• Imaging Science in Dentistry
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• v.39 no.3
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• pp.121-132
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• 2009

Purpose : To investigate the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of bone sialoprotein (BSP) and osteocalcin (OC) during the differentiation in irradiated MC3T3-E1 osteoblastic cells. Materials and Methods : When MC3T3-E1 osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or $10{\mu}M$ QCT, and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the synthesis of type I collagen, and expression of BSP and OC. Results : The synthesis of type I collagen in cells exposed to 2 Gy of radiation in the presence of 2-DG or QCT showed no significant difference compared with the control group within 15 days post-irradiation. When the cells were irradiated with 8 Gy, 2-DG facilitated the irradiation mediated decrease of type I collagen synthesis, whereas such decrease was inhibited by treating with QCT. During MC3T3-E1 osteoblastic cell differentiation, the mRNA expression of BSP and OC showed the peak value at 14 days and 21 days, respectively. 2-DG or QCT treatment alone decreased the level of BSP mRNA, but increased the OC mRNA level only at early time of differentiation (day 7). In the cells irradiated with 2, 4, 8 Gy, the mRNA expression of BSP and OC decreased at 7 days after the irradiation. The cells were treated with various dose of radiation in the presence of 2-DG or QCT, the mRNA level of both BSP and OC increased although this increase was observed at low dose of radiation (2 Gy) and at the early stage of differentiation. However, when the cells were exposed to 4, 6, or 8 Gy, the increase of BSP and OC mRNAs was detected only in cells co-incubated with QCT. Conclusion : This study demonstrates that 2-DG and QCT affect differently the expression of bone formation related factors, type I collagen, BSP, and OC in the irradiated MC3T3-E1 osteoblasic cells, according to the dose of radiation and the times of differentiation. Overall, the present findings suggest that 2-DG and QCT could have the regulatory roles as radiation-sensitizer and -protector, respectively.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.2
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• pp.1-18
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• 2009

Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.

• Journal of Korea Foundry Society
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• v.20 no.4
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• pp.260-268
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• 2000

응고 및 마모거동을 연구하기 위하여 Cr, V, Mo 및 W의 조성이 다른 3종류의 백주철을 주조하였다. 고주파유도용해로에서 장입시켜 1873K까지 승온시켜 장입원료들을 모두 용해시킨 후 1823K에서 Y블럭주형에 주입하였다. 얻어진 3종류의 백주철조성은 다음과 같다: Fe-3%C-10%Cr-5%Mo-5%W(합금1), Fe-3%C-10%V-5%Mo-5%W(합금2) 및 Fe-3%C-17%Cr-3%V(합금 3)응고과정을 추적하기 위하여 각 시편으로부터 50g을 채취한 후 알루미나도가니에 넣고 실리콘카바이드로를 사용하여 1723K의 알콘분위기하에서 재용해를 시켰다. 용금을 10K/분의 속도로 냉각시키면서 열분석을 행하였으며 도중 몇 차례 소입 실험을 병행하였다.합금1의 경우 초정오스테나이트, (오스테나이트+$M_7C_3$)공정, (오스테나이트+$M_6C$)공정, 합금2의 경우 초정 MC, 초정오스테나이트, (오스테나이트+MC)공정 , (오스테나이트+$M_2C$)공정, 합금3의 경우 초정 $M7C_3$와 (오스테나이트+$M_7C_3$)공정으로 구성되어 있었다. 주방상태, 균질화열처러상태, 공냉상태, 템퍼링상태에서 내마모시험을 행하였다. 먼저 주방상태시 편을 진공분위기하에서 1223K에서 5시간동안 균질화열처리를 행한후 로냉을 시켰다. 다시 이 시편을 1323K에서 2시간 유지후 강제공냉을 시켰으며 강제공냉된 시편을 573K에서 3시간동안 템퍼링처리를 하였다. 내마모시험은 120 mesh마모지에 10N의 하중을 가하여 실시하였다. 각 사이클마다 무게감소를 측정하였으며 8번 반복실험을 하였다. 마모량은 균질화열처리시편, 주방상태시편, 템퍼링시편, 공냉시편의 순으로 감소하였다. 합금2가 마모량이 가장 적었으며 합금3이 가장 많았다. 합금2의 마모량이 가장 적은 이유는 조직이 초정 MC, 공정 MC 및 공정 M2C로 구성되어 있기 때문으로 사료된다.주방상태에서 기지조직은 퍼얼라이트이었으나 열처리를 통하여 마르텐사이트, 템퍼드마르텐사이트, 잔류오스테나이트로 변태하였다. 이상의 결과를 종합해 보면 MC가 내마모성에 가장 기여하는 조직으로 판단되어진다.

• The korean journal of orthodontics
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• v.28 no.1 s.66
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• pp.155-163
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• 1998

To evaluate the effect of a static magnetic field on the bone producing potential of MC3T3-E1 cells, the alkaline phosphatase activity was measured after the cells having been cultured under 76.4mT static magnetic field using a $SmCo_5$ magnets for 5days, 7days, 11days, 15days and 21days for each cell culture group. Also, the amount of bone nodule stained with Alizarin red S was observed. The results were as follows . The alkaline phosphatase activity of the 7, 11, and 15 days group among the experimental groups was decreased as compared with the control groups, and the decrease of alkaline phosphatase activity in the 11 days group was the most evident among them. . Any stained bone nodules of both groups had not been observed until the 11th day. The stained bone nodules in the control groups were found on the 15th day, but not in the experimental groups. The stained bone nodules were observed in both groups on the 21st day, but the control groups have more bone nodules than the experimental groups.

• The Journal of Internal Korean Medicine
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• v.36 no.4
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• pp.518-526
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• 2015

Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.

• Journal of Life Science
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• v.27 no.5
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• pp.501-508
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• 2017

Bone morphogenetic proteins (BMPs) are multifunctional cytokines that play important roles in a variety of cellular functions. Among BMP family members, BMP2 efficiently promotes osteoblast differentiation through Smad-mediated runt-related transcription factor 2 (Runx2) expression. Several recent studies suggest that BMPs are associated with clock genes, in particular Bmal1. Bmal1 protein heterodimerizes with Clock protein and then induces period 1 (Per1) expression. However, the role of Per1 on osteoblast differentiation remains unclear. In this study, we investigated whether Per1 is involved in osteoblast differentiation. MC3T3-E1 cells were treated with BMP2 for induction of osteoblastic differentiation. Osteogenic maker gene and Per1 mRNA expression were measured using real-time PCR. Interestingly, BMP2 treatment induced Per1 mRNA expression in MC3T3-E1 cells. To further investigate the function of Per1 on osteoblast differentiation, MC3T3-E1 cells were transiently transfected with pCMV-Per1. Per1 overexpression increased Runx2 mRNA and protein levels. Also, mRNA expression and promoter activity of osteocalcin were upregulated by Per1 overexpression. To investigate the effect of interaction between Per1 and osteogenic condition, MC3T3-E1 cells were cultured in osteogenic medium containing ascorbic acid and ${\beta}$-glycerophosphate. Osteogenic medium-induced ALP staining level and mineralization were synergistically increased by overexpression of Per1. Taken together, these results demonstrate that Per1 is a positive regulator of osteoblast differentiation.

• Journal of dental hygiene science
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• v.20 no.4
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• pp.221-229
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• 2020

Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.807-811
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• 2007

Diabetes is marked by high glucose levels and is associated with decreased bone mass and increased fracture rates. To determine if [6]-gingerol could influence osteoblast dysfunction induced by 2-deoxy-D-ribose (dRib), osteoblastic MC3T3-E1 cells was treated with dRib and [6]-gingerol and markers of osteoblast function and oxidized protein were examined. [6]-Gingerol ($10^{-7}\;M$) significantly increased the growth of MC3T3-E1 cells in the presence of 30 mM dRib (p<0.05). [6]-Gingerol ($10^{-7}\;M$) caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and osteocalcin secretion in the cells. We then examined the effect of [6]-gingerol on the production of osteoprotegerin and protein carbonyl in osteoblasts. Treatment with [6]-gingerol ($10^{-9}$ and $10^{-7}\;M$) increased osteoprotegerin secretion in osteoblastic cells. Moreover, [6]-gingerol ($10^{-9}$ and $10^{-7}\;M$) decreased protein carbonyl contents of osteoblastic MC3T3-E1 cells in the presence of 30 mM dRib. Taken together, these results demonstrate that [6]-gingerol inhibits dRib-induced damage and may be useful in the treatment of diabetes related bone diseases.

• International Journal of Oral Biology
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• v.39 no.3
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• pp.145-151
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• 2014

During bone remodeling, there is requirement of differentiation of osteoblastic cells. Previously, we identified proteins differentially expressed in soft tissue during bone healing. Of these proteins, we focused the effect of LTF on differentiation of osteoblast. In order to analyze the osteogenic ability of LTF, we treated conditioned media collected from human LTF-stably transfected HEK293T cells into osteoblastic MC3T3-E1. The results showed that the activity and expression of alkaline phosphatase were increased in MC3T3-E1 cells treated with conditioned media containing LTF in dose- and time-dependent manner. At the same time, we observed the significant increase of the expression of osteoblastic genes, such as ALP, BSP, COL1A1, and OCN, and along with matrix mineralization genes, such as DMP1 and DMP2, in LTF conditioned media-treated groups. Moreover, the result of treating recombinant human LTF directly into osteoblastic MC3T3-E1 showed the same pattern of treating conditioned media containing LTF. Our study demonstrated that LTF constitutively enhances osteoblastic differentiation via induction of osteoblastic genes and activation of matrix mineralization in MC3T3-E1 cells.

• The Journal of the Petrological Society of Korea
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• v.10 no.3
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• pp.157-171
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• 2001

This study investigated the effects of Pb/Tl ratio, Pb concentration and concomitant matrix elements on the measurement of Pb isotope ratios using multi-collector ICP/MS (AXIOM MC model). Accuracy and reproducibility of Pb isotope ratios in NBS 981 solution were estimated for 42 data measured from March to August 2001. Pb isotopes measured in rocks, bronzes and sediments were compared to data measured by TIMS. Reproducibilities for $^{206}Pb/^{204}Pb,\; ^{207}Pb/^{204}Pb,\;and\;^{208}Pb/^{204}Pb$ ratio were about 500 ppm (2sd) and for $^{207}Pb/^{206}Pb$\;and\;^{208}Pb/^{206}Pb$ were 100~200 ppm for 200 ng of Pb in NBS 981 solution. The optimum conditions for the analysis of Pb isotope ratios with AXIOM MC for best accuracy and reproducibility were defined as follows; 1) Pb/Tl ratio is about 10 2) Pb concentration is about 100 ng/ml 3) correction for mass discrimination is performed by exponential law using 2.3887 of $^{205}Tl/^{203}Tl$ and Pb mass fractionation factor empirically obtained from $ln(^{208}Pb/^{206}Pb)-ln(^{205}Tl/^{203}Tl)$ relationship. The sample data measured with MC/ICP/MS for acid-digested and chemically separated rock samples, and acid-digested bronze samples and sediment samples coincide with those of TIMS within analytical errors. Therefore, MC/ICP/MS is a rapid analytical technique for Pb isotope ratios with the similar precision compared with TIMS.