture media. Proteolytic activity against (a) gelatin, (b) an elastin substrate, and (c) a serine/trypsin-sensitive substrate was determined using a colorimetric plate assay with an ELISA plate reader and zymography.
  
  
  RESULTS
  We identified 13 bacteria that expressed proteolytic activity against one or more of the tested substrates. Of these, six were Gram-positive (Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Streptococcus agalactiae, Corynebacterium, and Streptococcus bovis) and seven were Gram-negative (Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Morganella morganii, Klebsiella pneumoniae, Bacteroides fragilis, and Serratia marcescens) organisms. Two of these, S. aureus and P. aeruginosa, are recognized wound pathogens.
  
  
  CONCLUSIONS
  Multiple bacteria species isolated from colonized venous leg ulcers have the capacity to secrete proteases capable of degrading components of the extracellular matrix important for wound healing. Matrix degradation by bacteria may contribute to delays in tissue deposition and repair, suggesting that treatment of chronic wounds should include appropriate management of colonizing bacteria.
authors.affiliation: 1School of Nursing, University of Massachusetts Amherst, Amherst, MA, USA.
authors.forename: Annette B
authors.initials: AB
authors.lastname: Wysocki
authors.forename: Sandhya K
authors.initials: SK
authors.lastname: Bhalla-Regev
authors.forename: Philip M
authors.initials: PM
authors.lastname: Tierno
authors.forename: Marla
authors.initials: M
authors.lastname: Stevens-Riley
authors.forename: Ryan-Claire
authors.initials: RC
authors.lastname: Wiygul
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002908:Chronic Disease; D004549:Elastin / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D005780:Gelatin / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000378:metabolism*; D006094:Gram-Positive Bacteria / Q000378:metabolism*; D006097:Granulation Tissue / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D007871:Leg Ulcer / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D008297:Male; D008828:Microbiological Techniques; D008875:Middle Aged; D059748:Proteolysis; D012867:Skin / Q000382:microbiology*
chemical_list: D005780:Gelatin; D004549:Elastin
keywords: bacteria; extracellular matrix; leg ulcer; protease; wound