Function of EGF on MCP-1 in cultured dental follicle cells

Objective: To study the effects of epidermal growth factor (EGF) on the secretion of monocyte chemotactic protein-one (MCP-1) in rat dental follicle cells. Methods: (1) Rat dental follicle cells culture was established, and the third passage cell was used in this study. To assess the effects of EGF on cell growth, 2x104cells were plated into 96 well microplates. After 24h in a-MEM containing 15% fetal bovine serum(FBS), the cells in each well were rinsed thrice ina-MEM without FBS. Controls were then cultured ina-MEM containing 0.5% serum, while experimental cells were plus additional different concentrations (0.5, 1, 5, 10, 50ng/ml) of EGF for 5 days. MTT assays were used to evaluate their proliferation in this time point. (2) 1x105 cells were plated into each 70 mm dia culture dish ina-MEM containing 0.5% serum for 5 days, the cells were cultured ina-MEM for 5 hr, and then the cells were incubated with 10ng/ml EGF(the prime effective concentration of EGF on dental follicle cells ) for 0, 0.5, 1, 3, 6h separately to determine its time-dependent effect on MCP-1 gene expression. The total RNA was then extracted from each sample for Reverse Transcription-Polymerase Chain Reaction analysis. Results: The cultured dental follicle cells were significantly proliferated under the challenge of EGF at the concentration of l-10ng/ml (P<0.05), and the prime effective concentration of EGF on dental follicle cells was 10ng/ml. Epidermal growth factor had a dramatic effect on MCP-1 expression, resulting in peak at 3 hr after incubation (P <0.01) with a decline at later times. Conclusion: Proper concentrations of epidermal growth factor may promote dental follicle cell proliferation, and it may enhance MCP-1 gene transcription in vitro.