• Journal of Korean Medicine for Obesity Research
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• v.18 no.2
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• pp.57-63
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• 2018

Objectives: Metabolic syndrome is defined by a cluster of major cardiovascular risk factors: obesity, insulin resistance, dyslipidemia, and arterial hypertension. Several members of a large family of nonselective cation entry channels, e.g., transient receptor potential vanilloid 4 (TRPV4) channels have been associated with the development of dyslipidemia and hypertension. The purpose of this study was to investigate the effects of Leejung-tang (Lizhong-tang), Rikkunshito, and Bojungikgi-tang (Buzhongyiqi-tang) on TRPV4 channel. Methods: Human embryonic kidney 293 cells stably transfected with the TRPV4 expression vectors were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, $5{\mu}g/mL$ blasticidin, and 0.4 mg/mL zeocin in a humidified 20% $O_2/10%$ $CO_2$ atmosphere at $37^{\circ}C$. Whole-cell patch clamp recordings were obtained using an Axopatch 700B amplifier and pClamp v.10.4 software (Molecular Devices, San Jose, CA, USA), and signals were digitalized at 5 kHz using Digidata 1422A. Results: Leejung-tang and Rikkunshito (10, 30 and 50 mg/mL) had no effects on the TRPV4 whole-cell currents at dose dependent manner. However, Bojungikgi-tang (10, 30, and 50 mg/mL) inhibited the TRPV4 whole-cell currents in a dose dependent manner and the half maximal inhibitory concentration (IC50) of Bojungikgi-tang was 18.2 mg/mL. Conclusions: These results suggest that Bojungikgi-tang plays an important roles to inhibit the TRPV4 channel, suggesting that Bojungikgi-tang is considered one of the candidate agents for the treatment of metabolic syndrome such as dyslipidemia and hypertension.

• Journal of Acupuncture Research
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• v.26 no.5
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• pp.65-75
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• 2009

Objectives : Pyrite is one of the important prescriptions that has been used in oriental medicine for healing of fracture. It is reasonable, therefore, to postulate that native copper affects the process of bone metabolism and bone formation. The purpose of this study is to discover the effect of Pyrite on the healing of tibia fracture. Methods : 1. In vitro test : MG-63 cell in human body and the Pyritum in the ratio of 0.5mg/ml, 1.0mg/ml, 1.5mg/ml, 2.0mg/ml were incubated for 24 hours. After 24 hours, RNA was extracted via trizol reagent (Sigma, USA). In order to understand the activation of osteoblast, the level of OPN mRNA, osteopontin, was measured. 2. In vivo tesgroups normal group, control group and experimental group. Left tibia bones of mice in CON and JT groups were fractured by bone cutters. Pyrite was orally administered to the experimental group. After 14 days, each group's tibia specimen was constructed to observe changes in activation of proinflmmatory cytokines in relation to MIF and IL-6. Also, proliferation of osteoblast and osteopontin were measured via changes in levels of OPN and OPN mRNA. Results : In jn-Titro test, the level of OPN mRNA, osteopontin production was remarkably increased in Pyritum-treated MG-63 cells. In in-vitro test, fractured area in external tibia morphology was increased more in the JT group than that of the CON group. Osteogenesis, endochodrial ossification, and osteoid in fractured area were also increased more in the JT group than that of the CON group. Increase in OPN mRNA, osteopontin level and osteoblast's proliferation were observed. Activation of MIF and IL-6 was confirmed from the fracture region. Conclusions : From the result, development of a new stimulator in healing fracture via pyrite is expected.

• The Journal of Advanced Prosthodontics
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• v.3 no.3
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• pp.152-160
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• 2011

PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

• International Journal of Oral Biology
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• v.42 no.4
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• pp.203-211
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• 2017

The aim of this study was to evaluate the role of demineralized and particulate autogenous tooth, and interleukin-6 in bone regeneration. A demineralized and particulate autogenous tooth was prepared and human osteoblast-like cells (MG63) and human osteosarcoma cells were inoculated into the culture. The rate of cell adhesion, proliferation and mineralization were examined, and the appearance of cellular attachment was observed. An 8 mm critical size defect was created in the cranium of rabbits. Nine rabbits were divided into three groups including: An experimental group A (3 rabbits), in which a demineralised and particulate autogenous tooth was grafted; an experimental group B (3 rabbits), in which a demineralized, particulate autogenous tooth was grafted in addition to interleukin-6 (20 ng/mL); and a control group. The rabbits were sacrificed at 1, 2, 4 and 6 weeks for histopathological examination with H-E and Masson's Trichrome, and immunohistochemistry with osteocalcin. The cell-based assay showed a higher rate of cell adhesion, mineralization and cellular attachment in the experimental group A compared with the control group. The animal study revealed an increased number of osteoclasts, newly formed and mature bones in the experimental group A compared with the control group. Eventually, a higher number of osteoclasts were observed in the experimental group B. However, the emergence of newly formed and mature bone was lower than in the experimental group A. The current results suggest that treatment with demineralized and particulate autogenous tooth and interleukin-6 is not effective in stimulating bone regeneration during the bone grafting procedure.

• Journal of the Korean Ceramic Society
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• v.43 no.8 s.291
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• pp.504-508
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• 2006

Functionally gradient porous $Al_2O_3-(t-ZrO_2)/HAp$ composites consist of 3 layers were fabricated using the multi-pass extrusion process at the various temperatures. The continuous pores were homogeneously formed in the $2^{nd}$ passed samples and their size was about $180{\mu}m$ in diameter. In the porous composites sintered at $1200-1400^{\circ}C$, the relative density and bending strength increased with the sintering temperature. The maximum values of relative density and bending strength in the $2^{nd}$ passed $Al_2O_3-(t-ZrO_2)/HAp$ composites were 62.2% and 107.8 MPa, respectively. In order to investigate the growth behavior of osteogenic cells on the functionally gradient porous $Al_2O_3-(t-ZrO_2)/HAp$ composites, an in vitro test was performed, using human osteoblast-like MG-63 cells. The cells were well attached and grown on the rough surface of the inside of the functionally gradient porous body.

• Food Science and Preservation
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• v.13 no.5
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• pp.649-654
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• 2006

To enhance beneficial effects of citron fruits, anticancer and antioxidant activities of citron fruits extracts were assessed with or without ascorbic acid. Total phenolic acids and flavonoids of fruits peels and flesh extracts were determined. Fruits peels contained more phenolic acids and flavonoids than those detected in flesh extracts. Scavenging activities of 1,1-diphenyl-2-picrylhydrazyl radicals and reducing powers were increased depending on the concentration. The antioxidant activities on oxidation of linoleic acid emulsion incubated at $50^{\circ}C$ were increased but the effect was small to that of butylated hydroxy toluene and ascorbic acid. The anti-tumorigenic effect of these compounds were investigated. They were shown to inhibit the in vitro proliferation of four human tumorigenic cell lines, HT-29, MCF-7, DU-145 and HepG2, in a doso-dependent manner. This study demonstrated that the antioxidant and anticancer activities of citron fruits extracts were derived from their phenols and flavonoids.

• Korean Journal of Materials Research
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• v.17 no.6
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• pp.318-322
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• 2007

Porous ${\beta}-tricalcium$ phosphate $({\beta}-TCP)$ bioceramic was fabricated by pressureless sintering using commercial HAp and different volume percentages of PMMA powders (30-60 vol.%). The range of spherical pore size was about $200-250\;{\mu}m$ in diameter. By increasing the PMMA content, the number of pores and their morphology were dramatically changed as well as decreased the material properties. In case of using 60 vol.% PMMA content, network-type pores were found, due to the necking of the PMMA powders. The values of relative density, elastic modulus, bending strength and hardness of the 60 vol.% PMMA content sample, sintered at $1500^{\circ}C$, were about 46%, 22.2 GPa, 5MPa and 182 Hv respectively. Human osteoblast-like MG-63 cells and osteoclast-like Raw 264.7 cells were well grown and fully covered all of the porous ${\beta}-TCP$ bodies sintered at $1500^{\circ}C$.

• Archives of Pharmacal Research
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• v.26 no.12
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• pp.1029-1035
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• 2003

Osteoporosis is recognized as one of the major hormonal deficiency diseases, especially in menopausal women and the elderly. When estrogen is reduced in the body, local factors such as IL-1 $\beta$ and IL-6, which are known to be related with bone resorption, are increased and promote osteoclastogenesis, which is responsible for bone resorption. In the present study, we investigated whether glucosidic isoflavones (Isocal, PIII) extracted from Sophorae fructus affect the proliferation of osteoblasts and prevent osteoclastogenesis in vitro by attenuating upstream cytokines such as IL-1$\beta$ and IL-6 in a human osteoblastic cell line (MG-63) and in a primary osteoblastic culture from SD rat femurs. Interestingly, IL-1$\beta$ and IL-6 mRNA were significantly suppressed in osteoblast-like cells treated with 17$\beta$-estradiol (E2) and PIII when compared to positive control (SDB), and this suppression was more effective at $10^{-8}$% than at the highest concentration of $10^{-4}$%. In addition, these were confirmed in protein levels using ELISA assay. In the cell line, the cells showed that E2 was the most effective in osteoblastic proliferation over the whole range of concentration ($10^{-4}%-10^{-12}$%), even though PIII also showed the second greatest effectiveness at $10^{-8}$%. Nitric oxide (NO) was significantly (p<0.05) upregulated in PIII and E2 over the concentration range $10^{-6}% to 10^{-8}$% when compared to SDB, without showing any dose dependency. In bone marrow primary culture, we found by TRAP assay that PIII effectively suppressed osteoclastogenesis next to E2 in comparison with SDB and culture media (control). In conclusion, these results suggest that local bone-resorbing cytokines can be regulated by PIII at lower concentrations and that, therefore, PIII may preferentially induce anti-osteoporosis response by attenuating osteoclastic differentiation and by upregulating NO.

• Journal of Periodontal and Implant Science
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• v.33 no.1
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• pp.1-11
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• 2003

Hydroxyapatite(HA) has been extensively used as bone graft materials and tooth implant surface coating materials because of its biocompatibility and osteoconductive properties. However, as HA is intrinsically poor in mechanical properties, zirconia($ZrO_2$) was incorporated with HA as reinforcing phases for improvement of mechanical properties. The purpose of this study was to investigate the biological activities of HA-coated zirconia through the cell proliferation test, measurements of alkaline phosphatase activity, and histologic examination. Four kinds of tested blocks were prepared according to the pore size (300-500${\mu}m$/500-700${\mu}m$) and the porosity (70%/90%). Cell proliferation and alkaline phosphatase activity was measured at 1, 7, 14 days. The number of cells proliferate after 7, 14 days were significantly increased in all groups when compared with that of the first day, but there was no significant difference between the 4 groups at each time period. At the 7 day, alkaline phosphatase activities of cells cultured in 4 groups were higher than that of the first day, but there was no significant difference between the 4 groups at each time period. The human gingival fibroblast and MG 63 cell was used to evaluate the cell cytotoxicity using MTT test. The materials tested in the current study turned out to be non-cytotoxic. In histologic examination(SEM), at 1 day there were many cells attached on the surfaces of all kinds of tested blocks. The number of cells were increased over time. At the 14 day, there were more cells proliferated than 1 day and some of the pores of blocks were partially filled with the proliferated cells. The in vitro response of osteoblast-like cells to the HA-coated zirconia showed comparable effect on transformation comparable to hydroxyapatite.

• Archives of Pharmacal Research
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• v.25 no.5
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• pp.685-690
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• 2002

The mitogen-stimulated serine/threonine kinase $p70^{S6k}$ plays an important role in the progression of cells from $G_0/G$_1$$ to S phase of the cell cycle by translational up-regulation of a family of mRNA transcripts family of mRNA transcripts which contain polypyrimidine tract at their 5 transcriptional start site. Here, we report that $p70^{S6k}$ was constitutively phosphorylated and activated to various degrees in serum-deprived AGS, A2058, HT-1376, MG63, MCF7, MDA-MB-435S, MDA-MB-231 and MB-157. Rapamycin treatment induced a significant dephosphorylation and inactivation of $p70^{S6k}$ in all cancer cell lines, while wortmannin, a specific inhibitor of PI3-K, caused a mild dephosphorylation of $p70^{S6k}$ in AGS, MDA-MB-435S and MB-157. In addition, SQ20006, methylxanthine phosphodiesterase inhibitor, reduced the phosphorylation of $p70^{S6k}$ in all cancer cells tested. Consistent with inhibitory effect of rapamycin on $p70^{S6k}$ activity, rapamycin inhibited [$^3H$]-thymidine incorporation and increased the number of cells at $G_{0}G_{1}$ phase. Furthermore, these inhibitory effects were accompanied by the decrease in growth of cancer cells. Taken together, the results indicate that the antiproliferative activity of rapamycin might be attributed to cell cycle arrest at $G_{0}G_{1}$ phase in human cancer cells through the inhibition of constitutively activated $p70^{S6k}$ of cancer cells and suggest $p70^{S6k}$ as a potential target for therapeutic strategies aimed at preventing or inhibiting tumor growth.