• Title/Summary/Keyword: Bone morphogenetic protein 15

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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.

Association of a Single Codon Deletion in Bone Morphogenetic Protein 15 Gene with Prolificacy in Small Tail Han Sheep

  • Guo, W.;Chu, M.X.;Deng, X.M.;Feng, J.D.;Li, Ning;Wu, Changxin
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1491-1495
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    • 2004
  • Small Tail Han Sheep has significant characteristics of high prolificacy and non-seasonal ovulatory activity and is an excellent local sheep breed in P. R. China. Recently a novel member of the transforming growth factor $\beta$ (TGF$\beta$) superfamily termed bone morphogenetic protein 15 (BMP15) was shown to be specifically expressed in oocytes and to be essential for female fertility. Therefore, BMP15 is a candidate gene for reproductive performance of Small Tail Han Sheep. The whole genomic nucleotide sequence of BMP15 gene in Small Tail Han Sheep was searched for polymorphisms by PCR-SSCP and direct sequencing, and only one polymorphism was found. The polymorphism was a result of a 3 base pair deletion, which eliminated a single Leu codon (CTT). The allelic frequencies for A (without deletion) and B (with a codon deletion) are 0.73 and 0.27 respectively. The effects of BMP15 genotype on litter size were evaluated using the least squares model. This indicated that there was a significant association between litter size of Small Tail Han Sheep and a deletion in BMP15 gene (p=0.02<0.05). Small Tail Han Sheep ewes with AA and AB genotype produce on average 0.5 and 0.3 more lambs per litter than those ewes with BB genotype.

DEVELOPMENT OF BONE REGENERATING MATERIAL USING BONE MORPHOGENETIC PROTEIN(rhBMP-2) AND BIORESORBABLE POLYMER (유전자재조합 인간 골형성단백2 및 생흡수성고분자를 이용한 골형성유도체의 개발)

  • Lee, Jong-Ho;Kim, Jong-Won;Ahn, Kang-Min;Kim, Kack-Kyun;Lee, Zang-Hee
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.4
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    • pp.325-331
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    • 1999
  • We tested the bone regenerating capacity and histologic response of bioresorbable matrix-type implant, which was made with Poly(lactide-co-glycolide)(PLGA) and bone apatite for the carrier of bone morphogenetic protein(BMP). The critical size defect of 8mm in diameter was created at the calvaria of SD rats(n=18), and repaired with polymer implant with $15{\mu}g$ of rhBMP-2(n=9) or without it(n=9). At 2 weeks, 1 months after implantation, the animals were sacrificed(3 animals at every interval and group) and histologically evaluated. The calvarial defect which was repaired with polymer with BMP healed with newly formed bone about 70% of total defect. But that without BMP showed only 0 to under 30% bony healing. Inflammatory response was absent in both group through the experimental period, but there's marked foreign body giant response though it was a little less significant in polymer with BMP group. As the polymer was resorbed, the space was infiltrated and replaced by fibrovascular tissue, not by bone. In conclusion, our formulation of bioresorbable matrix implant loaded with bone morphogenetic protein works good as a bone regenerating material. However, it is mandatory to devise our system to have better osteoinductive and osteoconductive property, and less multinucleated giant cell response.

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Evaluation the Effectiveness of Fibrinogen to Overcome Bone Radiation Damage (방사성골괴사 극복을 위한 피브린지지체의 효용성 평가)

  • Jung, Hong-Moon
    • Journal of the Korean Society of Radiology
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    • v.15 no.4
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    • pp.539-545
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    • 2021
  • Radiation therapy is accompanied by adverse radiation effective. In particular, it is accompanied by disorders of the vascular system. Therefore, oxygen and nutrient deficiency occurs in the regeneration area. Eventually, osteoradionecrosis is formed in this cellular environment. According to a precedent study, bone morphogenetic protein-2 is used to overcome osteoradionecrosis. The purpose of this study was to investigate the regeneration ability of osteoradionecrosis by treating bone-forming protein-2 on a fibrinogen scaffold which is a biomaterial that is frequently used for bone regeneration after irradiation of the rat head. In addition, the purpose of this study was to verify the bone regeneration effect from the eight weeks. According to the experimental results, in the calvarial defected model of the irradiated mouse, making bone-formation was obtained after 8 weeks rather than bone-formation period in the early 4 weeks. moreover, it was found that the regenerated bone formation of the fibrinogen scaffold is formed from the inside of the bone of the defect area.

Alteration of TGFB1, GDF9, and BMPR2 gene expression in preantral follicles of an estradiol valerate-induced polycystic ovary mouse model can lead to anovulation, polycystic morphology, obesity, and absence of hyperandrogenism

  • Asghari, Reza;Shokri-Asl, Vahid;Rezaei, Hanieh;Tavallaie, Mahmood;Khafaei, Mostafa;Abdolmaleki, Amir;Seghinsara, Abbas Majdi
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.3
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    • pp.245-254
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    • 2021
  • Objective: In humans, polycystic ovary syndrome (PCOS) is an androgen-dependent ovarian disorder. Aberrant gene expression in folliculogenesis can arrest the transition of preantral to antral follicles, leading to PCOS. We explored the possible role of altered gene expression in preantral follicles of estradiol valerate (EV) induced polycystic ovaries (PCO) in a mouse model. Methods: Twenty female balb/c mice (8 weeks, 20.0±1.5 g) were grouped into control and PCO groups. PCO was induced by intramuscular EV injection. After 8 weeks, the animals were killed by cervical dislocation. Blood serum (for hormonal assessments using the enzyme-linked immunosorbent assay technique) was aspirated, and ovaries (the right ovary for histological examinations and the left for quantitative real-time polymerase) were dissected. Results: Compared to the control group, the PCO group showed significantly lower values for the mean body weight, number of preantral and antral follicles, serum levels of estradiol, luteinizing hormone, testosterone, and follicle-stimulating hormone, and gene expression of TGFB1, GDF9 and BMPR2 (p<0.05). Serum progesterone levels were significantly higher in the PCO animals than in the control group (p<0.05). No significant between-group differences (p>0.05) were found in BMP6 or BMP15 expression. Conclusion: In animals with EV-induced PCO, the preantral follicles did not develop into antral follicles. In this mouse model, the gene expression of TGFB1, GDF9, and BMPR2 was lower in preantral follicles, which is probably related to the pathologic conditions of PCO. Hypoandrogenism was also detected in this EV-induced murine PCO model.

Effects of Ulmus davidiana Planch(Ulmaceae) on mineralization, bone morphogenetic protein-2, alkaline phosphatase, type I collagen and collagennase-1 in bone cells (유근피가 골세포의 mineralization, bone morphogenetic protein-2, alkaline phosphatase, type I collagen 및 collagennase-1에 미치는 영향)

  • Byun, You-seok;Yoon, Jong-hwa;Hwang, Min-seob;Kim, Kap-sung;Jo, Hyun-seog
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.13-22
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    • 2005
  • By extracting the sample of Ulmus davidiana Planch(Ulmaceae), which was known to have the protection of damaged organ and the anti-inflammation action, it was experimented whether it is available for the application of treatment of osteoporosis. In the previous experiment, the extracts from Ulmus davidiana Planch(Ulmaceae) were confirmed to inhibit Cathepsin K through treating the cell of long bone, which contains osteoclast. Through this, it is suggested that Ulmus davidiana Planch(Ulmaceae) can play a role of prodrug as an inhibitor of absorbing bone ash in the treatment of osteoporosis. In the present experiment, a research in vitro Ulmus davidiana Planch(Ulmaceae) on the growth and sensibilization of osteoblast in a state that induced osteosis by using the cell tissue of MC3T3-El pre-osteoblastic was conducted. As a result, it could be confirmed that Ulmus davidiana Planch(Ulmaceae) has the strengthening function by enhancing the dosage and the activity of ALP depending on the time. The dosage was observed at the minimum of $50{\mu}g/m{\ell}$ and the maximum of $150{\mu}g/m{\ell}$. The enhancement in bone morphogenetic protein-2 at $100{\mu}g/m{\ell}$ UD could be observed, and it also increased the concentration of ALP mRNA within the cell of MC3T3-El. At $60{\mu}g/m{\ell}$ UD which indicated a little increase in Type I collagen mRNA for a long time of culture. However, it was shown to sharply inhibit the expression of gene in the culture between 15-20 days. These results suggest that Ulmus davidiana Planch(Ulmaceae) has an influence upon bone metabolism through thje sensibilization of osteoblast. Therefore, it could be known that utilized Ulmus davidiana Planch(Ulmaceae) can be positively applied for the general disease of bone metabolism through future studies.

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High-Level Expression of Recombinant Human Bone Morphogenetic Protein-4 in Chinese Hamster Ovary Cells

  • PARK JUNHO;YU SUNGRYUL;YOON JAESEUNG;BAEK KWANGHEE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1397-1401
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    • 2005
  • Bone morphogenetic protein-4 (BMP-4) is a signaling homodimeric molecule that acts as a morphogen to influence cell fate in a concentration-dependent manner. The limited supply of a pure preparation of BMP-4, due to very low level of their expression in vivo, makes it difficult not only to study the biological activities of BMPs, but also to use them as a clinical tool. For a large-scale production of BMP-4, human BMP-4 cDNA was expressed in Chinese hamster ovary (CHO) cells by a recently development vector system, which confers position-independent stable expression of the foreign genes. The CHO cell line expressing recombinant human BMP-4 (rhBMP-4) at the level of $7\;{\mu}g/ml$ could be obtained after stepwise selection with methotrexate. This level of expression is about 70 times higher than those previously reported. The partially processed form of BMP-4 as well as mature form could be detected, when the aliquots of culture media were analyzed by Western blot. The glycosylation pattern and biological activity of the rhBMP-4 were determined by glycosidase treatment and the induction rate of alkaline phosphatase in mouse osteoblastic cells.

Role of Growth Differentiation Factor 9 and Bone Morphogenetic Protein 15 in Ovarian Function and Their Importance in Mammalian Female Fertility - A Review

  • Castro, Fernanda Cavallari de;Cruz, Maria Helena Coelho;Leal, Claudia Lima Verde
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.8
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    • pp.1065-1074
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    • 2016
  • Growth factors play an important role during early ovarian development and folliculogenesis, since they regulate the migration of germ cells to the gonadal ridge. They also act on follicle recruitment, proliferation/atresia of granulosa cells and theca, steroidogenesis, oocyte maturation, ovulation and luteinization. Among the growth factors, the growth differentiation factor 9 (GDF9) and the bone morphogenetic protein 15 (BMP15), belong to the transforming growth factor beta (TGF-${\beta}$) superfamily, have been implicated as essential for follicular development. The GDF9 and BMP15 participate in the evolution of the primordial follicle to primary follicle and play an important role in the later stages of follicular development and maturation, increasing the steroidogenic acute regulatory protein expression, plasminogen activator and luteinizing hormone receptor (LHR). These factors are also involved in the interconnections between the oocyte and surrounding cumulus cells, where they regulate absorption of amino acids, glycolysis and biosynthesis of cholesterol cumulus cells. Even though the mode of action has not been fully established, in vitro observations indicate that the factors GDF9 and BMP15 stimulate the growth of ovarian follicles and proliferation of cumulus cells through the induction of mitosis in cells and granulosa and theca expression of genes linked to follicular maturation. Thus, seeking greater understanding of the action of these growth factors on the development of oocytes, the role of GDF9 and BMP15 in ovarian function is summarized in this brief review.

Novel analysis model for implant osseointegration using ectopic bone formation via the recombinant human bone morphogenetic protein-2/macroporous biphasic calcium phosphate block system in rats: a proof-of-concept study

  • Park, Jung-Chul;Lee, Jong-Bin;Daculsi, Guy;Oh, Sang-Yeop;Cho, Kyoo-Sung;Im, Gun-Il;Kim, Byung-Soo;Kim, Chang-Sung
    • Journal of Periodontal and Implant Science
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    • v.42 no.4
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    • pp.136-143
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    • 2012
  • Purpose: The osseointegration around titanium mini-implants installed in macroporous biphasic calcium phosphate (MBCP) blocks was evaluated after incubation with recombinant human bone morphogenetic protein-2 (rhBMP-2) in an ectopic subcutaneous rat model. Methods: Mini-implants (${\varphi}1.8{\times}12$ mm) were installed in MBCP blocks (bMBCPs, $4{\times}5{\times}15$ mm) loaded with rhBMP-2 at 0.1 mg/mL, and then implanted for 8 weeks into subcutaneous pockets of male Sprague-Dawley rats (n=10). A histomorphometric analysis was performed, and the bone-to-implant contact (BIC) and bone density were evaluated. Results: Significant osteoinductive activity was induced in the rhBMP-2/bMBCP group. The percentage of BIC was $41.23{\pm}4.13%$ (mean${\pm}$standard deviation), while bone density was $33.47{\pm}5.73%$. In contrast, no bone formation was observed in the bMBCP only group. Conclusions: This model represents a more standardized tool for analyzing osseointegration and bone healing along the implant surface and in bMBCPs that excludes various healing factors derived from selected animals and defect models.

Effects of Bojungikgitang-gagambang on Longitudinal Bone Growth in Adolescent Rats

  • Lee, Min-Su;Kim, Ji-Young;Kim, Mi-Yeon;Chang, Gyu-Tae;Kim, Ho-Cheol
    • The Journal of Korean Medicine
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    • v.33 no.2
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    • pp.1-10
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    • 2012
  • Objectives: This study was aimed to investigate the effect of Bojungikgitang-gagambang (BJIG) on longitudinal bone growth in rats. Methods: The BJIG treated group (300 mg/kg) and the control group (vehicle) were administered orally twice daily for 4 days. To investigate the effects of BJIG we measured body weight gain. The bone growth effect was analyzed by measuring between fluorescent lines marked with tetracycline, which plays the role of fluorescent dye on the surface of the tibia. Tetracycline was intraperitoneally injected. The height of growth plates in the epiphyseal plate was measured. The expression of bone morphogenetic protein-2 (BMP-2) and insuline-like growth factor-1 (IGF-1) was investigated by immunohistochemistry. Results: BJIG caused a significant acceleration of longitudinal bone growth of $349.7{\pm}15.9{\mu}m/day$ compared to control ($319.8{\pm}21.4{\mu}m/day$). The height of overall growth plate was not significantly more compared to the control, but the size of cells in the proliferative zone and hypertrophic zone were. In the immunohistochemistry, BMP-2 and IGF-1 were expressed markedly in the proliferative or hypertrophic zone, respectively. Conclusions: BJIG stimulated the chondrocyte hypertrophy and chondrogenesis in the growth plate and directly increased the longitudinal tibia length of rats.