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

Abstract

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.

• Authors: S Almqvist, M Werthén, A Johansson, J Törnqvist, M S Agren, P Thomsen
• Journal: The British journal of dermatology (Br J Dermatol) Vol. 160 Issue 6 Pg. 1163-71 (Jun 2009) ISSN: 1365-2133 [Electronic] England
• PMID: 19298284 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Amelogenin
• Chemokine CCL2
• Collagen Type I
• Interleukin-8
• Vascular Endothelial Growth Factor A

Topics

• Adult
• Aged, 80 and over
• Amelogenin (pharmacology)
• Area Under Curve
• Cell Line (drug effects, metabolism)
• Cell Proliferation (drug effects)
• Cells, Cultured
• Cellular Senescence (physiology)
• Chemokine CCL2 (biosynthesis)
• Collagen Type I (biosynthesis)
• Female
• Fibroblasts (drug effects, physiology)
• Humans
• Interleukin-8 (biosynthesis)
• Male
• Vascular Endothelial Growth Factor A (biosynthesis)