Evaluation of a near-senescent human dermal fibroblast cell line and effect of amelogenin

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Abstract:

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.

Keywords: cell culture; enamel matrix derivative; extracellular matrix; wound healing

Document Type: Research Article

DOI: http://dx.doi.org/10.1111/j.1365-2133.2009.09071.x

Affiliations: 1: Department of Biomaterials, Sahlgrenska Academy at Göteborg University, Göteborg, Sweden 2: Mölnlycke Health Care AB, Göteborg, Sweden 3: Department of Surgery K and Copenhagen Wound Healing Center, Bispebjerg Hospital, Copenhagen NV, Denmark

Publication date: June 1, 2009

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