• 제목/요약/키워드: Bone Morphogenetic Protein 4

검색결과 125건 처리시간 0.019초

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

  • 김춘광;오성덕;이종일
    • KSBB Journal
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    • 제25권3호
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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.

치아발육시 방사선조사와 칼슘결핍이 골형성단백질-2/4의 분포에 미치는 영향에 관한 여구 (The Effects of Irradiation and Calcium-deficient Diet on the Expression of Bone Morphogenetic Protein-2/4 during Early Tooth Development)

  • 박대희;황의환;이상래
    • Imaging Science in Dentistry
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    • 제30권3호
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    • pp.169-181
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    • 2000
  • Purpose: To investigate the expression of bone morphogenetic protein (BMP)-2/4 during eary tooth development after irradiation and calcium-deficient diet. Materials and Methods: The pregnant three-week-old Sprague-Dawley rats were used for the study. The control group was non-irradiation/normal diet group (Group 1), and the experimental groups were irradiation/normal diet group (Group 2) and irradiation/calcium-deficient diet group (Group 3). The abdomen of the rats at the 9th day of pregnancy were irradiated with single dose of 350 cGy. The rat pups were sacrificed at embryonic 18 days, 3 days and 14 days after delivery and the maxillae tooth germs were taken. The tissue sections of specimen were stained immunohisto-chemically with anti-BMP-2/4 antibody. Results: At embryo-18 days, immunoreacivity for BMP-2/4 of the Group 1 was modetate in stratum intermedium of dental organ and weak in dental papilla and dental follicle, but that of Group 2 was weak in cell layer of dental organ, and no immunoreacivity was shown in dental papilla and dental follice of Group 2 and in all tissue components of the Group 3. At postnatal-3 days, immunoreacivity for BMP-2/4 of the Group 1 was strong in cell layer of dental organ, odontoblasts and developing alveolar bone, but that of Group of 2 and Group 3 was weak in odontoblasts and developing alveolar bone. At postnatal-14 days, immunoreacivity for BMP-2/4 of the Group 1 was strong in newly formed cementum, alveolar bone and odontoblasts, but that of Group 2 was weaker than that of Group 1. In the Group 3, tooth forming cell layer showed weak immunoreactivity, but other cell layers showed no immunoreactivity. Couclusion : The expression of bone morphogenetic protein (BMP)-2/4 during early tooth development was disturbed after irradiation and calcium-deficient diet.

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

  • 홍성혁
    • 한국임상수의학회지
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    • 제18권2호
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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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Clinical Outcome of Implants Placed in Grafted Maxillary Sinus Using Recombinant Human Bone Morphogenetic Protein-2: A 5-year Follow-Up Study

  • Yu-Jeong Baek;Jin-Ho Lee;Hyo-Jeong Kim;Bok-Joo Kim;Jang-Ho Son
    • Journal of Korean Dental Science
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    • 제17권1호
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    • pp.45-52
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    • 2024
  • Purpose: To investigate the 5-year outcome of dental implants placed in a grafted maxillary sinus using recombinant human bone morphogenetic protein-2 (rhBMP-2). Materials and Methods: We retrospectively analyzed 27 implants after maxillary sinus floor augmentation (MSFA) using rhBMP-2 in 16 patients between January 2016 and March 2017. The study evaluated two outcome variables: (1) 5-year cumulative survival and success rate of the implant after functional loading and (2) marginal bone loss (MBL) for implant failure. Results: The average residual bone height was 4.78±1.53 mm. The healing period before loading was 8.35±2.34 months. The crown-to-implant ratio was 1.31±0.26. The 5-year cumulative survival and success rate after functional loading were 100% and 96.3%, respectively. The 5-year average MLB was 0.89±0.82 mm. Conclusion: Placing dental implants with MSFA using rhBMP-2 is a reliable procedure with favorable long-term survival and success rates.

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

  • 정성원;김남희;육종인;김창성;김형준;조규성
    • Journal of Periodontal and Implant Science
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    • 제37권3호
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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.

성견에서 임프란트 매식시 골형성단백 사용에 따른 골재생에 관한 연구 (A STUDY OF EFFECTS OF BONE MORPHOGENETIC PROTEIN BONE REGENERATION OF IMPLANTS IN DOGS)

  • 조진희;방몽숙;이종호
    • 대한치과보철학회지
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    • 제32권4호
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    • pp.593-607
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    • 1994
  • The purpose of this study is to evaluate the effect of the bone morphogenetic protein, bone matrix gelatin and collagen matrix on the amount and shape of generating new bone adjacent to the implant. Implants were inserted in the mandible of adult dogs at 2 months after teeth extraction. Artificial bony defects, 3mm in width and 4mm in depth were made at the mesial and distal side of implant. Experimental groups were divided into three groups ; Group 1 : Defects filled with collagen matrix and bone morphogenetic protein, Group 2 : Defects filled with bone matrix gelatin. Control group : Defects filled with only collagen matrix. After implantation, the animals were sacrificed at 1,3,5 and 10 weeks for light microscopic examination. For the fluorescent microscopic examination. each tertracycline Hcl and calcein were injected at 1, 3, 5, 8 and 10 weeks after implantation. The results obtained were as follows : 1. The molecular weight of bovine BMP was about 18,100 by hydroxyapatite chromatography. 2. Osseointegration was observed in experimental groups 1 & 2, and BMG and BMP had an excellent bone forming capability as a filling materials to the repair of the bone defects. 3. The degree of healing of bone defect area, the experimental group 1 showed more prominent bone formation than control group, and the control group showed fibrous connective tissue between the implant and the bone. 4. In the fluorescent microscopic findings, bone remodelling was observed regenerative lamellar bone at defect area in experimental group 1, and partial remodelling in experimental group 2, In the control group, fibrous connective tissue was observed between the implant and bone surface and sign of remodelling was not apperaed. Above results suggest that BMP has rapid osteoinductive property and can be used clinically as a bone substitute on bone defects around implants.

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Combined effect of bisphosphonate and recombinant human bone morphogenetic protein 2 on bone healing of rat calvarial defects

  • Kim, Ho-Chul;Song, Jae-Min;Kim, Chang-Joo;Yoon, Sang-Yong;Kim, In-Ryoung;Park, Bong-Soo;Shin, Sang-Hun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제37권
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    • pp.16.1-16.7
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    • 2015
  • Background: This study aimed to investigate new bone formation using recombinant human bone morphogenetic protein 2 (rhBMP-2) and locally applied bisphosphonate in rat calvarial defects. Methods: Thirty-six rats were studied. Two circular 5 mm diameter bony defect were formed in the calvaria using a trephine bur. The bony defect were grafted with $Bio-Oss^{(R)}$ only (group 1, n = 9), $Bio-Oss^{(R)}$ wetted with rhBMP-2 (group 2, n = 9), $Bio-Oss^{(R)}$ wetted with rhBMP-2 and 1 mM alendronate (group 3, n = 9) and $Bio-Oss^{(R)}$ wetted with rhBMP-2 and 10 mM alendronate (group 4, n = 9). In each group, three animals were euthanized at 2, 4 and 8 weeks after surgery, respectively. The specimens were then analyzed by histology, histomorphometry and immunohistochemistry analysis. Results: There were significant decrease of bone formation area (p < 0.05) between group 4 and group 2, 3. Group 3 showed increase of new bone formation compared to group 2. In immunohistochemistry, collagen type I and osteoprotegerin (OPG) didn't show any difference. However, receptor activator of nuclear factor ${\kappa}B$ ligand (RANKL) decreased with time dependent except group 4. Conclusion: Low concentration bisphosphonate and rhBMP-2 have synergic effect on bone regeneration and this is result from the decreased activity of RANKL of osteoblast.

육미지황탕가미방이 골형성 관련 유전자인 TG2와 BMP4의 전사활성에 미치는 영향 (Effect of Yukmigihwang-tang kamibang on the Expression of Osteo-related Genes, TG2 and BMP4)

  • 신용욱;박용일;김홍렬;이응세
    • 대한한의학회지
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    • 제23권2호
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    • pp.190-197
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    • 2002
  • Objectives : This study was performed to examine the effect of Yukmigiwhang-tang kamibang, a mixture of oriental herbal extracts, on the transcription of bone fonnation genes, BMP4 (bone morphogenetic protein 4) and TG2 (transglutaminase-2). Methods : Bone-related cells, MG-63 (human male osteosarcoma), HOS-TE85 (human female osteosarcoma), and KG-l (bone marrow) were cultured with portions of Yukmigiwhang-tang kamibang and the transcription activities of bone-related genes, BMP4 (bone morphogenetic protein 4) and TG2 (transglutaminase-2), were determined by Reverse-Transcription Polymerase Chain Reaction (RT-PCR). Results : Transcription of BMP4 gene in HOS-TE85 cell increased up to 40% at 0.3% (v/v) of Yukmigiwhang- tang kamibang extract and that of TG2 gene in MG-63 cells also increased up to 40% at 0.3-0.4% of the same extract. Although it was less significant when compared to those in other cells, the transcription of BMP4 gene in KG-l cells also increased up to 10 to 25%. Conclusions : These results clearly demonstrated that Yukmigiwhang-tang kamibang have an effect on transcription activity of bone-related genes, TG2 and BMP4, suggesting that it may play an important role in bone formation.

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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 and Biomaterials
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    • 제34권1호
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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.

골형태형성단백질이 백서치주인대세포와 두개관세포에 미치는 영향 (A Study of the Effects of Bone Morphogenetic Protein on the Characteristics of Rat Periodontal Ligament and Calvaria Cells)

  • 최진근;이만섭;권영혁;허익
    • Journal of Periodontal and Implant Science
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    • 제29권4호
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    • pp.765-785
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    • 1999
  • Bone morphogenetic protein-2/4 (BMP-2/4) are members of Transforming Growth $Factor-{\beta}\;(TGF-{\beta})$ superfamily and they may differentiate the osteoprogenitor cell and induce formation of cartilage and bone in vivo. This study was performed to investigate the effects of bone morphogenetic protein-2/4 on the characteristics of rat periodontal ligament cells(RPDL) and rat calvaria cells(RCV). In the control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 20% fetal bovine serum, $100{\mu}/ml$ penicillin, $100{\mu}/ml$ streptomycin. In the experimental groups, recombinant human bone morphogenetic protein-2/4 (25ng, 100ng, 250ng/ml) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 2, 3, 5, 7th day, the amount of total protein synthesis and alkaline phosphatase activity at 2, 5, 7th day. And also, the calcified nodule was examed. The results were as follows ; 1 . Both RCV and RPDL cells in both control and experimental groups proliferated during the entire experimental period, but there is no stastically significant difference according to the BMP-2/4 concentration. 2 . Amount of total protein synthesis of both cells in both groups was steadily increased until 5th day, but all experimental groups were significantly different from the control group at 7th day. 3. Alkaline phosphatase activity of both cells in both groups was increased during the entire experiment period. In RCV cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 7th day. In RPDL cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 5th day, and all experimental groups were significantly different from the control group at 7th day. 4. In the both of the cultured Rat Periodontal ligament and calvaria cell treated with BMP-2/4 to compared with control group, it revealed more rapid cell polarization, cell aggregation and hyperchromatic stained on HE agent, and even though only 1 day treated with BMP-2/4 both RPDL and RCV showed more rapid cell reaction than control group. More sensivitve cell reaction of RCV were observed than RPDL in this experiment. From the above results, we could conclude that BMP-2/4 influenced the induction, proliferation and differentiation of bone forming cells

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