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Effects of FGF-2 on the Proliferation and Osteogenic Differentiation of the Adult Mesenchymal Stem Cells from Human Bone Marrow
Wen-Jie Tang1, 2, Ma-Lin Li1, Chui-Yuan Qiu2, Qiong-Yu Chen2, Guo-Hui Li, Ling-Song Li3, An Hong2*
1Yunnan Pharmacological Laboratory of Natural Products, Kunming Medical College, Kunming 650031, China;2Bioengineeering Institute of Jinan University, Guangzhou 510632, China;3The Centre of Stem Cell Research, University o
Abstract: To study the effects of fibroblast growth factor-2 (FGF-2) and/or dexamethasone (Dex) on the proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (MSCs) from passage 7 (P7) in vitro. Following the treatment with different mediums containing FGF-2 and/or Dex in vitro， the proliferation of human bone marrow MSCs (from P7) was evaluated via MTT assay; for osteogenic differentiation， the alkaline phosphatase (ALP) activity was determined by biochemical colorimetric assay with pNPP， and the contents of osteocalcin (OC) was detected by ELISA assay at different times， and then the assay of extracellular matrix mineralization was based on the detection of calcium mineral deposition using alizarin red S staining. When MSCs of P7 were cultured at low-cell-density in vitro， the growth rate of MSCs was 1.31-fold higher in the cultures treated with FGF-2 as compared to that of control at confluence. The growth rate of MSCs in the cultures treated with Dex/FGF-2 increased 1.47-fold or 1.12-fold respectively compared to that of control or FGF-2 treated cultures. The growth rate of MSCs did not alter obviously in the cultures treated with Dex alone. Under osteogenic differentiation culture conditions， the treatment with Dex increased the ALP activity (17.0-fold)， and OC contents (2.12-fold) of MSCs. Alizarin red S staining of cells indicated that Dex enhanced calcium mineral deposition in extracellular matrix (10.56-fold) and could mature mineral deposition into hydroxyapatite (HA) crystal and bone-like nodules. FGF-2 treatment decreased ALP activity (76.7%)， increased OC contents and calcium mineral deposition of MSCs. In the treatment of FGF-2 alone， the MSCs formed amorphous calcium mineral deposition and failed to mature into HA crystal and bone-like nodules. Thus， FGF-2 antagonized the induction of Dex on the ALP and HA crystal formation of MSCs. Results suggest that FGF-2 increases the proliferation potential of MSCs while Dex alone has not this effect. The combination of FGF-2 and Dex increases the proliferation potential at a level much higher than the FGF-2 alone. Dex can introduce MSCs into mature osteoblast as a potent osteogenic inducer. FGF-2 can also stimulates the osteogenic differentiation of MSCs， but the differentiated cells remain in immature state. FGF-2 antagonists the effect of inducing mature osteocytes by Dex.
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