• Title/Summary/Keyword: MTT-test

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Protective Effect of Green Tea Extract and EGCG on Ethanol-induced Cytotoxicity and DNA Damage in NIH/3T3 and HepG2 Cells

  • Kim, Nam Yee;Kim, Hyun Pyo;Heo, Moon Young
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.1-7
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    • 2016
  • In the present study, our aim was to determine whether green tea extract (GTE) and its major constituent, epigallocatechin-3-gallate (EGCG) have a protective effect on ethanol-induced cytotoxicity and DNA damage in NIH/3T3 and HepG2 cells. The cell viability and DNA single strand breaks were examined by MTT assay and alkaline single cell gel electrophoresis (Comet assay), respectively. Ethanol decreased the cell viability and also increased DNA single strand breaks in a concentration-dependent manner. On the other hand, GTE showed the protective effect of cytotoxicity and DNA damage induced by ethanol in both cell lines. GTE and EGCG, were found to possess the anti-oxidative and anti-genotoxic activities by evaluation with DPPH test, LDL oxidation assay, oxidative DNA damage assay and 8OH-2'dG generation test. These results were also verified by the experimental results demonstrating the lower cytotoxicity and genotoxicity of commercial green tea liqueur compared to pure ethanol in same concentration. Thus it is concluded that the supplementation of GTE or EGCG may mitigate the ethanol-induced cytotoxicity and DNA damage.

THE INHIBITORY EFFECT OF TAURINE AND ALENDRONATE ON THE OSTEOCLAST DIFFERENTIATION MEDIATED BY SONICATED EXTRACTS OF PORPHYROMONAS GINGIVALIS IN VITRO. (Porphyromonas gingivalis 분쇄액으로 유도된 파골세포의 분화에 미치는 Taurine과 Alendronate의 효과)

  • Park, Ju-Hyun;Kum, Kee-Yeon;Lee, Jung-Hyun;Yu, Jung-Yun;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.26 no.4
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    • pp.285-295
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    • 2001
  • The objective of this study was to investigate the inhibitory effect of taurine and alendronate on the osteoclast differentiation. Osteoblasts and bone marrow cells from 1-2 day old mouse were co-cultured in 10% fetal bovine serum - minimal essential media (FBS-MEM). Osteoclast differentiation was induced by adding the sonicated extracts of Porphyromonas gingivalis (P.gingivalis). Osteoclasts were identified using tartrate resistant acid phosphotase staining (TRAP). Alendronate of 10$^{-7}$, 10$^{-6}$, 10$^{-5}$M and taurine of 500, 1000, 1500$\mu\textrm{g}$/ml were added respectively. The cytotoxic effects of alendronate and taurine were examined using MTT(3-(4,5-dimethylthiazol -2-yl-2,5-diphenyltetrazo- lium bromide) method. After culturing with the sonicated extracts of P.gingivalis, the amounts of IL-6 in the culture supernatant were measured and compared using the ELISA method. The results were as follows : 1. Osteoclasts were differentiated at the concentration of 0.01~0.1$\mu\textrm{g}$/ml sonicated extracts of P.gingivalis. (P<0.05). 2. Alendronate inhibited osteoclasts differentiation at the concentration of 10$^{-5}$ M when the concentration of sonicated extracts of P.gingivalis was 0.01$\mu\textrm{g}$/ml. 3. Taurine inhibited osteoclasts differentiation at the concentration of 1500$\mu\textrm{g}$/ml when the concentration of sonicated extracts of P.gingivalis 0.01$\mu\textrm{g}$/ml. 4. In cytotoxic test (MTT test), no cytotoxic effect was evident in all concentrations of alendronate and taurine. 5. Taurine (10$^{-5}$M) and alendronate(1500$\mu\textrm{g}$/ml) did not change the amounts of IL-6 induced by sonicated extracts of P.gingivalis significantly.

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Performance Measurement of Split PC Sleepers for Safety in Maintenance Work (분절형 PC침목의 안전성 확보를 위한 성능평가 시험)

  • Eum, Ki-Young;Um, Ju-Hwan;Sin, Seung-Kwon;Lee, Chang-Hun
    • Journal of the Korean Society of Hazard Mitigation
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    • v.6 no.4 s.23
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    • pp.1-6
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    • 2006
  • In terms of the speed-up of the conventional line, the research for a improved turnout was carried out continually around world. A turnout is known that it is the most unstable equipment structurally in the rail system. So, the turnout is needed more stable structure. An accurate assembly of turnout affects traveling qualify of turnout area and running safety. Therefore, a turnout which is pre-assembled at a factory by using the split PC sleeper is transported to construction site recently in advanced countries. In this paper, the performance measurements which estimate the safety of the split PC sleepers which are adopted for the first time to improve performance of turnout are implemented indoors. The split PC sleeper is one of components of the improved turnout. The test is considered of the worst ballast condition under the maintenance work as like STT(Swithc Tie Tamper) or MTT(Multiple Tie Tamper). These experiments results indicate that structural safety for the expansion joint of the split PC sleepers.

Biocompatibility of two experimental scaffolds for regenerative endodontics

  • Leong, Dephne Jack Xin;Setzer, Frank C.;Trope, Martin;Karabucak, Bekir
    • Restorative Dentistry and Endodontics
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    • v.41 no.2
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    • pp.98-105
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    • 2016
  • Objectives: The biocompatibility of two experimental scaffolds for potential use in revascularization or pulp regeneration was evaluated. Materials and Methods: One resilient lyophilized collagen scaffold (COLL), releasing metronidazole and clindamycin, was compared to an experimental injectable poly(lactic-co-glycolic) acid scaffold (PLGA), releasing clindamycin. Human dental pulp stem cells (hDPSCs) were seeded at densities of $1.0{\times}10^4$, $2.5{\times}10^4$, and $5.0{\times}10^4$. The cells were investigated by light microscopy (cell morphology), MTT assay (cell proliferation) and a cytokine (IL-8) ELISA test (biocompatibility). Results: Under microscope, the morphology of cells coincubated for 7 days with the scaffolds appeared healthy with COLL. Cells in contact with PLGA showed signs of degeneration and apoptosis. MTT assay showed that at $5.0{\times}10^4$ hDPSCs, COLL demonstrated significantly higher cell proliferation rates than cells in media only (control, p < 0.01) or cells co-incubated with PLGA (p < 0.01). In ELISA test, no significant differences were observed between cells with media only and COLL at 1, 3, and 6 days. Cells incubated with PLGA expressed significantly higher IL-8 than the control at all time points (p < 0.01) and compared to COLL after 1 and 3 days (p < 0.01). Conclusions: The COLL showed superior biocompatibility and thus may be suitable for endodontic regeneration purposes.

Evaluation of Antiaging Activity on Lycium chinense Miller. Extracts (구기자 추출물의 항노화 활성 평가)

  • Lee, Yong-Hwa;Kim, ․Na-Yeon;Jo, Il-young;Yi, Yong-Sub
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.1
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    • pp.27-33
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    • 2022
  • The purpose of this study was to investigate the functionality of Lycium chinense Miller. as a functional cosmetic material. The total polyphenol content in the extract was 17.3 mg/mL, and as a result of confirming cytotoxicity through MTT assay before the enzyme experiment, it was confirmed that it did not affect cell growth at all concentrations below 1000 ㎍/mL. As a result of conducting DPPH radical scavenging tests as an antioxidant efficacy test, it showed 84.97% scavenging ability at a concentration of 1000 ㎍/mL, and showed very excellent antioxidant efficacy in SOD-like activity test with a result of 80.54% at 1000 ㎍/mL. The measured elastase inhibitory activity, the inhibition rate of 50.93% was observed at 1000 ㎍/mL, indicating that the extract of Lycium chinense Miller. was effective in inhibiting elastase. All these findings suggested that 70% ethanol extract of Lycium chinense Miller. was confirmed as a functional cosmetic material by confirming its antioxidant and anti-wrinkle efficacy.

Synthesis and radiolabeling of PEGylated dendrimer-G2-Gemifloxacin with 99mTc to Biodistribution study in rabbit

  • Mohtavinejad, Naser;Dolatshahi, Shaya;Amanlou, Massoud;Ardestani, Mehdi Shafiee;Asadi, Mehdi;Pormohammad, Ali
    • Advances in nano research
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    • v.10 no.5
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    • pp.461-470
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    • 2021
  • Infection is one of the major mortality causes throughout the globe. Nuclear medicine plays an important role in diagnosis of deep infections such as osteomyelitis, arthritis infection, heart valve and heart prosthesis infections. Techniques such as labeled leukocytes are sensitive and selective for tracking the inflammations but they are not suitable for differentiating infection from inflammation. Anionic linear-globular dendrimer-G2 was synthesized then conjugation to gemifloxacin antibiotic. The structures were identified by FT-IR, 1H-NMR, C-NMR, LC-MS and DLS. The toxicity of gemifloxacin and dendrimer-gemifloxacin complex was compared by MTT test. Dendrimer-G2-gemifloxacin was labeled by Technetium-99m and its in-vitro stability and radiochemical purity were investigated. In-vivo biodistribution and SPECT imaging were studied in a rabbit model. Identify and verify the structure of the each object was confirmed by FT-IR, 1H-NMR, C-NMR and LC-MS, also, the size and charge of this compound were 128 nm and -3/68 mv respectively. MTT test showed less toxicity of the dendrimer-G2-gemifloxacin than free gemifluxacin (P < 0.001). Radiochemical yield was > %98. Human serum stability was 84% up to 24 h. Biodistribution study at 50 min, 24 and 48 h showed that the complex is significantly absorbed by the intestine and accumulation in the lungs and affects them, finally excreted through the kidneys, biodistribution results are consistent with results from full image means of SPECT/CT technique.

The Effect of Treponema Denticola and Treponema Lecithinolyticum on Periodontal Ligament Cells (Treponema Denticola와 Treponema Lecithinolyticum이 치주인대세포에 미치는 영향)

  • Jung, Jung-Hag;Choi, Bong-Kyu;Moon, Ik-Sang;Cho, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.29 no.2
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    • pp.311-326
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    • 1999
  • This study was investigated to observe the effect of Treponema denticola(TDC) and Treponema lecithinolyticum(TLC) on cultured human periodontal ligament cells. Several experiments were performed including MTT test for the inhibition effect of cell proliferation, LDH test for the cytotoxicity , gelatin zymography for the gelatinase activation and observation of cell morphology change using the phase-contrast microscopy. The results were as follows. 1. The effect of concentration on cell proliferation with time showed an inhibitory effect at high concentration $(150{\mu}g/well)$ for TLC and at low concentration( $9.4{\mu}gwell$ ) for TDC. 2. The effect of time on cell proliferation with concentration showed an inhibitory effect at $150{\mu}g/well$ on 2-day incubation for TLC and at $9.4{\mu}g/well$ on 2-day incubation for TDC. 3. The effect of heat-treated TDC and TLC on the inhibition of cell proliferation showed the difference in the heat-treated group compared to the non-heat treated group for TDC, whereas no difference was found for TLC. 4. The morphological changes which were observed from the phase-contrast microscopy showed the difference in the test group compared to the control group. The loss of spindle-like appearance, cell-to-cell detachment and inhibition of cell proliferation were observed. 5. There was no difference of the cytotoxicity effect between the test group and the control group in the LDH test. 6. The active form of progelatinase A with molecular weight 72kDa was activated in both TDC and TLC on the gelatin zymography. Regarding to the above results, TDC and TLC have an effect on periodontal ligament cells by playing an inhibitory role in cell proliferation and appears to activate progelatinase A which degrades type IV collagen.

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AN EXPERIMENTAL STUDY ON FAT CELL VIABLITY ACCORDING TO DIFFERENT HARVESTING TECHNIQUES (지방 채취 방법에 따른 지방 세포의 생존성에 대한 연구)

  • Lee, Won-Deok;Choi, Jin-Young
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.22-29
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    • 2008
  • Purpose: The purpose of this study is to test the efficacy of various methods of fat harvesting in animal model by viability comparison with assay including cell counting, MTT assay, and histologic evaluation. Materials and methods: New Zealand white rabbits experiments were used. Groin fat pads were subjected to different harvest method varying ingredients of solution(Experiment 1: T1 solution= lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 10mgEq/L, Triamcinolone 10mgEq/L; T2 solution=lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 0mgEq/L, Triamcinolone 0mgEq/L) and pressure exerted on harvesting with Luer-Lock syringe connected to suction cannula.(Experiment 2: P1 group=3cc intermittent pressure; P2 group=10cc sustained pressure) Fat cell viability was assessed with cell counting with a hemocytometer, MTT assay, and histologic evaluation. Results: Experiment 1 Cell count: T1=2.4/3.4/4.2, T2=9.6/8.4/7.2($\times10^5$ per mL); MTT assay: T1=0.516/0.41/0.453/0.412/0.421, T2=0.925/0.765/0.54/0.634/0.614 in 21 days(absorbance); Histology: T1 showed elongated and, different in size and shape, and ruptured adipocytes with only a few normal adipocytes whereas T2 showed central core of fat with almost intact fat cells Experiment 2 Cell count: P1=1.2/3.2/4.2, P2=1.2/2.4/3.8($\times10^5$ per mL); MTT assay:P1=0.256/0.245/0.258/0.21/0.264, P2=0.12/0.231/0.245/0.313/0.281 in 21 days(absorbance); Histology: P1 showed somewhat evenly distributed normal-looking fat cells and P2 showed relatively irregular shape of fat cells with small blood vessel amongst adiopocytes. Conclusion: Viability was higher in ‘modified tumescent solution’without sodium bicarbonate and triamcinolone and we also found no significantly different viability between using intermittent pressure and using sustained pressure. But in terms of initial viability of fat cell, we can assume that lower intermittent pressure would make better clinical results.

In vitro cytotoxicity of four kinds orthodontic band cements (수종 치과 교정용 밴드 시멘트의 세포독성에 관한 실험적 연구)

  • Lee, Won-Chul;Park, Soo-Byung
    • The korean journal of orthodontics
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    • v.34 no.4 s.105
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    • pp.351-362
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    • 2004
  • Orthodontic band cements are widely used in the fields of orthodontics, but they are commonly known as cytotoxic material. Within an oral cavity several ions and components are released from orthodontic band cements, thus causing inflammation or injury to the Periodontal tissue. Therefore, it is very important to estimate the biocompatibility of orthodontic band cements. The purpose of this study was to assess the cytotoxic effect of orthodontic band cements to HGF cells. A zinc phosphate cement, a glass ionomer, a resin modified glass ionomer, and compomer were used to evaluate three cytotoxicity assays: cell proliferation assay, MTT assay, and agar ovelay assay The results were as follows: 1. In the cell proliferation assay, Gl>ZPC, RMGI, RMGI24, GI24>compomer24, ZPC24, compomer>metal ring lined up in order of cytotoxicity 2. In the MTT assay, GI>ZPC, RMGI>GI24>ZPC24, compomer, metal ring, RMGI24, compomer24 lined up in order of cytotoxicity. 3. In the agar overlay test, GI>GI24, ZPC, ZPC24, RMGI>RMGI24, compomer, compomer24, metal ring lined up in order of cytotoxicity.

Effect of Acer tegmentosum M. Extracts on Hepatocarcinoma Cell (산겨릅나무 추출물의 간암세포의 증식억제 효과)

  • Kwon, Ha-Na;Bang, Woo-Suk;Kim, Joo-Young;Park, Jyung-Rewng;Jeon, Jeong-Ryae
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.787-790
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    • 2011
  • The objective of this study was to investigate the anticancer effects of Acer tegmentosum M. extracts. HepG2 hepatocarcinoma cells were treated with ethanol, chloroform, ethylacetate, butanol, aqueous fraction and hot water extract. The antiproliferative effect was evaluated by trypan blue exclusion, MTT-based viability assay and morphology. The trypan blue test showed that anticancer effect of the A. tegmentosum M. extracts on HepG2 cells increased gradually in proportion to the increasing concentration of the fractions. The butanol fraction showed the highest anticancer activity against HepG2 cells (p<0.05). The MTT assay indicated that the growth inhibition by the butanol fraction was dose-dependent. These results suggest that A. tegmentosum M. has the potential to inhibit the growth of hepatocarcinoma cells.