• Maxillofacial Plastic and Reconstructive Surgery
• /
• 제27권4호
• /
• pp.307-314
• /
• 2005

After dental implant are planted into their bony site among the various growth factors associated with bone formation. BMP is expressed in the bone surrounding the implant fixture. By taking a close look at BMP2, BMP4 which are growth factors that take put in bone formation, its histologic features and radiographic bone healing patterns we would like to examine the mechanism of osseointegration. We randomly used 8 male and female house rabbit amd used diameter 5 mm height spiral shaped implants(Ostem, Korea) for animal use handled as a resorbable blast machined(RBM) surface and machined surface. 2group were formed and each group had RBM surface and machined surface implant or a simple bone cavity. After 3, 7, 14 and 28 days post surgery 2 objects were sacrificed from each group and histologic specimens were acquired. RT-PCR analysis was conducted and after H&E staining the extent of osseointegration was measured applying a histologic feature and histomorphometric analysis program. Quanitity one -4.41(Bio-Rad, USA) was used after scanning the PCR product image of the growth factors manifested in each group. According to the histomorphometric features the RBM, Machined surface group showed increased contact between bone and implant surface at 3, 7, 14 and 28 days after surgery. The BMP2 level increased in both experiment groups but remained unchanged in the contrast group. BMP4 levels stayed steady after the early post implantation period for RBM but showed decreased in the machined surface group and contrast group. The amount of contact between bone and implant surface increased with the passage of time. BMP2, BMP4 were expressed in both experimental group and contrast group. These growth factors play a role in osseointegration of implant.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제39권3호
• /
• pp.112-119
• /
• 2013

Objectives: This study investigated the question of whether adenoviral magnetofection can be a suitable method for increasing the efficacy of gene delivery into bone marrow stromal cell (BMSC) and for generation of a high level of bone morphogenic protein (BMP) secretion at a minimized viral titer. Materials and Methods: Primary BMSCs were isolated from C57BL6 mice and transduced with adenoviral vectors encoding ${\beta}$ galactosidase or BMP2 and BMP7. The level of BMP secretion, activity of osteoblast differentiation, and cell viability of magnetofection were measured and compared with those of the control group. Results: The expression level of ${\beta}$ galactosidase showed that the cell transduction efficiency of AdLacZ increased according to the increased amount of magnetic nanoparticles. No change in cell viability was observed after magnetofection with 2 ${\mu}L$ of magnetic nanoparticle. Secretion of BMP2 or BMP7 was accelerated after transduction of AdBMP2 and 7 with magnetofection. AdBMP2 adenoviral magnetofection resulted in up to 7.2-fold higher secretion of BMP2, compared with conventional AdBMP2-transduced BMSCs. Magnetofection also induced a dramatic increase in secretion of BMP7 by up to 10-fold compared to the control. Use of only 1 multiplicity of infection (moi) of magnetofection with adenoviral transduction of AdBMP2 or AdBMP7 resulted in significantly higher transgene expression compared to 20 moi of conventional adenoviral transduction. Conclusion: Magnetic particle-mediated gene transudation is a highly efficient method of gene delivery to BMSCs. Magnetofection can lower the amount of viral particles while improving the efficacy of gene delivery.

• Journal of Periodontal and Implant Science
• /
• 제35권3호
• /
• pp.747-760
• /
• 2005

Periodontal therapy has dealt primarily with attempts at arresting progression of disease. however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. Recombinant human bone morphogenetic protein-7(rhBMP-7) can differentiate the osteoprogenitor cells and induce bone formation. The purpose of this study was to evaluate the effect of BMP-7 on rat periodontal ligament cells differentiation, in vitro. In the control group, cells was cultured with DMEM media. In the experimental groups, cells were cultured with rhBMP-7 in concentration of 10, 25, 50 and 100 ng/ml. Each group was characterized by examining alkaline phosphatase activity at 3 and 5 days of culture and the ability to produce mineralized nodules of rat calvarial cells at 14 days of culture. Synthesis of type I collagen(COL-I), osteocalcin(OCN), and bone sialoprotein(BSP) was evaluated by RT-PCR at 7 days of culture. Activation of Smad proteins and p38 MAP kinase was determined by western blot analysis of the cell lysates. Alkaline phosphatase activity was significantly increased in the concentration of BMP-7 50 ng/ml and 100 ng/ml compared to the control(p<0.05). The mineralized bone nodule formation was greater with addition of 50 ng/ml and 100 ng/ml BMP-7 than the control(p<0.01). In 7 days' culture, the expressions of COL-I, BSP, and OCN was increased by BMP-7 in concentration of 10 $ng/ml{\sim}100$ ng/ml. In western blot analysis, BMP-7 treated culture cells expressed Smad 1,5,8 in dose-dependent manner, whereas BMP-7 did not activate phosphorylated form of p38 MAP kinase. These result suggested that BMP-7 stimulate rat periodontal ligament cells to differentiate toward osteoblast phenotype and increase bone matrix production by activation of BMP-Smad pathway.

• Journal of Nutrition and Health
• /
• 제51권1호
• /
• pp.23-30
• /
• 2018

Purpose: Runx2 (runt-related transcription factor 2), a bone-specific transcription factor, is a key regulator of osteoblast differentiation and its expression is induced by the activation of BMP-2 signaling. This study examined whether zinc modulates BMP-2 signaling and therefore stimulates Runx2 and osteoblast differentiation gene expression. Methods: Two osteoblastic MC3T3-E1 cell lines (subclones 4 as a high osteoblast differentiation and subclone 24 as a low osteoblastic differentiation) were cultured in an osteogenic medium (OSM) as the normal control, Zn-($1{\mu}M$ Zn) or Zn+($15{\mu}M$ Zn) for 24 h. The genes and proteins for BMP-2 signaling (BMP-2, Smad-1/p-Smad-1), transcription factors (Runx2, osterix), and osteoblast differentiation marker proteins were assessed. Results: In both cell lines, BMP-2 mRAN and protein expression and extracellular BMP-2 secretion all decreased in Zn-. The expression of Smad-1 (downstream regulator of BMP-2 signaling) and p-Smad-1 (phosphorylated Smad-1) also downregulated in Zn-. Furthermore, the expression of the bone-specific transcription factors, Runx2 and osterix, decreased in Zn-, which might be due to the decreased BMP-2 expression and Smad-1 activation (p-Smad-1) by Zn-, because Runx2 and osterix both are downstream in BMP-2 signaling. Bone marker gene expression, such as alkaline phosphatase (ALP), collagen type I (COLI), osteocalcin, and osteopontin were also downregulated in Zn-. Conclusion: The results suggest that a zinc deficiency in osteoblasts suppresses the BMP-2 signaling pathway via the suppression of Smad-1 activation, and this suppressed BMP-2 signaling can cause poor osteoblast differentiation.

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

• 한국정보과학회:학술대회논문집
• /
• 한국정보과학회 2002년도 봄 학술발표논문집 Vol.29 No.1 (B)
• /
• pp.409-411
• /
• 2002

Enterprise JavaBeans(EJB)에서 영속적인 데이터들을 표현하고 관리하는 엔티티빈의 두 가지 구현 방법은 Bean Managed Persistence(BMP)와 Container Managed Persistence(CMP)기법이있다. 이렇게 엔티티빈이 데이터 베이스와 연결되어 사용되기 때문에 생기는 특성이 있는데, 이런 특성들이 BMP 와 CMP를 사용하여 구현. 운용. 유지보수 및 재사용을 알 때 많은 차이점을 만든다. EJB를 사용하는 소프트웨어 산업계에서, 이런 차이점에 대한 연구가 부분적으로는 이루어지고 있지만, 체계적인 비교 평가는 아직 미흡한 실정이다. 본 논문에서는 BMP와 CMP를 이용해서 빈을 구현할 때 나타나는 각각의 장점과 단점을 몇 가지 측면에서 살펴본다. 그리고 그 차이를 비교함으로써 BMP 와 CMP 기법을 평가한다

• 생명과학회지
• /
• 제22권4호
• /
• pp.456-465
• /
• 2012

유전자 재조합 이형이합체 인간 뼈 형성촉진인자(rhBMP)들은 뼈 재생을 위한 조직공학연구에 중요한 요소들이다. 그러나 실제 뼈 재생에 관한 연구를 수행함에 이들을 이용하는 것은 뼈 형성 촉진인자들이 짧은 반감기를 가지며 비교적 고농도의 단백질을 사용해야 하기 때문에 그만큼 많은 비용이 소요된다는 문제점을 가지고 있다. 이러한 한계를 뛰어넘기 위하여, 본 연구에서는 단백질 전달 서열(PTD)이 융합된 인간 뼈 형성촉진인자 2와 7이 이형이합체 유전자 재조합 단백질(rhBMP2/7-PTD)을 발현하는 세포주를 확립하였다. 이형이합체의 형성은 BMP 2와 BMP 7 단백질 및 PTD 영역의 사이에 각각 4개의 glycine 아미노산 염기서열이 첨가될 수 있도록 하여 각 단백질의 folding이 자유롭도록 디자인하였다. 이렇게 개발된 세포주는 고농도의 rhBMP2/7-PTD을 지속적으로 발현하여 배양액 내로 분비함으로 조직공학용 연구 및 개발에 효율적으로 이용 할 수 있다. 이상의 세포주에서 발현된 rhBMP2/7-PTD 단백질은 뼈 세포분화 유도를 확인 할 수 있는 ALP 활성을 나타냄으로써 뼈 성장촉진 단백질로서 생물학적인 활성을 가지고 있음을 보였다. 본 연구의 결과로 개발된 rhBMP2/7-PTD 형질전환 세포주는 향후 뼈 조직 재생과 같은 연구에 중요하고 효과적인 도구로 이용될 수 있을 것으로 사료된다.

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

• Journal of Microbiology and Biotechnology
• /
• 제27권7호
• /
• pp.1281-1287
• /
• 2017

Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of $EC_{50}$ of 2.93 ng/ml.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제45권3호
• /
• pp.123-128
• /
• 2019

Demineralized dentin matrix (DDM) has been used as a recombinant human bone morphogenetic protein-2 (rhBMP-2) carrier in many clinical trials. To optimize the clinical safety and efficacy of rhBMP-2 with DDM, efforts have been made to improve the delivery of rhBMP-2 by 1) lowering the administered dose, 2) localizing the protein, and 3) prolonging its retention time at the action site as well as the bone forming capacity of the carrier itself. The release profile of rhBMP-2 that is associated with endogenous BMP in dentin has been postulated according to the type of incorporation, which is attributed to the loosened interfibrillar space and nanoporous dentinal tubule pores. Physically adsorbed and modified, physically entrapped rhBMP-2 is sequentially released from the DDM surface during the early stage of implantation. As DDM degradation progresses, the loosened interfibrillar space and enlarged dentinal tubules release the entrapped rhBMP-2. Finally, the endogenous BMP in dentin is released with osteoclastic dentin resorption. According to the postulated release profile, DDM can therefore be used in a controlled manner as a sequential delivery scaffold for rhBMP-2, thus sustaining the rhBMP-2 concentration for a prolonged period due to localization. In addition, we attempted to determine how to lower the rhBMP-2 concentration to 0.2 mg/mL, which is lower than the approved 1.5 mg/mL.