Bone morphogenetic protein (BMP) homodimers are of significant osteoinductivity. However, their clinical application is limited because of high effective dosage. Recently, BMP heterodimers are reported to address the issue. This is a review of the researches on BMP heterodimers, including existent evidences, types and synthetic methods, biological activities in comparison to BMP homodimers and possible mechanisms, further research direction and future expectations.
Animals ; Biopolymers ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein 7 ; Bone Morphogenetic Proteins ; genetics ; pharmacology ; Humans ; Protein Multimerization

OBJECTIVETo observe the effect of the self-made chitosan thermosensitive hydrogel system with dual-release bone morphogenetic protein and chlorhexidine on periodontal defects repair.

METHODSThe furcation defect model was established on dog premolar. The models were divided into five groups, including three experimental groups, one control group and one blank control group. The hydrogel with the chlorhexidine/3-cyclodextrin inclusion complexes (IC) /rhBMP-2, hydrogel with rhBMP-2, hydrogel with IC, the pure hydrogel were applied to the defects of the four groups, respectively, and the blank control group did not receive any agent. The dogs were sacrificed 8 weeks later and the periodontal regeneration and gingival condition were observed by histological examination.

RESULTSObvious periodontal tissue regeneration was found in group one and two. The heights of new bone reached 99.2% of the defects in group one, 87.8%, 63.6%, 37.0% and 34.3% in group two, three, four and blank control groups, respectively. The inflammation of the affected gingiva showed less significant in group one and group three than in the other groups.

CONCLUSIONSrhBMP-2 and chlorhexidine played their independent role in repairing periodontal defects and the dual-release chitosan thermosensitive hydrogel system is effective and convenient to use.

Animals ; Bone Morphogenetic Proteins ; pharmacology ; Chitosan ; pharmacology ; Chlorhexidine ; pharmacology ; Dogs ; Hydrogels ; pharmacology ; Male ; Periodontium ; drug effects ; physiology ; Regeneration ; Tissue Engineering

Optimization of cultivation condition of recombinant E. coli DH5 alpha/pDH-B2m and the condition suitable for expression of recombinant mature peptide of human bone morphogenetic protein-2 was carried out in 500 mL shaking flasks and then transferred to NBS Bioflo IV, a 20 L DO feed-back fed-batch culture system, to obtain rhBMP-2. The results indicate that keeping dissolved oxygen at 40% and controlling nutrient feeding rate with DO feed back strategy can obtain theoretically 3.59 g recombinant mature peptide of hBMP-2 per liter of broth, the final cell density OD600 reaches 57(22.8 g dry cell weight/L), and the expression of rhBMP-2 amounts to 30% of the total protein in E. coli.
Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; biosynthesis ; genetics ; Culture Media ; Escherichia coli ; genetics ; Fermentation ; Glycerol ; pharmacology ; Recombinant Proteins ; biosynthesis ; Time Factors ; Transforming Growth Factor beta

OBJECTIVE: To investigate the effect of human bone morphogenetic protein (hBMPs) 2/3/6 and 12 on osteosarcoma cell UMR106. METHODS: Adenovirus-BMP2/3/6 and 12 (AdBMP2/3/6 and12) were used to treat the cell line. Their proliferation, apoptosis, and transmigration were detected by Trypan blue exclusion test, TdT-mediated biotinylated-dUTP nick end labeling (TUNEL), acridine orange-ethidium bromide (AO/EB) double fluorescent dye staining, and transwell-room test, respectively. The alkaline phosphatase (ALP) activity was detected to reflect the differentiation of tumors. RESULTS: Compared with the control groups, the cell survival rate of the experimental groups treated with AdBMP2/3/6 and 12 showed a significant time-dependent decrease (P<0.01). The apoptosis indexes were increased significantly (P<0.01) and the results from TUNEL and AO/EB method were consistent. The cell numbers of transmembrane significantly decreased at 24,48, and 72 h (P<0.01). AdBMP2/3/6 and 12 treatment enhanced the activity of ALP activity from day 3 and this effect might still be observed up to day 9 of the treatment (P<0.01). CONCLUSION hBMPs2/3/6 and 12 can inhibit the proliferation and transmigration, and induce their apoptosis and differentiation in osteosarcoma cell line UMR106.
Adenoviridae ; genetics ; metabolism ; Apoptosis ; drug effects ; Bone Morphogenetic Protein 2 ; pharmacology ; Bone Morphogenetic Protein 3 ; pharmacology ; Bone Morphogenetic Protein 6 ; pharmacology ; Bone Morphogenetic Proteins ; pharmacology ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; drug effects ; Growth Differentiation Factors ; pharmacology ; Humans ; Osteosarcoma ; pathology ; Recombinant Proteins ; pharmacology

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

OBJECTIVEThis study was designed to investigate the optimal combination of known osteogenic biomaterials with shape conforming struts to achieve calvarial vault reconstruction, using a canine model.

METHODSEighteen adolescent beagles were divided equally into 6 groups. A critical size defect of 6 x 2 cm traversed the sagittal suture. The biomaterials used for calvarial reconstruction were demineralised perforated bone matrix (DBM), recombinant human bone morphogenetic protein-2 (rhBMP2) and autogenous platelet-rich plasma (PRP). The struts used were cobalt chrome (metal) or resorbable plate. The groupings were as follows: 1) DBM + metal, 2) DBM + PRP + metal, 3) DBM + PRP + resorbable plate, 4) DBM + rhBMP2 + metal, 5) DBM + rhBMP2 + PRP + metal, and 6) DBM + rhBMP2 + resorbable plate. Animals were euthanised at 3 months post-surgery. There was no mortality or major complications. Analysis was performed macroscopically, histologically, and with computed tomography (CT).

RESULTSThere was complete bony regeneration in the rhBMP2 groups only. Non-rhBMP2 groups had minimal bony ingrowth from the defect edges and on the dural surface, a finding confirmed by CT scan and histology. PRP did not enhance bone regeneration. Shape conformation was good with both metal and resorbable plate.

CONCLUSIONrhBMP2 but not PRP accelerated calvarial regeneration in 3 months. The DBM in the rhBMP2 groups were substituted by new trabecular bone. Shape molding was good with both metal and resorbable plate.

Animals ; Biocompatible Materials ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Bone Regeneration ; physiology ; Dogs ; Models, Animal ; Postoperative Care ; Recombinant Proteins ; pharmacology ; Reconstructive Surgical Procedures ; Skull ; growth & development ; pathology ; surgery ; Transforming Growth Factor beta ; pharmacology

BACKGROUNDThe recombinant human bone morphogenetic protein 2 (rhBMP-2) has been used to induce osteogenesis in animals' middle ear and this technique is possible to be used to reconstruct the defects of ossicles. The side effects of the rhBMP-2 in middle ear should be observed before using in clinic. Thus we prepared the controlled release rhBMP-2 and implanted it into the acoustic bulla of guinea pigs. The effect on the cochlea was observed.

METHODSWe prepared the acellular cancellous bone, accompanied with rhBMP-2. The material accompanied with rhBMP-2 was implanted into one acoustic bulla of the animal and the opposite side of the acoustic bulla was implanted with acellular cancellous bone without rhBMP-2. Totally 20 guinea pigs were undergone this procedure. After the operation, the auditory brainstem response (ABR) of the animals was tested according to the time sequence. Three months after the operation, the animals were sacrificed. The osteogenesis induced by rhBMP-2, the acoustic bulla and cochlea affected by rhBMP-2 were observed. The structures of hair cells were observed after silver nitrate staining.

RESULTSThe animals were recovered soon after surgery. The hearing thresholds of the animals were declined slightly just after the surgery and come back completely after 3 months. Also, the bulla and cochlea were normal in shape. The osteogenesis occurred in the pore of the acellular cancellous bone with rhBMP-2. There was not any abnormal hyperplasia of bone in the bulla and cochlea. The articulation between the stapes and oval window was not merged. The shapes of the hair cells were normal and there was no obvious deletion of the hair cells compared with control group.

CONCLUSIONSThe controlled release rhBMP-2 transplanted into the middle ear could induce osteogenesis in the bulla of the animals. It did not affect the shape of the bulla and the hearing threshold of the animal, and did not induce the abnormal hyperplasia of bone in the bulla and might be used to reconstruct the defects of ossicles.

Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Bone Regeneration ; drug effects ; Cochlea ; physiopathology ; surgery ; Cochlear Implants ; Ear Ossicles ; metabolism ; Guinea Pigs ; Humans ; Osteogenesis ; drug effects ; Recombinant Proteins ; pharmacology ; Transforming Growth Factor beta ; pharmacology

OBJECTIVETo examine the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on NIH3T3 cells.

METHODSThe NIH3T3 cells were cultured in DMEM with 50 microg/L rhBMP-2. The proliferation ability, alkaline phosphatase (ALP) activity and osteocalcin (OC) quantity of NIH3T3 cells were investigated.

RESULTSThe cells showed decreased proliferation ability but enhanced ALP activity and OC quantity.

CONCLUSION50 microg/L rhBMP-2 can transform NIH3T3 fibroblasts to osteo-like cells.

Alkaline Phosphatase ; drug effects ; metabolism ; Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Cell Proliferation ; drug effects ; DNA ; drug effects ; Fibroblasts ; cytology ; Mice ; NIH 3T3 Cells ; Osteocalcin ; drug effects ; Recombinant Proteins ; pharmacology ; Transforming Growth Factor beta ; pharmacology

To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng/mL). After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen II mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen II mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesenchymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
Bone Marrow Cells/*cytology ; Bone Morphogenetic Proteins/*pharmacology ; Cell Differentiation/*drug effects ; Cells, Cultured ; Chondrocytes/*cytology ; Chondrogenesis/drug effects ; Chondrogenesis/physiology ; Mesenchymal Stem Cells/*cytology ; Recombinant Proteins/pharmacology

OBJECTIVETo study the feasibility of osteogenic phenotype expression by human skin fibroblasts induced in polyglycolic acid (PGA) foams and the effect of tumor necrosis factor-alpha (TNF-alpha) on the expression of bone morphogenetic protein (BMP) receptors.

METHODSThe fibroblasts were isolated, purified from human skin. (1) Fibroblasts were seeded onto PGA foams. The cell-PGA complexes were cultured in RCCS for 6 weeks, in the media of TNF-alpha (50 U/ml) and BMP-2 (0.1 microg/ml). 1 d, 3 and 6 weeks later, cells and extracellular matrix were investigated by electron microscopic and histochemistry observation respectively. Secretion of osteogenic markers were analyzed by biochemical methods. (2) Fibroblasts were seeded on the glass fragments or culture flasks and treated with TNF-alpha (50 U/ml) in different usage (one-time, all-time). The RT-PCR method and the immunohistochemistry fluorescence staining were used to examine the influence of TNF-alpha on the mRNA expression and the protein expression of the type I BMP receptors at 2, 4, 6, 8 d after treatment.

RESULTSFibroblasts seeded on the PGA foams formed 3-dimensional matrix 3 weeks after seeding, which was demonstrated as osteo-tissue by tetracycline labeling and ARS staining. Cells secreted much more bone-specific alkaline phosphatase (B-AKP) and osteocalcin (OCN) into supernatant than the cells that were cultured in the media without TNF-a and BMP2. Eight days after all-time usage, the TNF-alpha (50 U/ml) increased the expression of the mRNA and protein of the type IB BMP receptor.

CONCLUSIONSFibroblasts on 3-D cell-foam structures can express osteoblastic phenotype under certain inducing conditions. The numerous fibroblasts in body would be a promising resource for cell seeds candidate of tissue- engineered bone. TNF-alpha provides the essential condition for BMP2's target effect on fibroblasts, and combined use of TNF-alpha and BMP2 is one of the regulating factors.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein Receptors, Type I ; biosynthesis ; genetics ; Bone Morphogenetic Protein Receptors, Type II ; biosynthesis ; genetics ; Bone Morphogenetic Proteins ; pharmacology ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Cells, Cultured ; Drug Synergism ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Phenotype ; Polyglycolic Acid ; Transforming Growth Factor beta ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology