• Journal of Periodontal and Implant Science
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• 제36권1호
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• pp.195-210
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• 2006

The aim of this study is to evaluate the effects of combination of rhBMP-4 and chitosan, which have osteoinductive capacity, on the regeneration of bone defects in dogs. Three beagle dogs aged over one and half years and weighed about 15Kg were used in this study. Three round defects were made by trephine bur in each side of mandibles. Each defect was filled with collagen, chitosan, rhBMP-4/chitosan. The dogs were sacrificed at 1, 3 or 8 weeks postsurgery and the results were evaluated histologically. The results of this study were as follows: 1. The sign of new bone formation, rearrangement of osteoblasts was revealed adjacent of preexisted bone or around graft materials such as chitosan. 2. The descending order of groups in bone regeneration speed was the rhBMP-4/chitosan group, chitosan group, collagen group. In the combination group, new bone was regenerated more and in better quality than others. 3. The regeneration of bone was observed in the rhBMP-4/chitosan group in 3 weeks after surgery. In 8 weeks after surgery, bone regeneration was observed in all three groups, and new bone at 8th week was denser in the chitosan and rhBMP-4/chitosan group than collagen group. In conclusion, rhBMP-4 and chitosan can be applied in the bone regeneration procedures usefully. It is considered that chitosan can be a splint for the maintenance of the defect form and produce much better effect when used with good grafting material or bone morphogenetic protein.

• 구강회복응용과학지
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• 제40권1호
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• pp.13-23
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• 2024

목적: 인간간엽줄기세포를 연골세포로 분화유도하는 성장인자 규명 및 주사침 크기 차이에 따른 세포증식 증진 비교이다. 연구 재료 및 방법: 인간간엽줄기세포를 연골세포유도배지에서 14, 21, 28일 배양하여 BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1. TGF-β2, TGF-β3를 투여한 군들과 대조군에서 SOX-5, SOX-6, SOX-9 및 FOXO1A의 발현량을 분석하였다. 18, 21, 26 게이지(G) 주사침을 사용하여 배양접시에 인간간엽줄기세포를 접종하고 24, 48, 72시간 후 각각 세포증식을 측정하였다. 결과: 기존에 알려진 FGF, IGF-1, TGFβ1와 함께, BMP-2, BMP-4, BMP-6, BMP-7 등 BMP family 성장인자들에 의하여 연골세포분화 전사인자 유전자들인 SOX-5, SOX-6, SOX-9, FOXO1A의 유전자발현이 증가하였다. 48시간에서는 가장 작은 주사침인 26G군이 대조군 및 18G군에 비하여 유의한 세포증식 증진을 보였으며, 72시간에서도 가장 작은 주사침인 26G군이 대조군에 비하여 유의한 세포증식 증진을 보였다. 결론: 인간간엽줄기세포의 연골세포분화 유도능을 가진 성장인자들을 고찰할 수 있었고 주사침 크기에 따른 세포증식 변화를 규명할 수 있었다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권4호
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• pp.316-322
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• 2004

A low molecular weight component named bone morphogenetic protein(BMP) chemically isolated from the organic matrix of bone, induce postfetal connective tissue cells surrounding small blood vessels to differentiate into cartilage and bone. The end product of BMP is a spherical ossicle of lamella bone filled with red bone marrow for the functional loading. This is a important point that the graft material is embedded the defect site during the implant surgery. Because present knowledge of the relationship between BMP and bone regeneration arises mainly from studies of induced bone formation in heterotopic sites, it would be helpful to determine whether BMP plays any part in the process of bone healing. The BMPs have been shown to play crucial roles in normal skeletal development as well as bone healing and are able to activate transcription of genes involved in cellular migration, proliferation, and differentiation. The delivery of BMP on matrices has been efficacious in the treatment of defect bone in implant surgery. The purpose of the histologic study was to evaluate the effect of DLB(demineralized lyophilized bone) coated with purified human BMP(hBMP-I) in immediate implant surgery with bony defect to obtain the functional structure of implant asap. The ability of a graft of hBMP-I to accelerate bony defect repair provides a rationale for its use in immediate implant surgery that have large bone defect in edentulous area.

• 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.

• BMB Reports
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• 제45권4호
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• pp.247-252
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• 2012

Although previous studies have demonstrated that BMP9 is highly capable of inducing osteogenic differentiation of mesenchymal stem cells, the molecular mechanism involved remains to be fully elucidated. In this study, we showed that BMP9 simultaneously promotes the activation of Smad1/5/8, p38 and ERK1/2 in C3H10T1/2 cells. Knockdown of Smad4 with RNA interference reduced nuclear translocation of Smad1/5/8, and disrupted BMP9-induced osteogenic differentiation. BMP9-induced osteogenic differentiation was blocked by p38 inhibitor SB203580, whereas enhanced by ERK1/2 inhibitor PD98059. SB203580 decreased BMP9-activated Smads singling, and yet PD98059 stimulated Smads singling in C3H10T1/2 cells. The effects of inhibitor were reproduced with adenovirus expressing siRNA targeted p38 and ERK1/2, respectively. Taken together, our findings revealed that Smads, p38 and ERK1/2 are involved in BMP9-induced osteogenic differentiation. Also, it is noteworthy that p38 and ERK1/2 may play opposing regulatory roles in mediating BMP9-induced osteogenic differentiation of C3H10T1/2 cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제47권6호
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• pp.454-464
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• 2021

Objectives: This study aimed to investigate the in vitro osteoinductivity of the combination of bone morphogenetic protein-2 (BMP-2) and nanohydroxyapatite (nHAp) and the in vivo effects of implants coated with nHAp/BMP-2. Materials and Methods: To evaluate the in vitro efficacy of nHAp/BMP-2 on bone formation, bone marrow-derived mesenchymal stem cells (BM-MSCs) were seeded onto titanium disks coated with collagen (Col), Col/nHAp, or Col/nHAp/BMP-2. Protein levels were determined by a biochemical assay and reverse transcriptase-polymerase chain reaction. Stem cell differentiation was analyzed by flow cytometry. For in vivo studies with mice, Col, Col/nHAp, and Col/nHAp/BMP-2 were injected in subcutaneous pockets. Titanium implants or implants coated with Col/nHAp/BMP-2 were placed bilaterally on rabbit tibias and evaluated for 4 weeks. Results: In the in vitro study, BM-MSCs on Col/nHAp/BMP-2 showed reduced levels of CD73, CD90, and CD105 and increased levels of glycosaminoglycan, osteopontin, and alkaline phosphatase activity. After 4 weeks, the Col/nHAp/BMP-2 implant showed greater bone formation than the control (P=0.07), while no differences were observed in bone implant contact and removal torque. Conclusion: These results suggest that a combination of BMP-2 and an nHAp carrier would activate osseointegration on dental implant surfaces.

• 한국콘텐츠학회논문지
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• 제10권3호
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• pp.196-203
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• 2010

본 연구는 BMP-4의 발현을 통해 골절 후 골유합에 대한 미세전류의 효과를 관찰하였다. 실험동물은 체중 2.5~3 kg내외의 6개월 령 뉴질랜드 웅성 토끼 24마리를 사용하였으며 경골 골절 후 미세전류를 적용한 실험군과 비적용군인 대조군으로 나누었고, 시간경과에 따른 변화를 관찰하기 위하여 3일, 7일, 14일 및 28일군으로 나누어 BMP-4에 대한 면역조직화학적 염색을 실행하여 다음과 같은 결론을 얻었다. BMP-4의 발현은 미세전류를 적용한 실험군과 자연치유군인 대조군 모두 시간이 경과함에 따라 증가하다가 감소되었다. 그러나 골절 7일 후 까지 동일 시점에서 실험군이 대조군에 비해 더욱 강한 면역양성 반응을 보였다. 특히 경골 골절 7일 후 대조군은 하버씨계의 동심원과 간질층판을 중심으로 중등도의 갈색의 면역양성반응을 보인데 반해 실험군의 경우 바깥층판을 포함하여 매우 강한 갈색의 면역양성반응을 보였다. 위의 결과로 보면 골절 후 미세전류를 적용할 때 치유과정 초기에 골형성단백질인 BMP-4의 발현을 증가시켜 골절 치유를 촉진시킴을 알 수 있었다.

• Journal of Periodontal and Implant Science
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• 제35권2호
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• pp.263-269
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• 2005

Bone morphogenetic protein(BMP) and platelet-derived growth factor(PDGF) have been demonstrated tostimulate bone formation when applied locally in vivo. To explore whether or not the combined use of BMP and PDGF could have promotive effect and synergic interaction on bone formation in vivo, bone marrow mesenchymal stem cells were treated with BMP-2, PDGF-BB, or BMP-2 plus PDGF-BB, and then these cells were injected into the subcutaneous space on the dorsum of nude mice. The bone formation was evaluated after 12 weeks. Histomorphometric analysis demonstrated that the subcutaneous nodules formed in nude mice contained 25.3% newly formed bone in the BMP-2 treated cells, 14.4% newly formed bone in the PDGF-BB treated cells, and 8.9% newly formed bone in the RMP-2 plus PDGF-BB treated cells. The results showed that the combination of BMP-2 and PDGF-BB had neither a promotive effect nor synergic interact on bone formation in vivo.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권4호
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• pp.412-418
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• 2008

Bone morphogenetic proteins (BMPs) in combination with stem cells gain more significance for their use in bone tissue engineering. The mesenchymal stem cell can be differentiated into osteoblast by the treatment of BMP. The aim of this study is to characterize the osteogenic differentiation process of adult stem cells derived from buccal fat pad according to BMP-2 within culture media and decide the appropriate concentration of BMP-2 to facilitate osteogenesis. The authors procured the stem cell from buccal fat pad and analyzed for presence of stem cell by flow cytomety against CD-34, CD-105 and STRO-1. The buccal fat derived stem cells (BFDC) were treated by application of the different concentration with BMP-2 of 0, 10, 50, 100 and 200ng/ml, respectively. And their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase(ALP) staining, Alizarin red staining and RT-PCR for osteocalcin(OC) gene expression at 7, 14 and 21day of culture. Flow cytometric analysis and biochemical assays demonstrated that BFDC might be a distinguished stem cells, and mineralization was accompanied in proportion to BMP-2 concentration. However, with 100ng/ml concentration of BMP-2, the BFDC demonstrated most efficient staining pattern of ALP and Alizarin red. The feasibility of the osteogenic differentiation in the group of both 50ng/ml and 200ng/ml of BMP-2 showed similar activity and relatively weaker than that of 100ng/ml. These results suggest that the BMP-2 stimulate osteogenesis by BFDC effectively and that bone induction might be controlled through negative regulatory feedback in higher concentration.

• Journal of Periodontal and Implant Science
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• 제35권2호
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• pp.345-357
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• 2005

Prostaglandin plays a significant role in the local control of bone metabolism associated with periodontal disease. ${\Delta}^{12}-PGJ_2$ is a natural $PGD_2$ metabolite that is formed in vivo in the presence of plasma. It is known for ${\Delta}^{12}-PGJ_2$ to stimulate calcification in osteoblastic cells. Bone morphogenetic protein(BMP) stimulated osteoblastic differentiation in various types of cells and greatly enhanced healing of bony defects. The purpose of this study was to evaluate the effect of rhEMP-2 on ${\Delta}^{12}-PGJ_2$ induced osteoblastic differentiation and mineralization in vitro. A human osteosarcoma cells line Saos-2 were cultured. In the test groups, 10-7M of ${\Delta}^{12}-PGJ_2$ or mixture of 10-8M of ${\Delta}^{12}-PGJ_2$ and 100ng/ml of rhBMP-2 or 100ng/ml of rhEMP-2 were added to culture media. After 1 day, 2 days and 4 days of culture period, the cell number was measured. Alkaline phosphatase activity was measure at 3 days. Reverse transcription polymerase chain reaction(RT-PCR) was performed to determine the expression of mRNA of bone matrix protein at 8 hours, 1 day and 7 days. The ability to produce mineralized nodules in rat osteoblasts(MC3T3-E1) was evaluated at 21 days. The results were as follows : 1. rhEMP-2 or mixture of rhBMP-2 and ${\Delta}^{12}-PGJ_2$ inhibited cell proliferation of human osteosarcoma cells. 2. rhEMP-2 or mixture of rhBMP-2 and ${\Delta}^{12}-PGJ_2$ stimulated alkaline phosphatase activity significantly higher than ${\Delta}^{12}-PGJ_2$ alone. 3. rhBMP-2 or mixture of rhEMP-2 and ${\Delta}^{12}-PGJ_2$ stimulated mineralization compared to ${\Delta}^{12}-PGJ_2$ alone. 4. mRNA of alkaline phosphatase, BMP-2, cbfa 1, Type I collagen were detected in the group treated with ${\Delta}^{12}-PGJ_2$/rhBMP-2, rhBMP-2 alone, ${\Delta}^{12}-PGJ_2$ alone. These results show that mixture of ${\Delta}^{12}-PGJ_2$ and rhBMP-2 causes more bone formation than ${\Delta}^{12}-PGJ_2$ alone while the bone formation effects of mixture of ${\Delta}^{12}-PGJ_2$ and rhBMP-2 are less than those of rhBMP-2 alone. Further researches would be necessary to clarify the interactions of these agents.