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Evaluation of a near‐senescent human dermal fibroblast cell line and effect of amelogenin
Author(s) -
Almqvist S.,
Werthén M.,
Johansson A.,
Törnqvist J.,
Ågren M.S.,
Thomsen P.
Publication year - 2009
Publication title -
british journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.304
H-Index - 179
eISSN - 1365-2133
pISSN - 0007-0963
DOI - 10.1111/j.1365-2133.2009.09071.x
Subject(s) - amelogenin , fibroblast , extracellular matrix , dermal fibroblast , cell growth , cell culture , wound healing , type i collagen , microbiology and biotechnology , monocyte , chemistry , immunology , endocrinology , medicine , biology , biochemistry , gene , genetics
Summary Background Fibroblast senescence may delay healing of chronic wounds. Objectives To characterize a chronic human dermal fibroblast cell line (CRL‐7815) with near‐senescent properties, cell proliferation and production of wound‐healing modulating cytokines, and biosynthesis and remodelling of collagen were compared with normal human dermal fibroblasts. Also, the response of CRL‐7815 fibroblasts to the extracellular matrix protein amelogenin that is beneficial in the treatment of stalled chronic wounds was studied. Methods Fibroblast proliferation was monitored by time‐resolved growth curves and factors secreted into the culture medium containing 10% fetal bovine serum were measured by enzyme‐linked immunosorbent assays. Fibroblast‐mediated reorganization was examined in three‐dimensional type I collagen matrices. Results Cell proliferation over 9 days was significantly ( P < 0·01) slower for CRL‐7815 than for normal fibroblasts. Amelogenin at 1 mg mL −1 increased ( P < 0·01) CRL‐7815 proliferation to the level of the normal fibroblasts. The neutrophil chemoattractant interleukin (IL)‐8 was low while the constitutive production of monocyte chemoattractant protein (MCP)‐1 was highly elevated in medium from cultured CRL‐7815 fibroblasts. Amelogenin augmented IL‐8 but attenuated MCP‐1 secretion in CRL‐7815 fibroblasts. The elevated vascular endothelial growth factor production in CRL‐7815 fibroblasts was further increased with amelogenin while increased type I collagen synthesis by CRL‐7815 was reduced with 0·1 mg mL −1 amelogenin. The dramatically impaired collagen matrix remodelling with CRL‐7815 fibroblasts ( P < 0·001) was slightly improved with amelogenin ( P = 0·0011). Conclusions The near‐senescent cell line CRL‐7815 shares functional anomalies with fibroblasts isolated from nonhealing chronic cutaneous wounds. Amelogenin has the capacity to switch chronic fibroblasts into an acute‐like phenotype.