Mineralization reaction during osteogenic differentiation of myoblasts stimulated by bone morphogenetic protein 2***☆
- VernacularTitle:重组人骨形成蛋白2诱导成肌细胞成骨分化中的矿化反应
- Author:
Li ZHANG
;
Wei WANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2012;16(20):3620-3625
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: In recent years, it has been confirmed by a variety of ways that myoblasts can differentiate into osteoblasts under the induction of recombinant human bone morphogenetic protein 2 (rhBMP-2).OBJECTIVE: To explore the mineralization reaction during the osteogenic differentiation of myoblasts under the induction of recombinant rhBMP-2 and the feasibility of osteogenic phenotype expression by in vitro induction. METHODS: Myoblasts were isolated and harvested from neonatal Wistar rats using differential velocity adherent technique and trypsinization method. After in vitro culture, purification and identification, myoblasts at passage 3 were induced by a medium containing rhBMP-2 for 21 days. Myoblasts in the control group were cultured in vitro in complete medium without rhBMP-2 for 21 days. RESULTS AND CONCLUSION: After rhBMP-2 induction, myoblast proliferation gradually slowed down. A small quantity of opaque secretory granules were found in the cytoplasm on day 8 after induction; the number of opaque secretory granules increased on day 14 after induction; and a great quantity of opaque secretory granules were found in the cytoplasm on day 21 after induction while the myoblasts without induction fused into contractile myotubes. The alkaline phosphatase activity of the induced myoblasts increased as time extended; myoblasts reacted positively in the alkaline phosphatase staining, immunochemical staining for type Ⅰ collagen and calcium node staining on day 21 after induction. These findings suggest that mineralization reaction is found in rat myoblasts by rhBMP-2 induction and myoblasts can differentiate into osteoblasts under certain inducing conditions in vitro.
