• 생명과학회지
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• 제27권5호
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• pp.501-508
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• 2017

Bone morphogenetic proteins (BMPs)는 다양한 세포기능을 조절하는 중요한 사이토카인 중 하나이다. 최근 BMP와 일주기 유전자들이 연관되어 있다는 연구결과들이 보고되고 있지만 조골세포에서 일주기 유전자인 Per1의 역할은 아직 명확하지 않다. 본 연구에서는 조골세포 분화에서 Per1의 역할을 조사하였다. MC3T3-E1 세포에서 BMP2 처리에 의해 Per1 mRNA 발현과 luciferase 활성이 증가하는 것을 확인하였다. 또한 Per1 과발현 실험을 통해서 Per1 유전자가 Runx2, ALP, OC의 발현을 증가시켰으며 ascorbic acid와 ${\beta}$-glycerophosphate에 의한 ALP 염색과 석회화가 Per1 과발현에 의해 더욱 증가하는 것을 확인하였다. 이상의 결과는 일주기 리듬을 조절하는 Per1 유전자가 조골세포의 분화를 촉진하는 인자로 작용함을 시사한다.

• 생약학회지
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• 제45권4호
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• pp.326-331
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• 2014

In this study, we investigated pharmacologic activity of rice bran ethyl acetate fraction (RBE), based on their osteoblast enhancing effects. It has been found that REB have a stimulatory effect on the commitment of bi-potential mesenchymal precursor C2C12 cells into osteoblasts in the presence of BMP-2. Furthermore, RBE enhanced the BMP-2-stimulated induction of ALP, an early phase biomarker of osteoblast differentiation. In addition, Western blot analysis showed RBE enhanced the BMP-2-stimulated phosphorylation of p38, but not those of ERK or JNK. These findings show RBE has the potential to enhance the BMP-2-mediated commitment of C2C12 cells into osteoblasts and their differentiation through p38 activation.

• Journal of Periodontal and Implant Science
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• 제49권2호
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• pp.114-126
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• 2019

Purpose: The aim of this study was to evaluate the enhancement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with Escherichia coli-derived recombinant human bone morphogenetic protein-2 (ErhBMP-2) and epigallocatechin-3-gallate (EGCG). Methods: The cell viability, differentiation, and mineralization of osteoblasts was tested with ErhBMP-2-/EGCG solution. Coated BCP surfaces were also investigated. Standardized, 6-mm diameter defects were created bilaterally on the maxillary sinus of 10 male New Zealand white rabbits. After removal of the bony windows and elevation of sinus membranes, ErhBMP-2-/EGCG-coated BCP was applied on one defect in the test group. BCP was applied on the other defect to form the control group. The animals were sacrificed at 4 or 8 weeks after surgery. Histologic and histometric analyses of the augmented graft and surrounding tissue were performed. Results: The 4-week and 8-week test groups showed more new bone (%) than the corresponding control groups (P<0.05). The 8-week test group showed more new bone (%) than the 4-week test group (P<0.05). Conclusions: ErhBMP-2-/EGCG-coated BCP was effective as a bone graft material, showing enhanced osteogenic potential and minimal side effects in a rabbit sinus augmentation model.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7091-7095
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• 2014

Background: The study aimed to evaluate the incidence of CpG island promoter methylation of BMP6, a member of the transforming growth factor beta family, in tissue samples from colorectal cancers (CRC) and look for its association with BMP6 expression and clinicopathological correlation. Materials and Methods: Methylation specific PCR for the BMP6 promoter region was performed with 85 frozen tissue samples of CRC and 45 of normal colon. Methylation status of MLH1 was also determined by the same method. Expression of BMP6 was evaluated by immunohistochemistry (IHC), using Allred's scoring system. The methylation status was analyzed against clinical and pathological parameters in CRC. Results: The study revealed BMP6 hypermethylation in 34 of 85 tumor specimens (40%), and 15 out of 45 normal tissue samples from CRC (33%). The incidence of hypermethylation was inversely correlated with IHC score. Allred's scores of 7 or more were correlated with lower frequency of BMP6 hypermethylation (29% compared to 50% in the remaining, p-value 0.049). However, there was no association between hypermethylation status and any clinicopathological parameters. The methylation status of BMP6 was not correlated with that of MLH1, a key methylation determinant in CRC. On survival analysis, there was no significant difference in progress-free survival (PFS) between the cases with and without hypermethylation (2-year PFS 74% and 76%, respectively). Conclusions: CpG island methylation of BMP6 is found in high frequency in CRC and this epigenetic event is associated with suppressed protein expression in the tumor tissue. However, the marker is not associated with tumor progression of the disease.

• 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.

• 대한환경공학회지
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• 제36권8호
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• pp.521-527
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• 2014

본 연구의 목적은 필지 단위 최적관리기법 적용 효과를 분석할 수 있는 HRSM4BMP 모형을 유역단위로 모의할 수 있도록 모형을 개선하는 것이며 이를 실제 유역에 적용하여 평가하는 것이다. 따라서 HRSM4BMP 모형을 유역단위로 모의할 수 있도록 하기 위해 전처리 프로세서와 후처리 프로세서 모듈을 개발하였고 유역단위 유출량 및 유사량 모의에 많이 사용되고 있는 SWAT모형의 하천 하도추적과정과 HRSM4BMP 모형을 연계하였다. 이렇게 개선된 HRSM4BMP 모형을 강원도 양구군 해안면 유역 내 14번 소유역에 2 m의 초생대 최적관리기법 적용에 따른 유역단위 저감효과를 모의하였다. 그 결과, 14번 소유역에서는 약 280 ton (5%)의 유사가 저감되었으며 유역 말단인 43번 소유역에서 초생대 적용 후 약 223 ton (4%)의 저감 효과가 있는 것으로 분석되었다. 본 연구의 결과는 향후 우리나라 유역단위 BMPs 설치 장기 계획에 있어 매우 유용하게 사용될 수 있을 것이라 판단된다.

• 한국임상수의학회지
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• 제22권4호
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• pp.377-381
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• 2005

We tried to extract bone morphogenetic protein (BMP) from the freeze-dried bovine cortical bone (FBCB) for bone graft, which were defatted with chloroform-methanol for 20 days, freeze-dried at $-80^{\circ}C$ for 7 days and sterilized by ethylene oxide gas. Two kg of FBCB were pulverized in a wheel mill to $0.5-2.0mm^3$ cubic in size. The bone particles were demineralized in 0.6N HCI for 10 days at chloroform-methanol$4^{\circ}C$ and defatted with chloroform-methanol for 6 hours at room temperature, which was going to be defatting and demineralized cortical bone (DDM). For extracting BMP, DDM was agitated continuously through 72 hours with magnetic stirrer at $4^{\circ}C$ into 12 times of volume of 6 M guanidine hydrochloride (Gdn-HCl) solution containing proteinase inhibitors to protect BMP such as 2mM N-ethylaleimide, 1mM iodoacetic acid, 1mM phenylmethylsulfonyl fluoride and a sterilizer, 1mM sodium azide. The extraction procedure was repeated for three times. All extracted solution was centrifuged at 10,000 rpm for 30 min and then, the supernatant was dialyzed with 12 times of volume of deionized water at $4^{\circ}C$ for 24-72 hours, which cut off below 6,000-8,000 molecular weight. The dialyzed specimen contained crude-BMP was centrifuged, freeze-dried, and weighted. Through these processing, we could obtained $84.9\%$ as FBCB, $17.8\%$ as DDM and $0.71\%$ as crude-BMP from the wet cortical bone without cancellous bone, marrow and muscles. The crude-BMP were obtained $68.3\%$ from the first extraction, $29.6\%$ from secondary and $2.1\%$ from tertiary, respectively. It was suggested that high yield of crude-BMP migth be explained by three-time repetition of the extraction processing for crude-BMP with Gdn-Hcl sol.

• Journal of Periodontal and Implant Science
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• 제26권1호
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• pp.68-79
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• 1996

To investigate the efficiency of a fibrous collagen membrane(FCM) composed of bovine skin type I atelocollagen as a carrier for BMP, partially purified bovine BMP/FCM($0.3mg/10{\times}5{\times}1mm$) composites were implanted into the dorsal subcutaneous tissue of rats. FCM alone was also implanted as a control. The implants were harvested at 1, 2, 3, and 10 weeks after implantation, then prepared for routine light microscopic observation. The FCM alone did not induce osteogenesis and revealed no specific foreign body reaction nor was there any definite resorptive evidence for 10 weeks after implantation, while the BMP/FCM composites induced favorable bone formation in a process that resembled an endochondral and direct ossification mode. At 10 weeks, the well formed bone confined to embedded collagen fibers revealed hematopoietic marrow between bony trabeculae without evidence of resorptive or degenerative changes . These findings support the suggestion that BMP may induce undifferentiated mesenchymal cells into either chondroblasts or osteoblasts or both independantly according to the chemico- physical characteristics of the carrier, which develops the endochondral and/or direct bone formation process, and suggest that the FCM may be a favorable carrier for BMP.

• Journal of Periodontal and Implant Science
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• 제36권3호
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• pp.627-637
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• 2006

이 연구는 rh BMP-7-immobilized substrates에 대한 백서 태자 두개관 세포의 반응을 석회화 결절 측정, 알카리 효소 분석, 역전사 중합반응 및 단백질 분석등으로 평가하여 다음과 같은 결과를 얻었다. 1. 배양 14일 째, 석회화 결절 형성율을 측정한 결과, rh BMP-7-immobilized substrates에서 대조군과 비교하여 더 많은 석회화 결절을 형성하였다. 2. 배양 7일에 염기성 인산 분해효소 활성도는 rh BMP-7-immobilized substrates에서 대조군에 비해 효소 활성도가 유의하게 높았다. 3. 역전사 중합반응의 결과에서 BSP 와 OCN 유전자 발현은 대조군보다 더 현저하였다. 4. 단백질 분석에서 rh BMP-7-immobilized substrates와 대조군 모두 Smad 1,5,8 단백질의 인산화를 활성화시키지 못했다. 이상의 결과 rh BMP-7-immobilized substrates는 백서 태자 두개관세포가 조골세포로의 분화와 석회화를 유도하며 따라서 rh BMP-7-immobilized substrates는 임프란트 주변의 골 형성에 유용하리라 사료된다.

• BMB Reports
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• 제49권3호
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• pp.179-184
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• 2016

Bone morphogenetic protein 9 (BMP9) is a potent inducer of osteogenic differentiation of mesenchymal stem cells. The Wnt antagonist Dickkopf-1 (Dkk1) is involved in skeletal development and bone remodeling. Here, we investigated the role of Dkk1 in BMP9-induced osteogenic differentiation of MSCs. We found that overexpression of BMP9 induced Dkk1 expression in a dose-dependent manner, which was reduced by the P38 inhibitor SB203580 but not the ERK inhibitor PD98059. Moreover, Dkk1 dramatically decreased not only BMP9-induced alkaline phosphatase (ALP) activity but also the expression of osteocalcin (OCN) and osteopontin (OPN) and matrix mineralization of C3H10T1/2 cells. Furthermore, exogenous Dkk1 expression inhibited Wnt/β-catenin signaling induced by BMP9. Our findings indicate that Dkk1 negatively regulates BMP9-induced osteogenic differentiation through inhibition of the Wnt/β-catenin pathway and it could be used to optimize the therapeutic use of BMP9 and for bone tissue engineering.