• Journal of Periodontal and Implant Science
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• 제37권4호
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• pp.859-869
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• 2007

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

• 유기물자원화
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• 제23권4호
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• pp.11-20
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• 2015

원지반에서의 저품위 석탄층의 생물학적 메탄 생산을 위한 기초연구를 수행하였다. 인도네시아산 갈탄을 시료로 이용하였으며, 조건을 달리하여 BMP(Biochemical Methane Potential) 실험을 수행하였다. 갈탄에 영양물질과 혐기성 슬러지만 제공한 경우, 온도(23, $30^{\circ}C$)나 석탄의 입자크기는 메탄가스 생산에 영향을 주지 않았다. 이는 가용한 용해성 유기물질이 낮기 때문이다. 외부탄소원으로 볏짚을 갈탄에 첨가한 후 BMP 실험을 수행하였으며, 60일간의 BMP 실험 후 볏짚 첨가에 의한 메탄가스 발생량은 94.4~110.4 mL/g VS이었다. BMP 실험 후 갈탄의 발열량은 볏짚 첨가량이 증가할수록 감소하는 경향을 보였다. 이는 볏짚의 혐기성 분해와 함께 갈탄의 분해가 이루어졌음을 의미한다. 따라서 원지반에서의 저품위 석탄층의 생물학적 메탄 생산의 초기 운전 시에 볏짚을 탄소원으로 사용할 수 있다.

• Clinical and Experimental Reproductive Medicine
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• 제34권4호
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• pp.239-252
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• 2007

목 적: 사람의 HAD와 HUC를 심근세포로 분화 유도하고자 하였다. 연구방법: 사람의 HAD와 HUC를 분리하여 5-azacytidine을 24시간 처리하고 여러 가지 BMP와 FGF을 첨가하여 배양하였다. 또한 HUC은 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1 또는 Wnt inhibitor를 첨가하여 배양한 후 심근세포 특이 유전자의 발현을 조사하였다. 결 과: HAD를 5-azacytidine 처리하고 기본배양액에서 4주 동안 배양하였을 때 TnT 유전자가 새로이 발현하였으며 Cmlc1과 kv4.3의 발현 양이 증가하였다. 5-azacytidine 처리 후에 BMP-4와 함께 FGF-4 (B4/F4) 또는 FGF-8 (B4/F8)을 첨가하여 배양하였을 때는 $\beta$-MHC 유전자 발현이 새로이 유도되었으며, Cmlc1, TnT, TnI 그리고 Kv4.3 유전자 발현 양이 더 많이 증가하였다. HUC은 5-azacytidine 및 BMP와 FGF 처리에 의해 유전자 발현 변화가 없었다. 그러나 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1을 첨가하여 배양하였을 때, BMP-2와 FGF-8 (B2/F8)을 첨가하여 배양한 세포에서 $\beta$-MHC 발현이 새로이 유도되었으며 $\alpha$-CA, TnT 그리고 Kv4.3 유전자의 발현이 증가하였다. 또한 BMP와 FGF와 함께 Wnt inhibitor를 처리하여 1주 동안 배양하였을 때 Cinlc1 유전자 발현이 새로이 유도되었으며 $\alpah$-CA, TnT, TnI 그리고 Kv4.3의 발현이 증가되었다. 결 론: HAD는 BMP와 FGF 처리에 의해 심근세포 특이 유전자의 발현증가를 유도할 수 있었으며 HUC는 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1 또는 Wnt inhibitor를 처리함으로써 심근세포 특이 유전자의 발현증가를 유도할 수 있었다. 따라서 HAD와 HUC는 심장질환 치료를 목적으로 하는 세포 치료에 이용될 수 있을 것으로 사료된다.

• International Journal of Stem Cells
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• 제16권4호
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• pp.406-414
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• 2023

Dedifferentiated fat cells (DFATs) isolated from mature adipocytes have a multilineage differentiation capacity similar to mesenchymal stem cells and are considered as promising source of cells for tissue engineering. Bone morphogenetic protein 9 (BMP9) and low-intensity pulsed ultrasound (LIPUS) have been reported to stimulate bone formation both in vitro and in vivo. However, the combined effect of BMP9 and LIPUS on osteoblastic differentiation of DFATs has not been studied. After preparing DFATs from mature adipose tissue from rats, DFATs were treated with different doses of BMP9 and/or LIPUS. The effects on osteoblastic differentiation were assessed by changes in alkaline phosphatase (ALP) activity, mineralization/calcium deposition, and expression of bone related genes; Runx2, osterix, osteopontin. No significant differences for ALP activity, mineralization deposition, as well as expression for bone related genes were observed by LIPUS treatment alone while treatment with BMP9 induced osteoblastic differentiation of DFATs in a dose dependent manner. Further, co-treatment with BMP9 and LIPUS significantly increased osteoblastic differentiation of DFATs compared to those treated with BMP9 alone. In addition, upregulation for BMP9-receptor genes was observed by LIPUS treatment. Indomethacin, an inhibitor of prostaglandin synthesis, significantly inhibited the synergistic effect of BMP9 and LIPUS co-stimulation on osteoblastic differentiation of DFATs. LIPUS promotes BMP9 induced osteoblastic differentiation of DFATs in vitro and prostaglandins may be involved in this mechanism.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권4호
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• pp.282-291
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• 2004

Pure-phase beta-tricalcium phosphate(${\beta}$-TCP) proved to be a bone regeneration material, providing the patient with vital bone at the defect site in a reasonable time, making a second surgical procedure for bone harvesting unnecessary. This study compares bone healing and BMP 2/4 expression in cranial defects in rabbits grafted with autogenous bone and ${\beta}$-TCP. Thirty New Zealand White rabbits was divided into 3 group of 10 animals each. Bilateral calvarial defects were made in the parietal bones of each animal. ${\beta}$-TCP placed in one defect and the other defects was filled with autogenous bone. The animal were sacrificed at 4, 8 and 12 weeks. Immunohistochemical analysis was used to investigate the expression of BMP 2/4. 1. The new bone formation around autogenous bone from 4 weeks and ${\beta}$-TCP from 8 weeks. 2. In autogenous bone graft, BMP 2/4 expression was decreased from 4 to 12 weeks. 3. In ${\beta}$-TCP graft, BMP 4 expression was increased from 8 to 12 weeks. But, BMP 2 was observed from 12 weeks. This study showed that bone healing, regeneration and, BMP 2/4 expression are delayed in grafted ${\beta}$-TCP than autogenous bone.

• International Journal of Oral Biology
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• 제32권3호
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• pp.103-107
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• 2007

We hypothesized that plaque-associated bacteria may have a role in maintenance of alveolar bone. To test it, immortalized gingival epithelial HOK-16B cells were co-cultured with live or lysed eight plaque bacterial species and the expression levels of bone morphogenetic protein (BMP)-2 and -4 were examined by real time reverse transcription-polymerase chain reaction. Un-stimulated HOK-16B cells expressed both BMP-2 and -4. Co-culture with plaque bacterial lysates had significant effects on the level of BMP-2 but not on that of BMP-4. Five species including Streptococcus sanguinis, S. gordonii, Veillonella atypica, Porphyromonas gingivalis, and Treponema denticola substantially up-regulated the level of BMP-2. In contrary to the upregulatory effect of lysate, live T. denticola suppressed the expression of BMP-2. In addition, in vitro osteoblastic differentiation assay using C2C12 cells and the conditioned medium of HOK-16B cells confirmed the production of BMPs by gingival epithelial cells and the modulation of BMP expression by the lysates of S. sanguinis and T. denticola. In conclusion, we have shown that plaque bacteria can regulate the expression of BMP-2 by gingival epithelial cells, the physiologic meaning of which needs further investigation.

• 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.

• 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.

• 한국수자원학회:학술대회논문집
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• 한국수자원학회 2016년도 학술발표회
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• pp.565-565
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• 2016

농림부는 농업지역에서 발생하는 비점오염원을 관리하기 위해 다양한 최적관리기법(BMP)에 대해 연구를 수행하여 효과를 검증하였으나 이를 광역 단위로 보급하여 실제 영농지역에서 BMP의 효과를 측정한 연구는 부족한 실정이다. 따라서 본 연구에서는 논 지역을 대상으로 물관리기술(물꼬)과 시비관리기술(완효성 비료) BMP를 보급하고 광역 단위로 BMP의 효과를 측정하고자 하였다. 연구대상지구는 새만금 지역에 위치한 전라북도 부안군 용계리 일대에 위치하고 있으며, 벼농사가 활발하게 이루어지고 있는 지역을 선정하였다. 논에 보급된 BMP의 효과를 측정하기 위해 BMP 참여 농가를 선정하여 물꼬와 완효성 비료를 보급하였으며, 용수로와 배출구 3지점을 선정하여 관개량을 측정하고 유출량을 측정하였다. 또한 관개수와 대조구, 처리구별(대조구, 물꼬, 완효성, 물꼬+완효성) 수질을 분석하여 배출구 별로 단위면적당 오염부하량을 산정하고 이를 통해 대조구 대비 처리구의 저감율을 분석하였다. 분석 결과, 대조구 대비 처리구의 단위면적당 오염부하량의 저감율은 SS 63.6%, BOD 45.5%, $COD_{Mn}$ 26.5%, $COD_{Cr}$ 40.1%, T-N 16.0%, T-P 12.9%, TOC 11.4%로 나타났다. 또한 연구대상지구를 SO#1(완효성 비료 95%, 비참여 농가 5%), SO#2(완효성 비료 2%, 물꼬 8%, 완효성 비료+물꼬 40%, 비참여 농가 40%), SO#M(완효성 비료 96%, 비참여 농가 4%) 3개의 배출구로 구분하여 BMP 보급 효과를 측정한 결과, 대조구 대비 SO#1에서는 수질항목별로 10.6 ~ 85.5%, SO#2는 8.1 ~ 45.9%, SO#M은 10.7 ~ 86.2%의 범위로 저감된 것으로 분석되었으며, 특히 SS의 단위면적당 오염부하 저감율이 가장 큰 것으로 나타났다. 하지만 본 연구는 7월부터 10월까지 측정된 데이터를 활용하여 산정하였기 때문에 써레질, 이앙기간에 발생한 단위면적당 오염부하량은 포함되지 않은 결과이며, 각 배수구역별 유입량의 경우 정확한 측정에 어려움이 있어 전체 관개량 대비 관개 면적을 이용하여 단위면적당 유입량을 산정하였기 때문에 추가적인 연구를 통해 효과를 검증할 필요가 있을 것으로 판단된다.

• 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.