• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.33 no.4
• /
• pp.288-296
• /
• 2007

Distraction osteogenesis(DO) is a technique of lengthning bone including soft tissue by gradual separation of surgically divided bone surfaces. Distraction osteogenesis combination with a compression stimulation(DO-CO) was a new technique by authors to enhance new bone quality and to shorten the consolidation period. The purpose of this study was to compare DO with DO combined with compression force in efficiency by evaluating the expression of TGF-${\beta}1$, osteonectin and BMP-4 on bone regenerate in rabbit mandible. Fourty two rabbits were used for this experiment. On the control group, the distraction was carried out at the rate of 1 mm per day to obtain the amount of 8 mm distraction for 8 days. On the experimental group, the distraction was carried out at the rate of 1 mm per day for 10 days, 3 days-latency period, and then the compression was carried out as counter direction 1 mm per day for 2 days. After 0 day, 5 days, 13 days, 20 days, 27 days, 34 days and 41 days, three rabbits on each group were sacrificed and the distracted portion of mandible were cut and treated for RT-PCR observation. The level of expression of TGF-${\beta}1$ and osteonectin were shown more and longer expression in the experimental group than in the control group. The expression of BMP-4 was maintained with high level during the entire experimental period in both groups. These findings suggested that DO with compression stimulation could be a favorable technique for obtaining a good new bone quality.

• Proceedings of the Korea Contents Association Conference
• /
• 2009.05a
• /
• pp.1124-1129
• /
• 2009

This study aims to examine the effects of non-invasive constant microcurrent stimulation on expression of Bone Morphogenetic Protein(BMP) 4 after tibia fracture in rabbits. Twenty four rabbits with tibia fracture were randomly divided into control and experimental groups. Each group was divided into four subgroups, based on the duration of the experiment (3, 7, 14, 28 days). The experimental groups received a constant microcurrent stimulation of $20{\sim}25{\mu}A$ intensity with surface Ag-AgCl electrode (diameter 1cm, Biopac, U.S.A.) for 24 hours a day. Cathode of the microcurrent stimulator located on the tibia directly, anode of it did on the gastrocnemius muscle. Rabbits were sacrificed on each of the postoperative days 3, 7, 14, 28. To investigate how non- invasive constant microcurrent stimulation affects bone healing, immunohistochemical analysis of BMP-4 was performed at each point. After evaluation, the test results are as follows: Comparisons of immunohistochemical observation of BMP-4 in 7 days after tibial fracture show that there was shown to be a moderate positive reaction (++) on concentric circles of Harversian system andt he interstitial lamella in the control group, while there was a very strong positive reaction (++++) on concentric circles of Harversian system and interstitial lamellain the experimental group. These results suggest that applying non-invasive constant microcurrent stimulation on fractured bone is helpful to bone healing.

• Molecules and Cells
• /
• v.39 no.5
• /
• pp.389-394
• /
• 2016

Dovitinib (TKI258) is a small molecule multi-kinase inhibitor currently in clinical phase I/II/III development for the treatment of various types of cancers. This drug has a safe and effective pharmacokinetic/pharmacodynamic profile. Although dovitinib can bind several kinases at nanomolar concentrations, there are no reports relating to osteoporosis or osteoblast differentiation. Herein, we investigated the effect of dovitinib on human recombinant bone morphogenetic protein (BMP)-2-induced osteoblast differentiation in a cell culture model. Dovitinib enhanced the BMP-2-induced alkaline phosphatase (ALP) induction, which is a representative marker of osteoblast differentiation. Dovitinib also stimulated the translocation of phosphorylated Smad1/5/8 into the nucleus and phosphorylation of mitogen-activated protein kinases, including ERK1/2 and p38. In addition, the mRNA expression of BMP-4, BMP-7, ALP, and OCN increased with dovitinib treatment. Our results suggest that dovitinib has a potent stimulating effect on BMP-2-induced osteoblast differentiation and this existing drug has potential for repositioning in the treatment of bone-related disorders.

• Journal of Periodontal and Implant Science
• /
• v.36 no.3
• /
• pp.627-637
• /
• 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.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.40 no.1
• /
• pp.13-23
• /
• 2024

Purpose: This aim of this study was to demonstrate growth factors that differentiate human mesenchymal stem cells into chondrocytes and to evaluate cell proliferation enhancement by needle size differences. Materials and Methods: Human mesenchymal stem cells were cultured in chondrogenic medium supplemented with BMP-2, BMP-4, BMP-6, BMP-7, BMP-13, FGF-2, FGF-18, IGF-1, TGF-β1, TGF-β2, TGF-β3 and without growth factors for 14, 21, and 28 days. Then, the expression levels of SOX-5, SOX-6, SOX-9 and FOXO1A were comparatively analyzed. Human mesenchymal stem cells were inoculated into culture dishes using 18, 21, and 26 gauge (G) needles, and cell proliferation was measured after 24, 48, and 72 hours, respectively. Results: In addition to the previously known FGF, IGF-1, and TGFβ1,the BMP family growth factors such as BMP-2, BMP-4, BMP-6, and BMP-7 increased the expression of chondrocyte differentiation genes SOX-5, SOX-6, SOX-9, and FOXO1A. At 48 hours, the 26G group, the smallest needle, showed significant cell proliferation improvement compared to the control group and the 18G group. At 72 hours, the 26G group, the smallest needle, showed significant increase in cell proliferation compared to the control group. Conclusion: Through this study, growth factors with the ability to induce chondrocyte differentiation of human mesenchymal stem cells were investigated, and cell proliferation changes by needle size differences were determined.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.41 no.2
• /
• pp.51-55
• /
• 2020

The objective of this study was to evaluate the changes in gene expression after incubation of cells with soluble fraction from different silk mat layers. A silk cocoon from Bombyx mori was separated into 4 layers of equal thickness. The layers were numbered from 1 to 4 (from the inner to outer layer). Each silk mat was placed into normal saline and collected soluble fraction. They were administered to RAW264.7 cells, and changes in the expression of genes were evaluated by cDNA microarray analysis. Layer 1 and 4 groups showed significantly higher expression of BMP-2 at 8 h after administration of soluble fraction (P < 0.05). Runx2 expression was significantly higher in Layer 4 group at 8h (P < 0.05). The silk mat from the innermost and outermost portion of the silkworm cocoon showed a significant change in the expression of genes that are associated with osteoinduction such as BMP-2 and runx2.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.34 no.4
• /
• pp.412-418
• /
• 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 the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.26 no.4
• /
• pp.325-336
• /
• 2000

Bone morphogenetic protein-2/4 are members of Transforming Growth Factor-$\beta$(TGF-$\beta$) superfamily and they may induce formation of cartilage and bone in vivo. This study was performed to investigate the cellular target and period of action of BMP-2/4 and understanding of actions of BMP-2/4 at cellular level. The appearance of BMP-2/4 during healing of mandibular and periodontal defect in rat was evaluated immunohistochemically. 40 Sprague-Dawley strain white male rats, each weighing about 300gm were used. Bony defect was performed in the mandible and they were sacrificed at the day of 3rd, 10th, 20th, 30th after operation. The specimens were harvested and examined histologically and immunohistochemically by localization of anti-BMP-2/4. The results were as follows: 1. Woven bone was observed at 10th day and perfect healing of defect with compact bone and periodontal ligment space at 30th day. 2. Osteoprogenitor cells, osteoblastic cells and periosteum were positive reaction to immunohistochemical stain at 10th day. 3. Cells of bone marrow space and surface cells of osteocytes and cementoblasts were positive reaction to immunohistochemical stain at 20th day. 4. Newly formed osteocytes and cementocytes were positive reaction to immunohistochemical stain at 30th day. From the above findings, we could conclude that BMP-2/4 acted significant roles as factors of induction, proliferation and differentiation during bone healing process.

• Journal of Periodontal and Implant Science
• /
• v.41 no.4
• /
• pp.167-175
• /
• 2011

Purpose: Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. Methods: PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 ${\mu}g$/mL ascorbic acid, 10 mM ${\beta}$-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. Results: On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. Conclusions: We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.

• The Journal of Korean Academy of Prosthodontics
• /
• v.50 no.1
• /
• pp.44-52
• /
• 2012

Purpose: In this paper we tried to evaluate the most appropriate surface for rhBMP-2 coating among 4 rough titanium surfaces. Materials and methods: We used machined surface as a control group and anodized, RBM and SLA surfaces as test groups. We coated rhBMP-2 on the 4 surfaces and with uncoated surfaces for each case, we cultured human mesenchymal stem cells on all 8 surfaces. 24 hours after we measured the stem cell' attachment with SEM, and on 3rd, 7th, and 14th days, we checked the cell proliferation and differentiation by using MTT and ALP activity assay. And on the 7th day after the culture, we performed RT-PCR assay to determine whether the expression levels of Type I collagen, osteocalcin, osteopontin were changed. Results: We observed with SEM that 4 rhBMP-2 coated surfaces exhibited wider and tighter cell attachment and more cell process spreading than uncoated surfaces. The anodized rhBMP-2 surface caused robustest effects. In MTT assay we could not find any meaningful difference. In ALP assay there was a significant increase (P<.05) in the ALP activity of anodized rhBMP-2 coated surface compared with that of the control (3rd and 14th days) and with that of the RBM rhBMP-2 coated surface (14th day). In RT-PCR assay there was increased expressions in the anodized rhBMP-2 coated surface for osteocalcin, and osteopontin. Conclusion: We found that the anodized rhBMP-2 coated surface were most prominent stem cell attachment and differentiation in compared to control and Machined rhBMP-2 coated, RBM rhBMP-2 coated surface.