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

OBJECTIVETo investigate the effect of hypoxia on isolated human periodontal ligament stem cells (hPDLSC) in vitro.

METHODShPDLSC were exposed to normoxia (20% O(2)) and hypoxia (1.5% - 2% O(2)). Total cellular RNA and protein were collected on day 1, 2, 4 and 6 after culture. Western analysis and semi-quantitative RT-PCR were used to analyze osteogenic differentiation of hPDLSC, including alkaline phosphatase (ALP), osteocalcin (OCN), secreted protein acidic and rich in cysteine (SPARC) and bone morphogenetic protein-2 (BMP-2).

RESULTSIn the first two days hypoxia slightly increased the growth of hPDLSC [A value was 0.697(hypoxia) vs 0.617 (nomoxia)] and after the third day hypoxia dramatically decreased the growth of the hPDLSC [A value was 0.870(hypoxia) vs 1.242 (nomoxia)]. Up to 90% reduction of ALP activity was observed in hPDLSC after 5 days of hypoxia [A value was 0.004(hypoxia) vs 0.049(nomoxia)]. Hypoxia decreased SPARC expression at protein level and down-regulated ALP, OCN and BMP-2 expression at mRNA level in comparison with nomoxia.

CONCLUSIONSHypoxia inhibited proliferation of hPDLSC and down-regulated BMP-2 mRNA expression, then down-regulated their target genes such as ALP, OCN, and SPARC, thus inhibiting critical steps in osteogenic differentiation of hPDLSC.

Bone Morphogenetic Protein 2 ; metabolism ; Cell Differentiation ; Cell Hypoxia ; Cells, Cultured ; Humans ; Periodontal Ligament ; cytology ; metabolism ; Stem Cells ; cytology ; metabolism

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

OBJECTIVETo investigate the effect of systematic administration of simvastatin on the bone morphogenetic protein-2 (BMP-2) expression in the periodontal tissue after rat tooth movement and on the relapse of tooth movement.

METHODSOrthodontic tooth movement of upper first molar was performed in 32 rats with coil spring for 21 days. The 32 rats were randomly allocated into 4 groups: negative control group (isotonic saline) and three experimental groups (2.5 mg x kg(-1), 5.0 mg x kg(-1) and 10.0 mg x kg(-1)). The simvastatin started to be administered to the experimental groups 1 day before appliances were removed, and once a day there after for 4 weeks. The negative control group received the isotonic saline only. The interdental distance between the first and second maxillary molars were measured, when appliances were removed, and 1 week and 4 weeks after that. After the rats were sacrificed, sections of first maxillary molar and periodontal tissue were studied by immunohistochemistry.

RESULTSThe number and percentage of relapse was lower in the three experimental groups than in the negative control group (P < 0.05, P < 0.01). The lower dose was given, the less relapse there was, with the lowerest dose resulting in lowest percentage of relapse (26.81% and 53.38%). BMP-2 expression in experimental groups was higher than in the negative control group, with the lowerest dose group showing the highest expression (P < 0.001). The BMP-2 expression on the tension side was slightly stronger than that on the compression side (P > 0.05).

CONCLUSIONSSystemic administration of simvastatin could decrease the extent of relapse of the orthodontic-moved tooth in rat, and the lower-dose of simvastatin seemed more effective. The possible mechanism for this may be that simvastatin functions by increasing the expression of BMP-2 in the periodontal tissue, accelerating the osteoblast activity and promoting bone formation.

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Male ; Periodontium ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Simvastatin ; pharmacology ; Tooth Movement Techniques

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