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Time- and Dose-Dependent Effects of Chronic Wound Fluid on Human Adult Dermal Fibroblasts |
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| Authors: | MONICA DE MATTEI PHD ALESSIA ONGARO PHD SIMONA MAGALDI BS DONATO GEMMATI BS REA LEGNARO MD ANNUNZIATA PALAZZO MD FEDERICA MASIERI BS AGNESE PELLATI BS LINDA CATOZZI BS ANGELO CARUSO PHD PAOLO ZAMBONI MD |
| Affiliation: | Department of Morphology and Embryology, Ferrara, Italy,;Centre for the Study of Haemostasis and Thrombosis, Ferrara, Italy,;Vascular Diseases Centre, University of Ferrara, Ferrara, Italy |
| 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. OBJECTIVES 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 μg/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, α-smooth muscle actin expression by immunofluorescence, and senescence-associated β-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. |
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