OBJECTIVEThis research explores the regulatory role of bone morphogenetic protein 2 (BMP2) in the expression of sclerostin in OCCM-30 cementoblast.

METHODSOCCM-30 cementoblasts were treated with 50 and 100 ng · mL⁻¹ BMP2 for 3, 5, and 7 days. SOST mRNA was detected by real-time quantitative polymerase chain reaction (RT-PCR). Western blot analysis was employed to detect the sclerostin levels in the nucleus. Five groups were prepared for the experiments: control, BMP2, BMP2+dorsomorphin, BMP2+SB202190, and BMP2+PD98059. OCCM-30 was pretreated with BMP2 for 3 and 5 days, and then the sclerostin and SOST mRNA levels were measured.

RESULTSRT-PCR and Western blot analyses showed that BMP2 upregulated the expression of SOST in a concentration-dependent manner. SOST expression increased with time (P < 0.05). Moreover, sclerostin levels of BMP2+dorsomorphin, BMP2+SB202190, and BMP2+PD98059 groups were lower than that of the BMP2 group, and the sclerostin level in BMP2+dorsomorphin group was lowest (P < 0.05).

CONCLUSIONThe upregulation of SOST by BMP2 in OCCM-30 is mainly mediated by the BMP2/Smad signal pathway.

Blotting, Western ; Bone Morphogenetic Protein 2 ; metabolism ; Bone Morphogenetic Proteins ; metabolism ; Dental Cementum ; metabolism ; Genetic Markers ; Signal Transduction ; Up-Regulation

OBJECTIVETo explore reciprocal action between BMP-2 (bone morphogenetic protein-2) and BMP-3 for better understanding of the mechanism of BMP during bone fracture union.

METHODSrhBMP-2 was added into the cultured fibroblasts with the concentration of 1,200 ng/ml. The expression of BMP-3 in fibroblasts was detected by immunohistochemistry. Eukaryotic expression vector pcDNA3-BMP-3 was transfected into the fibroblasts. After the effective expression of BMP-3 was identified, BMP-2 was also detected by immunohistochemistry in BMP-3 expression cells. The fibroblasts transfected with empty vector pcDNA3 were used as the control.

RESULTSExogenous rhBMP-2 could promote the expression of BMP-3 in fibroblasts. BMP-3 also could be detected in these cells.

CONCLUSIONSBMP-2 and BMP-3 could reciprocally adjust the expression in fibroblasts.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein 3 ; Bone Morphogenetic Proteins ; metabolism ; Cells, Cultured ; Fibroblasts ; metabolism ; Fracture Healing ; physiology ; Immunohistochemistry ; Osteogenesis ; physiology ; Transforming Growth Factor beta

BACKGROUNDThe palate is differently regulated and developed along the anterior-posterior axis. The Bmp signal pathway plays a crucial role in palatogenesis. Conditioned-inactivation of Bmp type I receptor Alk2 or Alk3 in the neural crest or craniofacial region leads to palatal cleft in mice. However, how different Bmp members are involved in palatogenesis remains to be elucidated. In the present study, mRNA expression patterns of Bmp2, Bmp3 and Bmp4 in the developing anterior and posterior palates were examined and compared, focusing on the fusion stage.

METHODSTo detect the expression of Bmp mRNA, antisense riboprobes were synthesized by in vitro transcription. Radioactive in situ hybridization was performed on sagital and coronal sections of mice head from E13 to E18.

RESULTSThe expression of these Bmps were developmentally regulated in the anterior and posterior palates prior to, during and after palatal fusion. During palatal fusion, Bmp4 expression shifted from the anterior to the posterior palate, Bmp2 was highly expressed in both the anterior and posterior palates in this process, whereas Bmp3 was only localized in the posterior palate. They showed generally non-overlapping pattern in their expression domains. Thereafter, their expression was detected in both the anterior and posterior palates regulating osteogenesis and myogenesis respectively.

CONCLUSIONSBmp signalling is involved in palatogenesis in multiple stages and has multiple roles in regulating anterior and posterior palatal development. Disturbances of Bmp signalling during palatogenesis might be a possible mechanism of cleft palate.

Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein 3 ; Bone Morphogenetic Protein 4 ; Bone Morphogenetic Proteins ; genetics ; Female ; Gene Expression Regulation, Developmental ; Mice ; Palate ; embryology ; metabolism ; RNA, Messenger ; analysis ; Signal Transduction ; Transforming Growth Factor beta ; genetics

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Bone Morphogenetic Protein 4 ; metabolism ; Carrier Proteins ; pharmacology ; Liver ; metabolism ; Liver Regeneration ; Male ; Rats ; Rats, Wistar

OBJECTIVETo determine the expression of bone morphogenetic protein 2 and 4 (BMP-2 and BMP-4) in prostatic carcinoma (PCa) and investigate their relationship with clinical stage and Gleason score of tumor.

METHODSForty-eight PCa cases and 5 normal prostatic tissue were analysed for the expressions of BMP-2 and BMP-4 by Western bolt assay.

RESULTSThe optical densities of BMP-2 expressions in the tumor with Gleason score < or =5, 6-8, and > or = 9 were 7547.1 +/- 1964.12, 9657.4 +/- 2010.54, 12467.7 +/- 2496.75 and of BMP-4 expressions were 5174.4 +/- 1400.54, 5940.3 +/- 1587.42, 6332.1 +/- 1647.83, respectively. The optical densities of BMP-2 expressions in the tumor in T1 - T2 and T3 - T4 stages were 8003.37 +/- 1889.23, 12385.55 +/- 2506.72 and of BMP4 expressions were 5267.41 +/- 1 464.19, 6543.75 +/- 1668.46, respectively. There were significant differences between tissues with Gleason score < or =5 and > or =9 (P <0.01), and tissues in T1 - T2 and T3 - T4 stages, in expressions of BMP-2 protein. The expression of BMP-2 protein was significantly high in the PCa with bone metastasis compared with that without bone metastasis.

CONCLUSIONThe expressions of BMP-2 and BMP-4 increase with the progression of clinical stage and Gleason score compared with normal prostatic tissue. The expression of BMP-2 protein is significantly upregulated in bone metastasis of PCa, which indicates a poor prognosis.

Aged ; Aged, 80 and over ; Blotting, Western ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein 4 ; Bone Morphogenetic Proteins ; biosynthesis ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prostate ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Transforming Growth Factor beta ; biosynthesis

OBJECTIVE: To investigate the expressions of transforming growth factor beta1 (TGF-beta1) and bone morphogenetic protein-2 (BMP-2) in human mandible fracture callus and their quantity changes in the process of healing. METHOD: Thirty callus samples from the fractured mandible bone stumps were collected during operation, and two callus samples were collected from the angle-square jaw patients as controls. The expressions of TGF-beta1 and BMP-2 were test by the immunohistochemistry technic-SABC-staining in different periods of human fractured mandibular callus and in osseous tissue of normal angle of mandible. RESULT: The TGF-beta1 and BMP-2 were expressed in callus of different periods but not in normal bone tissue. The expression of TGF-beta1 increased slowly during the first three weeks after fracture and reached its maximum in the third week, and then weakened gradually. The expression of BMP-2 increased gradually during the first two weeks after fracture and reached its maximum in the second week, then the expression weakened gradually. CONCLUSION (1) BMP-2 may be one of the factors promoting the repair of fracture. (2) TGF-beta1 could be another signal pathway in repairment of fracture. (3) There could exist some synergistic effects between TGF-beta1 and BMP-2 in the process of fracture healing.
Adult ; Bone Morphogenetic Protein 2 ; metabolism ; Fracture Healing ; Humans ; Mandibular Fractures ; metabolism ; Middle Aged ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo characterize the role of Smads proteins in alpha 2 (I) collagen (COL1A2) gene expression induced by bone morphogenetic protein-2 (BMP-2) in odontoblast cell line MDPC-23.

METHODSEndogenous Smad protein expression was determined by immunocytochemistry. Smads function and their role in COL1A2 gene expression were investigated in cotransfection experiments using promoter-luciferase reporter gene construct.

RESULTSMDPC-23 cells expressed Smad1, Smad5 and Smad6. BMP-2 promoted the activation of COL1A2 promoter reporter construct. Transient overexpression of Smad1 or Smad5 was enhanced, while overexpression of Smad6 inhibited BMP-2-induced COL1A2 promoter activity. BMP-2 inducibility could be blocked by overexpression of Smad1 or Smad5 dominant negative mutant.

CONCLUSIONSSmad signaling is functioning and appears to be involved in BMP-2-induced COL1A2 collagen transcription in MDPC-23. Smad signaling may play an important role in odontoblast differentiation and dentin extracellular matrix formation mediated by BMP-2.

Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; genetics ; Cell Line ; Collagen ; genetics ; Collagen Type I ; Mice ; Odontoblasts ; cytology ; metabolism ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; genetics

The purpose of this study was to identify specific microRNAs (miRNAs) during differentiation and maturation of interneurons and to predict their possible functions by analyzing the expression of miRNAs during in vitro differentiation of the rat interneuron precursor cell line GE6. In the experiment, the interneuron precursor cell line GE6 was cultured under three different conditions, i. e. the first was that had not added growth factors and the normal differentiation cultured for 4 days (Ge6_4d); the second was that cultured with bone morphogenetic protein-2 (BMP2) for 4 days (Ge6_bmp2); and the third was that cultured with sonic hedgehog (SHH) for 4 days (Ge6_ shh). In addition, another group of undifferentiated GE6 (Ge6_u) was applied as a control. We found in this study that the expression levels of a large number of miRNAs changed significantly during GE6 differentiation. The expression levels of miR-710, miR-290-5p and miR-3473 increased in the GE6 cells with secreted factor BMP2. These miRNAs may play important regulatory roles during interneuron differentiation.
Animals ; Bone Morphogenetic Protein 2 ; chemistry ; Cell Differentiation ; Cell Line ; Hedgehog Proteins ; chemistry ; Interneurons ; cytology ; metabolism ; MicroRNAs ; metabolism ; Rats

OBJECTIVE: To explore the difference of bone formation potential between hypertrophic nonunion tissue and atrophic nonunion tissue, which may be beneficial to nonunion therapy. METHODS: From October 2010 to March 2014, 40 nonunion tissue samples were collected in Department of Orthopedics, Second Xiangya Hospital. The samples were divided into a hypertrophic nonunion group (n=20) and an atrophic nonunion group (n=20) according to nonunion character; or a 20 to 35 years old group (n=18), a 36 to 50 years old group (n=18), a more than 50 years old group (n=4) according to different ages; or a 9-12 months group (n=21), 13-24 months group (n=14) and a more than 24 months group (n=5) according to different nonunion time. Semi-quantification was performed by SP immunohistochemical method and IPP6.0 was used to analyze the expression of bone morphogenetic protein-2 (BMP-2) through measuring the mean optical density. RESULTS: The mean optical density of BMP-2 was 0.1540±0.0408 in hypertrophic nonunion tissue, 0.1372±0.0372 in atrophic nonunion tissue, there was no significant difference between the 2 groups (P>0.05). The mean optical density of BMP-2 was 0.1477±0.0379 in the 20 to 35 years old group, 0.1419±0.0399 in the 35 to 50 years old group, 0.1456±0.0595 in the more than 50 years old group, there was no significant difference among the three groups (P>0.05). The mean optical density of BMP-2 was 0.1449±0.0366 in the 9-12 months group, 0.1472±0.0400 in the 13-24 months group, 0.1445±0.0541 in the more than 24 months group, there was no significant difference among the 3 groups (P>0.05). CONCLUSION The present results suggest that the hypertrophic nonunion tissue share similar osteogenic potential with the atrophic nonunion tissue.
Adult ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Fracture Healing ; Fractures, Ununited ; genetics ; metabolism ; Humans ; Middle Aged ; Osteogenesis ; Young Adult

OBJECTIVETo observe the expression of asporin, bone morphogenetic protein-2 (BMP-2) and alkaline phosphatase (ALP) in cultured human periodontal ligament cells in vitro under relative centrifugal force.

METHODSHuman periodontal ligament cell was cultured in vitro and applied 30 ×g centrifugal force for 0, 1, 2, 6, 10 hours. The expression of asporin, BMP-2 and ALP was observed with real time-PCR.

RESULTSAll the cells showed normal figuration. The expression of asporin, BMP-2, ALP in no force loading group did not have any statistical significance change (P > 0.05). In 1 hour force loading group, the expressions of asporin and BMP-2 were 0.50 ± 0.05 and 0.40 ± 0.13. In 2 hour force loading group, the expressions of asporin and BMP-2 were 0.42 ± 0.09 and 0.58 ± 0.19, which decreased significantly from no force loading group (P < 0.05). The expression of asporin and BMP-2 increased significantly in 6 hour force loading group than in 1 and 2 hour force loading groups (P < 0.05). Then the expression of asporin decreased to no force loading group level (P > 0.05) and the expression of BMP-2 decreased rapidly lower than no force loading group level (P < 0.05). During the 10 hour interval of stress loading, the expression of asporin and BMP-2 showed a positive correlation (r = 0.995, P < 0.05).

CONCLUSIONSThe expression of asporin in human periodontal ligament cell was stable. Short and light centrifugal force could up-regulate the expression of asporin rapidly, and suppress the abnormal BMP-2 expression back to baseline level.

Alkaline Phosphatase ; biosynthesis ; Bone Morphogenetic Protein 2 ; biosynthesis ; Cell Line ; Centrifugation ; Gene Expression ; Humans ; Periodontal Ligament ; metabolism ; Up-Regulation