• The korean journal of orthodontics
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• v.32 no.3 s.92
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• pp.227-234
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• 2002

In order to investigate the action mechanism of protein kinase C on $K^+$ channel in osteoblastic cell, effects of phorbol 12, 13-dibutyrate on human osteoblast-like cells (G292) were studied by patch clamp technique with cell-attacked configuration. 111 this experiment, 45pS ion channel was dominant in G292 cell line according to their approximate conductances in symmetrical 140mM KCl saline at holding potential of 60mV. In torrent-voltage relationship, reversal potential was 5.5mV at the condition of potassium enriched saline in the pipette and -27 mV at the condition of standard extracellular saline In the pipette. Phorbol 12, 13-dibutyrate 10nM increased the open probability of 45pS channel and staurosporine, an inhibitor of protein kinase C, suppressed this effect. Phorbol 12,13-dibutyrate moved the reversal potential of 45pS channel to more negative potential and increased the single channel current at the same membrame potential. In order to check the activation of protein kinase C in G292 cell by phorbol 12,13-dibutyrate, western blot of protein kinase C was performed. Phorbol 12,13-dibutyrate $0.1{\mu}M$ translocated protein kinase C from cellular compartment to membrane compartment of the cell. These findings suggest that phorbol 12,13-dibutyrate, one of phorbol esters, activate 45pS channel In G292 cell and affect cell membrane potential, that regulate cellular function.

• Proceedings of the Korean Institute of Surface Engineering Conference
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• 2017.05a
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• pp.79-79
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• 2017

Metallic biomaterials have been mainly used for the fabrication of medical devices for the replacement of hard tissue such as artificial hip joints, bone plates, and dental implants. Because they are very reliable on the viewpoint of mechanical performance. This trend is expected to continue. Especially, Ti and Ti alloys are bioinert. So, they do not chemically bond to the bone, whereas they physically bond with bone tissue. For their poor surface biocompatibility, the surface of Ti alloys has to be modified to improve the surface osteoinductivity. Recently, ceramic-like coatings on titanium, produced by plasma electrolytic oxidation (PEO), have been developed with calciumand phosphorus-enriched surfaces. A lso included the influences of coatings, which can accelerate healing and cell integration, as well as improve tribological properties. However, the adhesions of these coatings to the Ti surface need to be improved for clinical use. Particularly Silicon (Si) has been found to be essential for normal bone, cartilage growth and development. This hydroxyapatite, modified with the inclusion of small concentrations of silicon has been demonstrating to improve the osteoblast proliferation and the bone extracellular matrix production. Strontium-containing hydroxyapatite (Sr-HA) was designed as a filling material to improve the biocompatibility of bone cement. In vitro, the presence of strontium in the coating enhances osteoblast activity and differentiation, whereas it inhibits osteoclast production and proliferation. The objective of this work was to study Morphology of bone-like apatite formation on Sr and Si-doped hydroxyapatite surface of Ti-6Al-4V alloy after plasma electrolytic oxidation. Anodized alloys was prepared at 270V~300V voltages with various concentrations of Si and Sr ions. Bone-like apatite formation was carried out in SBF solution. The morphology of PEO, phase and composition of oxide surface of Ti-6Al-4V alloys were examined by FE-SEM, EDS, and XRD.

• Journal of Acupuncture Research
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• v.29 no.6
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• pp.11-21
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• 2012

Objectives : BHH10 is traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify BHH10 extract induces osteogenic activity in human osteoblast-like MC3T3-E1 cells. Methods : MC3T3-E1, pre-osteoblast cell line, were treated with BHH10 of various concentrations($0.1{\mu}g/mL$, $1{\mu}g/mL$, $10{\mu}g/mL$). And then, the effect of BHH10 on osteoblast differentiation was examined by alkaline phosphatase(ALP) activity, von Kossa staining and RT-PCR for osteoblast differentiation markers such as osteocalcin(OCN), osteopontin(OPN). Results : BHH10 had dose-dependent effect on the viability of osteoblastic cells, and dose-dependently increased alkaline phosphatase(ALP) activity. BHH10 markedly increased mRNA expression for OCN, OPN in MC3T3-E1 cells. Also, BHH10 significantly induced mineralization in the culture of MC3T3-E1 cells. Conclusions : In conclusion, these results propose that BHH10 can play an important role in osteoblastic bone formation, osteogenesis, and may possibly lead to the development of bone-forming drugs.

• The Journal of the Korean dental association
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• v.46 no.9
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• pp.563-573
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• 2008

Purpose : The purpose of this study is to investigate the effects of Simvastain, which is HMG-CoA reductase inhibitor, on proliferation and differentiation of osteoblast. Materials & Methods : Twenty-four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x $10^4$ cells per plate. Each plates were incubated with 5% $CO^2$incubator $37^{\circ}C$. Starting from 2 days after incubation, cell culture medias were replaced with Osteogenesis induction media every 2 days, for 12 days. In some plates, 0.01, 0.1, 1, 10, $100\muM$ of Simvastatin were added with Osteogenesis induction media, and classified as "test group". Those not added with Simvastatin were classified as "control group". Results : 1. When Alrizarin Red staining was observed with naked eye, control group showed normal deep red color, but test group show rapid decrease of red color as Simvastatin concentration increased more than $0.1\muM$. 2, When observed with microscope, compared to control group, amount of osteo matrix stained with Alrizarin Red decreased rapidly in Simvastatin concentration more than $0.1\muM$. 3. In optical density analysis, regarding control group as a basis, mineral deposition decreased rapidly when Simvastatin concentration increased more than $0.1\muM$. 4. In flow cytometry analysis, survival rate of UMR-106 cell showed no changes in both control group and test group. Conclusion : From the above results, we were able to identify that Simvastatin inhibited osteogenesis without effecting survival or cell number of osteoblasts.

• YAKHAK HOEJI
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• v.58 no.5
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• pp.307-313
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• 2014

Rumex crispus (curled dock), which is a perennial wild plant, has long been used as a laxative, astringent, and medicine to treat blood and skin diseases. We recently reported that the roots of R. crispus are an effective nutraceutical for bone. This study prepared ethanol extracts of the leaves and roots of R. crispus, and analyzed the major constituents using liquid chromatography and mass spectrometry. In addition, their effects on the proliferation and differentiation of human osteoblast-like MG-63 cells, such as cell viability, alkaline phosphatase (ALP) activity, collagen content, and mineralization, were compared. The chromatograms of the chemical constituents of the two extracts exhibited quite different profiles: quercetin and quercitrin were identified as major peaks in the leaf extract, whereas cinnamtannin B1 and procyanidin isomers were the major peaks for the root extract. Neither extract was cytotoxic at concentrations of < $25{\mu}g/ml$. ALP activity and collagen synthesis-which are markers of the early stage of osteogenesis-in MG-63 cells were significantly increased upon the addition of the root extract compared with the addition of the leaf extract. In contrast, the leaf extract had a more stimulatory effect on mineralization-which is marker of the late stage of osteogenesis-in MG-63 cells than did the root extract. In conclusion, extracts of both leaves and roots of R. crispus stimulated the bone-forming activity of osteoblasts; in particular, the root extract was more effective in the early stage of osteoblast differentiation, while the leaf extract was more effective in the late stage. This difference in anabolic activity may be due to differences in the constituents of the leaves and roots. The leaves and roots of R. crispus appear to complement each other as stimulators of bone formation.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.1-12
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• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• Archives of Pharmacal Research
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• v.26 no.11
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• pp.917-924
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• 2003

This research aims to test a new drug candidate based on a traditional medicinal herb, F1, an herbal extract obtained from Astragalus membranaceus and its main ingredient, 1-monolinolein that may have fewer side effects and less uterine hypertrophy. In vitro experiments, human osteoblast-like cell lines, MG-63 and Saos-2, were analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and an alkaline phosphatase (ALP) assays. Mouse osteoclasts were induced through a calcium-deficient diet and inhibition effects were measured. In vivo experiments were done using ovariectomized (OVX) rats for 9 weeks. At necropsy, uterus weights were measured, trabecular bone area (TBA) of tibia and lumbar vertebra were measured bone histomorphology. In results, cell proliferation and ALP activity in Saos-2 by ether F1 or 1-monolinolein did not increased significantly compared to the control. The F1 inhibited osteoclast development ($IC_{25}=3.37{\times}10^{-5}$mg/mL) less than 17$\beta$-estradiol. The OVX rats administered F1 (2 mg/kg/day and 10 mg/kg/day) showed an increase in TBA of the tibia significantly (136.3${\pm}4.2% and 138.5{\pm}$10.3% of control). In conclusions, the herbal extract, F1 inhibited tibia and lumbar bone loss and did not cause uterine hypertrophy. However, 1-monolinolein, the main ingredient of the herbal extract, did not inhibit bone loss.

• Proceedings of the Materials Research Society of Korea Conference
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• 2011.05a
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• pp.54.2-54.2
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• 2011

DTBP (dimethyl 3,3`-dithiobispropionimidate) was applied to collagen and gelatin coating on BCP granules and a crosslinking agent. The DTBP crosslinking was done for decreasing the solubility of the coating and hence increasing the stability. The nanostructure of collagen and gelatin coating surfaces were observed by SEM technique. Based on the DSC thermograms and FT-IR spectrums, the crosslinkings were confirmed between collagen molecules and gelatin molecules. The compressive strength was measured before crosslinking and after that. In-vitro study was carried out by measuring cell viability and observing cell morphology after DTBP crosslinking. Moreover, the proliferation ability of MG-63 osteoblast-like cells on the crosslinked BCP granules was evaluated by Western blot assay. The BCP granules were implanted into rabbit femur for 4 weeks and 12 weeks. The bone tissue formation was analyzed with micro-computed tomography (micro-CT) and histological analysis was also carried out by hematoxylin and eosin (H&E) staining for visualization of cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.19 no.4
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• pp.395-398
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• 1997

Ipriflavone(IP)은 골흡수 억제효과에 골형성 촉진효과를 지니는 약물로 보고되어 왔다. 이러한 IP의 특성때문에 골의 치유를 촉진시키기 위한 약물로서 구강외과 영역에서 쓰일 수도 있으리라 생각되었다. 이에 저자는 그동안 보고되어온 IP의 골 형성 촉진효과가 실제로 나타나는지를 확인하고 또한 어떤 농도에서 나타나는지를 알아보기위해 IP를 서로다른 농도로 하여 골세포주(MC3T3-El cell line)의 배지에 넣은후, 골 형성의 지표로 쓰일수있는 collagen합성정도를 보고자 하였으며 이 자료를 앞으로서 in vivo 동물실험 연구의 기초자료로 사용코자 하였다. 본 연구에서 IP의 골형성 촉진효과를 collagen 합성정도를 측정을 통해 확인하였고, 특히 IP이 $10^{-7}M$농도일때 현저한 collagen합성의 증가를 관찰 하였으며 앞으로의 동물실험등을 통해 구강외과 영역에서의 사용가능성에 대해 좀더 연구해 보고자 한다.

• IMMUNE NETWORK
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• v.16 no.1
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• pp.26-32
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• 2016

Aortic valve stenosis is a heart disease prevalent in the elderly characterized by valvular calcification, fibrosis, and inflammation, but its exact pathogenesis remains unclear. Previously, aortic valve stenosis was thought to be caused by chronic passive and degenerative changes associated with aging. However, recent studies have demonstrated that atherosclerotic processes and inflammation can induce valvular calcification and bone deposition, leading to valvular stenosis. In particular, the most abundant cell type in cardiac valves, valvular interstitial cells, can differentiate into myofibroblasts and osteoblast-like cells, leading to valvular calcification and stenosis. Differentiation of valvular interstitial cells can be trigged by inflammatory stimuli from several immune cell types, including macrophages, dendritic cells, T cells, B cells, and mast cells. This review indicates that crosstalk between immune cells and valvular interstitial cells plays an important role in the development of aortic valve stenosis.