Effects of basic fibroblast growth factor on proliferation, migration, differentiation, and matrix production of human dental pulp cells
Date Issued
2010
Date
2010
Author(s)
Chang, Ya-Chin
Abstract
Aim: Basic fibroblast growth factor (bFGF) is multifunctional protein with a wide variety of effects. The functions of bFGF include wound healing, angiogenesis, melanogenesis, development of bone and cartilage, and so on. The purpose of our study is to investigate whether basic fibroblast growth factor influences the morphological changes, cell proliferation and viability, cell differentiation, and extracellular matrix formation of human dental pulp cells in vitro within the period of 5 days.
Materials and Methods: Primary-cultured human dental pulp cells were treated with different concentrations of bFGF (0, 1, 10, 50, 200 ng/ml). Morphology of pulp cells and cell migration were observed under light microscopy (40X). Cell proliferation was evaluated by MTT assay. Cell differentiation was evaluated by alkaline phosphatase (ALP) staining. Changes in mRNA expression of ALKP, Runx2, Cyclin B1, CDC2, CDC25c, p21, Rho, ROCK, a-sma, collagen type I, TIMP2, and MMP2 were determined by reverse-transcriptase polymerase chain reaction (RT-PCR). Collagen content was determined by Sircol Collagen assay.
Results: Dental pulp cells are spindle with extended cellular processes with/without bFGF treatment. The numbers of dental pulp cells without any bFGF treatment were much lower than those under the treatment of various concentrations of bFGF. Cell viability was two to three fold increased in higher concentrations of bFGF (50, 200 ng/ml). bFGF increased Cyclin B1, CDC2, and CDC25C mRNA gene expressions dose-dependently. Migrating rates were higher in the groups of higher concentrations of bFGF than those in the groups of lower concentrations. Rho and ROCK gene expressions were at peak at 10 ng/ml of bFGF. Nevertheless, α-sma mRNA gene expression of pulp cells declined in dose-dependent manner. bFGF downregulated ALPactivity of human dental pulp cells but did not affect significantly RUNX2 mRNA expression. bFGF did not affect significantly collagen content . Gene expression of MMP2 and TIMP2 were down-regulated by bFGF treatment.
Conclusion: Human dental pulp cells incubated with basic fibroblast growth factor demonstrated proliferative and migratory properties during the early stage of wound repairing (within 5 days). Meanwhile, bFGF had inverse effects on cell differentiation and extracellular matrix formation.
Subjects
dental pulp cells
bFGF
cell proliferation and migration
Type
thesis
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