Time- and dose-dependent effects of chronic wound fluid on human adult dermal fibroblasts.
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.
De Mattei
Monica
M
Department of Morphology and Embryology, Ferrara, Italy. dmm@unife.it
Ongaro
Alessia
A
Magaldi
Simona
S
Gemmati
Donato
D
Legnaro
Andrea
A
Palazzo
Annunziata
A
Masieri
Federica
F
Pellati
Agnese
A
Catozzi
Linda
L
Caruso
Angelo
A
Zamboni
Paolo
P
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