Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (20): 3620-3625.doi: 10.3969/j.issn.1673-8225.2012.20.003

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Mineralization reaction during osteogenic differentiation of myoblasts stimulated by bone morphogenetic protein 2***☆

Zhang Li1, 2, Wang Wei2   

  1. 1Liaoning University of Traditional Chinese Medicine, Shenyang  110032, Liaoning Province, China; 2Department of Orthopedics, Jinzhou Central Hospital, Jinzhou  121000, Liaoning Province, China
  • Received:2011-11-03 Revised:2011-11-14 Online:2012-05-13 Published:2012-05-13
  • Contact: Wang Wei, Doctoral supervisor, Professor, Chief physician, Department of Orthopedics, Jinzhou Central Hospital, Jinzhou 121000, Liaoning Province, China weiwang_ly@yahoo.com.cn
  • About author:Zhang Li☆, Studying for doctorate, Attending physician, Liaoning University of Traditional Chinese Medicine, Shenyang 110032, Liaoning Province, China; Department of Orthopedics, Jinzhou Central Hospital, Jinzhou 121000, Liaoning Province, China zhangli2004jz@yahoo.com.cn
  • Supported by:

     the National Natural Science Foundation of China, No.30772190*; Postdoctorate Foundation Program of China, No. 2005038055*; Science and Technology Development Program of Liaoning Province, No. 2005225003-14*

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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.

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