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Bone morphogenetic protein receptor in the osteogenic differentiation of rat bone marrow stromal cells.

Wang A, Ding X, Sheng S, Yao Z - Yonsei Med. J. (2010)

Bottom Line: Bone morphogenetic protein (BMP) signaling has important effects on the process of skeletogenesis.The OM significantly induced the expression of type IA receptor of BMPR (BMPRIA) and type II receptor of BMPR (BMPRII) in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively.In addition, BMP signaling through BMPRIA and BMPRII regulates the osteogenic differentiation of rat BMSCs in OM with or without BMP-2.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital, Sun Yat-sen University, 58 Zhongshan Road II, Guangzhou, 510080, PR China. anxunwang@yahoo.com

ABSTRACT

Purpose: Several signaling pathways have been shown to regulate the lineage commitment and terminal differentiation of bone marrow stromal cells (BMSCs). Bone morphogenetic protein (BMP) signaling has important effects on the process of skeletogenesis. In the present study, we tested the role of bone morphogenetic protein receptor (BMPR) in the osteogenic differentiation of rat bone marrow stromal cells in osteogenic medium (OM) with or without BMP-2.

Materials and methods: BMSCs were harvested from rats and cultured in OM containing dexamethasone, beta-glycerophosphate, and ascorbic acid, with or without BMP-2 in order to induce osteogenic differentiation. The alkaline phosphatase (ALP) activity assay and von kossa staining were used to assess the osteogenic differentiation of the BMSCs. BMPR mRNA expression was assessed using reverse transcriptionpolymerase chain reaction (RT-PCR).

Results: The BMSCs that underwent osteogenic differentiation in OM showed a higher level of ALP activity and matrix mineralization. BMP-2 alone induced a low level of ALP activity and matrix mineralization in BMSCs, but enhanced the osteogenic differentiation of BMSCs when combined with OM. The OM significantly induced the expression of type IA receptor of BMPR (BMPRIA) and type II receptor of BMPR (BMPRII) in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively.

Conclusion: BMSCs commit to osteoblastic differentiation in OM, which is enhanced by BMP-2. In addition, BMP signaling through BMPRIA and BMPRII regulates the osteogenic differentiation of rat BMSCs in OM with or without BMP-2.

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mRNA expression of the BMP receptor in BMSCs. BMSCs were treated with CM or OM with or without BMP-2. mRNA was extracted from cells on day 3, 6, and 9 and analyzed by RT-PCR. GADPH mRNA was used as an internal control to normalize the amount of RNA. The relative expression level of mRNA was depicted as the ratio of the density of mRNA to GADPH mRNA at the same time point. The results correspond to a representative of three experiments. RT-PCR analysis revealed that the osteogenic medium significantly induced BMPR-IA and BMPRII expression in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively. *Compared with CM at the same time point, p < 0.05. †Compared with BMP-2 at the same time point, p < 0.05. CM, control medium; OM, osteogenic medium; BMP-2, bone morphogenetic protein-2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; BMPRIA, type IA receptor of BMPR; BMPRII, type II receptor of BMPR; RT-PCR, reverse transcription-polymerase chain reaction; BMSCs, bone marrow stromal cells.
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Figure 4: mRNA expression of the BMP receptor in BMSCs. BMSCs were treated with CM or OM with or without BMP-2. mRNA was extracted from cells on day 3, 6, and 9 and analyzed by RT-PCR. GADPH mRNA was used as an internal control to normalize the amount of RNA. The relative expression level of mRNA was depicted as the ratio of the density of mRNA to GADPH mRNA at the same time point. The results correspond to a representative of three experiments. RT-PCR analysis revealed that the osteogenic medium significantly induced BMPR-IA and BMPRII expression in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively. *Compared with CM at the same time point, p < 0.05. †Compared with BMP-2 at the same time point, p < 0.05. CM, control medium; OM, osteogenic medium; BMP-2, bone morphogenetic protein-2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; BMPRIA, type IA receptor of BMPR; BMPRII, type II receptor of BMPR; RT-PCR, reverse transcription-polymerase chain reaction; BMSCs, bone marrow stromal cells.

Mentions: Because BMP signaling has been shown to promote the osteoblastic differentiation of BMSCs, BMP receptors were assessed by RT-PCR during BMSC differentiation. As shown in Fig. 4, the expression levels of BMPR-IA in OM with or without BMP-2 were significantly higher than CM or BMP-2 at three or six days of stimulation (p < 0.05). No difference was found between the CM and BMP-2 groups. Nine days after stimulation, the level of BMPRIA mRNA was continuously induced in OM with or without BMP-2 (p < 0.05). The expression of BMPRIA significantly increased in the BMP-2 groups as compared to the other three groups at nine days of stimulation (p < 0.05).


Bone morphogenetic protein receptor in the osteogenic differentiation of rat bone marrow stromal cells.

Wang A, Ding X, Sheng S, Yao Z - Yonsei Med. J. (2010)

mRNA expression of the BMP receptor in BMSCs. BMSCs were treated with CM or OM with or without BMP-2. mRNA was extracted from cells on day 3, 6, and 9 and analyzed by RT-PCR. GADPH mRNA was used as an internal control to normalize the amount of RNA. The relative expression level of mRNA was depicted as the ratio of the density of mRNA to GADPH mRNA at the same time point. The results correspond to a representative of three experiments. RT-PCR analysis revealed that the osteogenic medium significantly induced BMPR-IA and BMPRII expression in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively. *Compared with CM at the same time point, p < 0.05. †Compared with BMP-2 at the same time point, p < 0.05. CM, control medium; OM, osteogenic medium; BMP-2, bone morphogenetic protein-2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; BMPRIA, type IA receptor of BMPR; BMPRII, type II receptor of BMPR; RT-PCR, reverse transcription-polymerase chain reaction; BMSCs, bone marrow stromal cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2908870&req=5

Figure 4: mRNA expression of the BMP receptor in BMSCs. BMSCs were treated with CM or OM with or without BMP-2. mRNA was extracted from cells on day 3, 6, and 9 and analyzed by RT-PCR. GADPH mRNA was used as an internal control to normalize the amount of RNA. The relative expression level of mRNA was depicted as the ratio of the density of mRNA to GADPH mRNA at the same time point. The results correspond to a representative of three experiments. RT-PCR analysis revealed that the osteogenic medium significantly induced BMPR-IA and BMPRII expression in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively. *Compared with CM at the same time point, p < 0.05. †Compared with BMP-2 at the same time point, p < 0.05. CM, control medium; OM, osteogenic medium; BMP-2, bone morphogenetic protein-2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; BMPRIA, type IA receptor of BMPR; BMPRII, type II receptor of BMPR; RT-PCR, reverse transcription-polymerase chain reaction; BMSCs, bone marrow stromal cells.
Mentions: Because BMP signaling has been shown to promote the osteoblastic differentiation of BMSCs, BMP receptors were assessed by RT-PCR during BMSC differentiation. As shown in Fig. 4, the expression levels of BMPR-IA in OM with or without BMP-2 were significantly higher than CM or BMP-2 at three or six days of stimulation (p < 0.05). No difference was found between the CM and BMP-2 groups. Nine days after stimulation, the level of BMPRIA mRNA was continuously induced in OM with or without BMP-2 (p < 0.05). The expression of BMPRIA significantly increased in the BMP-2 groups as compared to the other three groups at nine days of stimulation (p < 0.05).

Bottom Line: Bone morphogenetic protein (BMP) signaling has important effects on the process of skeletogenesis.The OM significantly induced the expression of type IA receptor of BMPR (BMPRIA) and type II receptor of BMPR (BMPRII) in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively.In addition, BMP signaling through BMPRIA and BMPRII regulates the osteogenic differentiation of rat BMSCs in OM with or without BMP-2.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital, Sun Yat-sen University, 58 Zhongshan Road II, Guangzhou, 510080, PR China. anxunwang@yahoo.com

ABSTRACT

Purpose: Several signaling pathways have been shown to regulate the lineage commitment and terminal differentiation of bone marrow stromal cells (BMSCs). Bone morphogenetic protein (BMP) signaling has important effects on the process of skeletogenesis. In the present study, we tested the role of bone morphogenetic protein receptor (BMPR) in the osteogenic differentiation of rat bone marrow stromal cells in osteogenic medium (OM) with or without BMP-2.

Materials and methods: BMSCs were harvested from rats and cultured in OM containing dexamethasone, beta-glycerophosphate, and ascorbic acid, with or without BMP-2 in order to induce osteogenic differentiation. The alkaline phosphatase (ALP) activity assay and von kossa staining were used to assess the osteogenic differentiation of the BMSCs. BMPR mRNA expression was assessed using reverse transcriptionpolymerase chain reaction (RT-PCR).

Results: The BMSCs that underwent osteogenic differentiation in OM showed a higher level of ALP activity and matrix mineralization. BMP-2 alone induced a low level of ALP activity and matrix mineralization in BMSCs, but enhanced the osteogenic differentiation of BMSCs when combined with OM. The OM significantly induced the expression of type IA receptor of BMPR (BMPRIA) and type II receptor of BMPR (BMPRII) in BMSCs after three days of stimulation, while BMP-2 significantly induced BMPRIA and BMPRII in BMSCs after nine or six days of stimulation, respectively.

Conclusion: BMSCs commit to osteoblastic differentiation in OM, which is enhanced by BMP-2. In addition, BMP signaling through BMPRIA and BMPRII regulates the osteogenic differentiation of rat BMSCs in OM with or without BMP-2.

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