title: Time- and dose-dependent effects of chronic wound fluid on human adult dermal fibroblasts.
abstract: BACKGROUND
  Wound healing is a biologic process that is altered in patients affected by chronic venous ulcers. The wound microenvironment is reflected in the chronic wound fluid (CWF), an exudate containing serum components and tissue-derived proteins.
  
  
  OBJECTIVE
  We investigated the effects of increasing doses of CWF collected from patients suffering from chronic venous ulcers on human adult dermal fibroblasts cultured in vitro and the relationship among CWF effects and treatment length.
  
  
  METHODS
  Fibroblasts were treated with 60, 240, and 720 microg/mL CWF for 3 and 7 days. We evaluated cell proliferation and viability by MTT and Trypan blue assay, cell morphology by light microscopy, F-actin microfilaments organization by tetramethylrhodamine B isothiocyanate-conjugated phalloidin, alpha-smooth muscle actin expression by immunofluorescence, and senescence-associated beta-galactosidase activity.
  
  
  RESULTS
  CWF induced an increase in cell proliferation in the first 3 days of treatment. In contrast, at 7 days, a strong decrease in cell viability was observed. These changes were related to a cytoskeletal F-actin reorganization and not to fibroblast-myofibroblast differentiation nor to changes in cellular senescence.
  
  
  CONCLUSIONS
  This study shows a dose-dependent and biphasic effect of CWF on dermal fibroblasts, suggesting that a continuous exposure to chronic wounds microenvironment may induce late cellular dysfunctions possibly involved in the delayed wound healing.
authors.affiliation: Department of Morphology and Embryology, Ferrara, Italy. dmm@unife.it
authors.forename: Monica
authors.initials: M
authors.lastname: De Mattei
authors.forename: Alessia
authors.initials: A
authors.lastname: Ongaro
authors.forename: Simona
authors.initials: S
authors.lastname: Magaldi
authors.forename: Donato
authors.initials: D
authors.lastname: Gemmati
authors.forename: Andrea
authors.initials: A
authors.lastname: Legnaro
authors.forename: Annunziata
authors.initials: A
authors.lastname: Palazzo
authors.forename: Federica
authors.initials: F
authors.lastname: Masieri
authors.forename: Agnese
authors.initials: A
authors.lastname: Pellati
authors.forename: Linda
authors.initials: L
authors.lastname: Catozzi
authors.forename: Angelo
authors.initials: A
authors.lastname: Caruso
authors.forename: Paolo
authors.initials: P
authors.lastname: Zamboni
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D002908:Chronic Disease; D020405:Dermis / Q000473:pathology* / Q000503:physiopathology; D005122:Exudates and Transudates; D005347:Fibroblasts / Q000473:pathology*; D006801:Humans; D008875:Middle Aged; D013997:Time Factors; D014647:Varicose Ulcer / Q000503:physiopathology; D014945:Wound Healing / Q000502:physiology*; D014947:Wounds and Injuries / Q000473:pathology* / Q000503:physiopathology