• Title, Summary, Keyword: Bone morphogenetic protein-2

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Effects of Cervi Pantotrichum Cornu and Cervi Cornu on Longitudinal Bone Growth in Adolescent Male Rats (녹용(鹿茸)과 녹각(鹿角)의 성장기 흰쥐 장골 길이성장에 대한 효과)

  • Kim, Ki-Tae;Kim, Myung-Gyou;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.21 no.1
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    • pp.63-69
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    • 2006
  • Objectives : This study was designed to investigate the effects of Cervi Pantotrichum Cornu and Cervi Cornu on the growth of longitudinal bone in the adolescent male rats. Methods : Longitudinal bone growth was measured by fluorescence microscopy. To examine the effects on the growth plate metabolism, the heights of growth plate and the induction of local bone morphogenetic protein-2 were measured. Results : Treatment of Cervi Pantotrichum Cornu significantly enhanced longitudinal bone growth compared with control group. However, Cervi Cornu did not show the significant effects. Conclusion : Cervi Pantotrichum Cornu enhanced longitudinal bone growth and promoted the induction of local bone morphogenetic protein-2 of growth plate in adolescence male rats.

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Soluble expression and purification of synthetic human bone morphogenetic protein-2 in Escherichia coli

  • Ihm, Hyo-Jin;Yang, Seung-Ju;Huh, Jae-Wan;Choi, Soo-Young;Cho, Sung-Woo
    • BMB Reports
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    • v.41 no.5
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    • pp.404-407
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    • 2008
  • A 345-bp gene that encodes human bone morphogenetic protein-2 (hBMP-2) has been synthesized. The codon usage of the resulting gene was modified to include those triplets that are utilized in highly expressed Escherichia coli genes. The hBMP-2 gene was efficiently expressed in E. coli as a soluble and active protein. Since the recombinant hBMP-2 was readily solublized, no further solublization steps were required throughout purification. No additional tagging residues were introduced into the synthetic hBMP-2 gene product. The developed synthetic gene is a promising approach for scaling-up the soluble expression of hBMP-2.

Effect of protein transduction domain fused-bone morphogenetic protein-2 on bone regeneration in rat calvarial defects (단백질 전달 영역 융합-Bone Morphogenetic Protein-2가 백서 두개골 결손부에서 골 조직 재생에 미치는 효과)

  • Um, Yoo-Jung;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho;Chai, Jung-Kiu;Kim, Chang-Sung
    • Journal of Periodontal and Implant Science
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    • v.38 no.2
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    • pp.153-162
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    • 2008
  • Purpose: Recombining bone morphogenetic protein (BMP) is usually acquiredfrom high level animals. Though this method is effective, its high cost limits its use. The purpose of this study was to evaluate the effect of bone morphogenetic protein-2 with protein transduction domain (BMP-2/PTD;TATBMP-2) on bone regeneration. Rat calvarial defect model and osteoblastic differentiation model using MC3T3 cell were used for the purpose of the study. Materials and Methods: MC3T3 cells were cultured until they reached a confluence stage. The cells were treated with 0, 0.1, 1, 10, 100, 500 ng/ml of BMP-2/PTD for 21 days and at the end of the treatment, osteoblastic differentiation was evaluated usingvon Kossa staining. An 8mm, calvarial, critical-size osteotomy defect was created in each of 48 male Spraque-Dawley rats (weight $250{\sim}300\;g$). Three groups of 16 animals each received either BMP-2/PTD (0.05mg/ml) in a collagen carrier, collagen only, or negative surgical control. And each group was divided into 2 and 8 weeks healing intervals. The groups were evaluated by histologic analysis(8 animals/group/healing intervals) Result: In osteoblastic differentiation evaluation test, a stimulatory effect of BMP-2/PTD was observed in 10ng/ml of BMP-2/PTD with no observation of dose-dependent manner. The BMP-2/PTD group showed enhanced local bone formation in the rat calvarial defect at 2 weeks. New bone was observed at the defect margin and central area of the defect. However, new bone formation was observed only in 50% of animals used for 2weeks. In addition, there was no new bone formation observed at 8 weeks. Conclusion: The results of the present study indicated that BMP-2/PTD(TATBMP-2) have an positive effect on the bone formation in vitro and in vivo. However, further study should be conducted for the reproducibility of the outcomes.

Expression and Purification of Recombinant Human Bone Morphogenetic Protein-7 (rhBMP-7) in Bacillus subtilis (고초균을 이용한 재조합 인간 골 형성 단백질-7의 발현과 정제)

  • Kim, Chun-Kwang;Oh, Sung-Duk;Rhee, Jong-Il
    • KSBB Journal
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    • v.25 no.3
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    • pp.257-264
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    • 2010
  • Bone morphogenetic protein-7 (BMP-7) is one of important growth factors for skeletal development and bone growth. In this work, BMP-7 was efficiently expressed in recombinant Bacillus subtilis. The mature BMP-7 protein indicated molecular weight of 15.4 kDa by Western blot assay and was secreted into culture medium with 0.35 ng/mL. The extracellular and intracellular rhBMP-7 proteins were purified by using a FPLC system with an ion exchange column and a gel filtration column. The extracellular and intracellular rhBMP-7 proteins had finally a 57.1% purity and a 36.2% purity, respectively. The purified rhBMP-7 proteins showed an intact biological activity which stimulated alkaline phophatase (ALP) activity in MC3T3-E1 cells.

Ulnar Radial Nonunion Fracture Treated with Recombinant Human Bone Morphogenetic Protein-2 in a Dog (개의 요.척골유합부전의 Recombinant Human Bone Morphogenetic Protein-2 적용 치료례)

  • 홍성혁
    • Journal of Veterinary Clinics
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    • v.18 no.2
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    • pp.156-159
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    • 2001
  • A 6-year-old male mongrel dog with a 7-month history of ulnar-radial nonunion fracture was treated with implantation of recombinant human bone morphogenetic protein-2 (rhBMP-2). The dog had received surgical correction three times prior to the admission but radiography of the affected limb revealed a typical figure of nonunion fracture. Glossly, the fractured ends were sclerotic and the area between the ends was filled with fibrous tissue. After debridement the shaft was fixed by an 10-hole plate. rhBMP-2 at a total dose of 256 micrograms was implanted with a synthetic carrier into the 10-mm defect formed by the debridement. Callus formation responding to rhBMP-2 was radiographically observed at 4 weeks after implantation and the defect bridged both fracture ends by 8 weeks after implantation. The plate was removed at 12 months after implantation. Any complications were not observed for 5 months after removal of the plate.

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Bone regenerative effects of recombinant human bone morphogenetic protein-2 employed protein transduction domain (Protein transduction domain을 이용한 recombinant human bone morphogenetic protein-2의 골재생효과)

  • Jung, Sung-Won;Kim, Nam-Hee;Yook, Jong-In;Kim, Chang-Sung;Kim, Hyung-Jun;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
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    • v.37 no.3
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    • pp.497-509
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    • 2007
  • Bone morphogenetic proteins(BMPs) are regarded as members of the transforming growth $factor-{\beta}$ superfamily with characteristic features in their amino acid sequences. A number of studies have demonstrated the biologic activities of BMPs, which include the induction of cartilage and bone formation. Recently there was a attempt to overcome a limitation of mass production, and economical efficieny of rh-BMPs. The method producing PTD by using bacteria have advantages of acquiry a mass of proteins. Hences, a new treatment which deliver protein employed by protein transduction domain(PTD) has been tried. The purpose of this study was to evaluate the bone regenerative effect of TATBMP-2 and TAT-HA2-BMP-2 employed by PTD from HlV-1 TAT protein for protein translocation in the rat calvarial model. An 8mm calvarial, critical size osteotomy defect was created in each of 32 male Spraque-Dawley rats(weight $250{\sim}300g$). The animals were divided into 4 groups of 32 animals each (4 animals/group/healing interval). The defect was treated with TATBMP-2/ACS(Absorbable collagen sponge) (TATBMP-2 0.1mg/ml), TAT-HA2-BMP-2/ACS(TAT-HA2-BMP-2 0.1mg/ml), ACS alone or left untreated for surgical control(negative control). The rats were sacrificed at 2 or 8 weeks postsurgery, and the results were evaluated histologically. The results were as follows: New bone formation were not significantly greater in the TATBMP-2/ACS group relative to negative, and positive control groups. New bone was evident at the defect sites in TAT-HA2-BMP-2/ACS group relative to negative, positive control and TATBMP-2 groups. There were a little bone regeneration in TATBMP-2 groups. While, enhanced local bone formation were observed in TAT-HA2-BMP-2 group. But, The results was not the same in all rat defects. Therefore, further investigations are required to develop a method. which disperse homogenously, and adhere to target cells.

Recombinant Human Bone Morphogenetic Protein-2 in Development and Progression of Oral Squamous Cell Carcinoma

  • Zaid, Khaled Waleed;Chantiri, Mansour;Bassit, Ghassan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.927-932
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    • 2016
  • Bone morphogenetic proteins (BMPs), belonging to the transforming growth factor-${\beta}$ superfamily, regulate many cellular activities including cell migration, differentiation, adhesion, proliferation and apoptosis. Use of recombinant human bone morphogenic protein-2 (rhBMP-2) in oral and maxillofacial surgery has seen a tremendous increase. Due to its role in many cellular pathways, the influence of this protein on carcinogenesis in different organs has been intensively studied over the past decade. BMPs also have been detected to have a role in the development and progression of many tumors, particularly disease-specific bone metastasis. In oral squamous cell carcinoma - the tumor type accounting for more than 90% of head and neck malignancies- aberrations of both BMP expression and associated signaling pathways have a certain relation with the development and progression of the disease by regulating a range of biological functions in the altered cells. In the current review, we discuss the influence of BMPs -especially rhBMP-2- in the development and progression of oral squamous cell carcinoma.

Bone Morphogenetic Protein 2-induced MAPKs Activation Is Independent of the Smad1/5 Activation

  • Jun, Ji-Hae;Ryoo, Hyun-Mo;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.34 no.2
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    • pp.115-121
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    • 2009
  • Bone morphogenetic protein (BMP) 2 is a potent osteogenic factor. Although both Smad1/5 and mitogenactivated protein kinases (MAPKs) are activated by BMP2, the hierarchical relationship between them is unclear. In this study, we examined if BMP2-stimulated MAPK activation is regulated by Smad1/5 or vice versa. When C2C12 cells were treated with BMP2, the activation of extracellular signal-regulated kinase (ERK), p38 MAPK and c-Jun-N-terminal kinase was evident within 5 min. The knockdown of both Smad1 and Smad5 by small interfering RNA did not affect the activation of these MAPKs. In addition, neither the overexpression of Smad1 nor Smad5 induced ERK activation. When ERK activation was induced by constitutively active MEK1 expression, the protein level and activation of Smad1 increased. Furthermore, the inhibition of constitutively active BMP receptor type IB-induced ERK activation significantly suppressed Smad1 activation. These results indicate that Smad1/5 activation is not necessary for BMP2-induced MAPK activation and also that ERK positively regulates Smad1 activation.

Expression of Recombinant Human Bone morphogenetic protein 2 (hBMP2) in Insect cells

  • Kim, Seong-Wan;Kim, Seong-Ryul;Park, Seung Won;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology
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    • v.34 no.1
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    • pp.1-5
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    • 2017
  • Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

Demineralized dentin matrix combined with recombinant human bone morphogenetic protein-2 in rabbit calvarial defects

  • Um, In-Woong;Hwang, Suk-Hyun;Kim, Young-Kyun;Kim, Moon-Young;Jun, Sang-Ho;Ryu, Jae-Jun;Jang, Hyon-Seok
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.42 no.2
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    • pp.90-98
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    • 2016
  • Objectives: The aim of this study was to compare the osteogenic effects of demineralized dentin matrix (DDM) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) in rabbit calvarial defects with DDM and anorganic bovine bone (ABB) combined with rhBMP-2. Materials and Methods: Four round defects with 8-mm diameters were created in each rabbit calvaria. Each defect was treated with one of the following: 1) DDM, 2) ABB/rhBMP-2, or 3) DDM/rhBMP-2. The rhBMP-2 was combined with DDM and ABB according to a stepwise dry and dip lyophilizing protocol. Histological and microcomputed tomography (${\mu}CT$) analyses were performed to measure the amount of bone formation and bone volume after 2- and 8-week healing intervals. Results: Upon histological observation at two weeks, the DDM and ABB/rhBMP-2 groups showed osteoconductive bone formation, while the DDM/rhBMP-2 group showed osteoconductive and osteoinductive bone formation. New bone formation was higher in DDM/rhBMP-2, DDM and ABB decreasing order. The amounts of bone formation were very similar at two weeks; however, at eight weeks, the DDM/rhBMP-2 group showed a twofold greater amount of bone formation compared to the DDM and ABB/rhBMP-2 groups. The ${\mu}CT$ analysis showed markedly increased bone volume in the DDM/rhBMP-2 group at eight weeks compared with that of the DDM group. Notably, there was a slight decrease in bone volume in the ABB/rhBMP-2 group at eight weeks. There were no significant differences among the DDM, ABB/rhBMP-2, and DDM/rhBMP-2 groups at two or eight weeks. Conclusion: Within the limitations of this study, DDM appears to be a suitable carrier for rhBMP-2 in orthotopic sites.