Recombinant human bone morphogenic protein-2 Induces the Differentiation and Mineralization of Osteoblastic Cells Under Hypoxic Conditions via Activation of Protein Kinase D and p38 Mitogen-Activated Protein Kinase Signaling Pathways.
10.1007/s13770-017-0046-1
- Author:
Woo Hun HA
1
;
Hwa Sik SEONG
;
Na Rae CHOI
;
Bong Soo PARK
;
Yong Deok KIM
Author Information
1. Department of Oral and Maxillofacial Surgery, Pusan National University, Beomeori, Mulgeom, Yangsan 50612, Korea. ydkimdds@pusan.ac.kr
- Publication Type:Original Article
- Keywords:
Hypoxia;
Pathway;
p38 MAPK;
PKD;
Inhibitor
- MeSH:
Alkaline Phosphatase;
Anoxia;
Blotting, Western;
Collagen;
Humans*;
Miners*;
Osteoblasts*;
Osteocalcin;
Oxygen;
p38 Mitogen-Activated Protein Kinases;
Phosphorylation;
Protein Kinases*;
Regenerative Medicine
- From:
Tissue Engineering and Regenerative Medicine
2017;14(4):433-441
- CountryRepublic of Korea
- Language:English
-
Abstract:
Hypoxia suppresses osteoblastic differentiation and the bone-forming capacity. As the leading osteoinductive growth factor used clinically in bone-related regenerative medicine, recombinant human bone morphogenic protein-2 (rhBMP- 2) has yielded promising results in unfavorable hypoxic clinical situations. Although many studies have examined the effects of rhBMP-2 on osteoblastic differentiation, mineralization and the related signaling pathways, those of rhBMP-2 on osteoblastic cells remain unknown, particularly under hypoxic conditions. Therefore, this study was conducted under a 1% oxygen tension to examine the differentiating effects of rhBMP-2 on osteoblastic cells under hypoxia. rhBMP-2 could also induce the differentiation and mineralization of Osteoblastic (MC3T3-E1) cells under1%hypoxic conditions. rhBMP-2 could also induce the differentiation and mineralization of MC3T3-E1 cells under 1% hypoxic conditions. rhBMP-2 increased the alkaline phosphatase {ALP} activity in a time dependent manner, and expression of ALP, collagen type-1 (Col-1) and osteocalcin (OC) mRNAwere up-regulated significantly in a time- and concentration-dependent manner. In addition, the area of the mineralized nodules increased gradually in a concentration-dependent manner. Western blot analysis, which was performed to identify the signaling pathways underlying rhBMP-2-induced osteoblastic differentiation under hypoxic conditions, showed that rhBMP-2 significantly promoted the phosphorylation of the p38 mitogen-activated protein kinase (MAPK) in a time-dependent manner. A pretreatment with SB203580, a p38 MAPK inhibitor, inhibited the rhBMP-2-mediated differentiation and mineralization. Moreover, the phosphorylation of p38 induced by rhBMP-2 was inhibited in response to a pretreatment of the cells with Go6976, a protein kinase D {PKD) inhibitor. These findings suggest that rhBMP-2 induces the differentiation and mineralization of MC3T3-E1 cells under hypoxic conditions via activation of the PKD and p38 MAPK signaling pathways.
