골수유래줄기세포에서 분화된 골유사세포에서 β-TCP와 rhBMP-2의 골형성 효과에 관한 연구
The effects of β-TCP/rhBMP-2 on bone formation in osteoblast-like cells induced from bone marrow-derived mesenchymal stem cells
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초록
The present study aimed to investigate the osteogenic potentials of differentiated osteoblast-like cells (DOCs) induced from bone marrow-derived mesenchymal stem cells (MSCs) on β-tricalcium phosphate (β-TCP) with recombinant human bone morphogenetic protein (rhBMP-2) in vitro. Osteoblast differentiation was induced in confluent cultures by adding 100 nM dexamethasone, 10 mM β-glycerophosphate, 50 mM L-ascorbic acid. The Alizarin red S staining and reverse transcriptase-polymerase chain reaction (RT-PCR) were perfomed to examine the mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), receptor activator for nuclear factor κB ligand (RANKL), runt-related transcription factor 2 (RUNX2), collagen-Ⅰ (COL-Ⅰ). There were no significant differences in the osteogenic potentials of DOCs induced from MSCs on β-TCP(+/-). According to the incubation period, there were significant increasing of Alizadin red S staining in the induction 3 weeks. The mRNA expression of ALP, RUNX2, and RANKL were higher in DOCs/β-TCP(-) than DOCs/β-TCP(+). According to rhBMP-2 concentrations, the mRNA expression of BSP was significantly increased in DOCs/β-TCP(+) compared to that of DOCs/β-TCP(-) on rhBMP 10 ng/ml. Our study presented the β-TCP will have the possibility that calcium phosphate directly affect the osteoblastic differentiation of the bone marrowderived MSCs.
Abstract
The present study aimed to investigate the osteogenic potentials of differentiated osteoblast-like cells (DOCs) induced from bone marrow-derived mesenchymal stem cells (MSCs) on β-tricalcium phosphate (β-TCP) with recombinant human bone morphogenetic protein (rhBMP-2) in vitro. Osteoblast differentiation was induced in confluent cultures by adding 100 nM dexamethasone, 10 mM β-glycerophosphate, 50 mM L-ascorbic acid. The Alizarin red S staining and reverse transcriptase-polymerase chain reaction (RT-PCR) were perfomed to examine the mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), receptor activator for nuclear factor κB ligand (RANKL), runt-related transcription factor 2 (RUNX2), collagen-Ⅰ (COL-Ⅰ). There were no significant differences in the osteogenic potentials of DOCs induced from MSCs on β-TCP(+/-). According to the incubation period, there were significant increasing of Alizadin red S staining in the induction 3 weeks. The mRNA expression of ALP, RUNX2, and RANKL were higher in DOCs/β-TCP(-) than DOCs/β-TCP(+). According to rhBMP-2 concentrations, the mRNA expression of BSP was significantly increased in DOCs/β-TCP(+) compared to that of DOCs/β-TCP(-) on rhBMP 10 ng/ml. Our study presented the β-TCP will have the possibility that calcium phosphate directly affect the osteoblastic differentiation of the bone marrowderived MSCs.
참고문헌 (38)
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