• Title/Summary/Keyword: MTT tetrazolium

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중금속 카드뮴의 세포독성에 관한 연구

  • 정연태;박승택;문연자;한두석;소진탁
    • Toxicological Research
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    • v.9 no.1
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    • pp.45-60
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    • 1993
  • The present study was carried out to evaluate the cytotoxicity of cadmium on cultured rat fibroblasts. The colorimetric assays of neutral red and tetrazolium MTT, the lactatedehydrogenase activity, the amounts of total protein, the rate of DNA synthesis, the amounts of unscheduled DNA synthesis, the frequency of sister chromatid exchange, the releasing rate of intracellular calcium, and light and electron microscopic studies were performed on cultured rat fibroblasts maintained in the media containing various concentrations of cadmium. The results were as follows: The neutral red(NR) and MTT values were decreased dose-dependently by cadmium, and the NR90, NR50, MTT90 and MTT50 values of cadmium were 0.2mM, 21.5mM, 1.0Mm and 60.0Mm, respectively.

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Correlation Between Skin Irritation and Cytotoxicity of Anti-wrinkle Agents (화장품 원료의 피부자극성과 세포독성의 관련성)

  • 이은희;이종권;김용규;박기숙;안광수
    • YAKHAK HOEJI
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    • v.45 no.3
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    • pp.310-319
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    • 2001
  • To compare skin irritation and cytotoxicity of anti-wrinkle agents, we examined skin irritation of six anti-wrinkle agents (ascorbic acid, glycolic acid, all trans-retinoic acid, ginseng extract, retinol, EB) in New Zealand white rabbit. Cytotoxicity of these agents was determined by MTT [tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] at multi-time points in cultured HaCaT cell, a human immortalized keratinocyte cell. We then analyzed correlation between skin irritation and cytotoxicity by spearman's rank correlation analysis. All trans-retinoic acid showed the highest primary irritation index (0.92) in skin irritation test. Being all the six agents not irritant, retinal showed the most cytotoxic agents. The correlation between skin irritation and cytotoxicity ($IC_{50}$/ at different time point was 0.814, 0.757, 0.814 and 0.7 at 3, 24, 48 and 72 h, respectively. We also fecund that IC$_{20}$ and IC$_{80}$ of these agents showed similar correlation with skin irritation. These results therefore demonstrated that there is close correlation between skin irritation and cytotoxicity $IC_{50}$/ value by MTT in HaCaT cell at early time points by anti-wrinkle agents or IC$_{20}$ value. $IC_{50}$/ at earily time point or IC$_{20}$ values may be reliable alternative determinant of skin irritation.n.

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Antitumor Evaluation of Cannabidiol and Its Derivatives by Colorimetric Methods

  • Baek, Seung-Hwa;Shin, Ji-Hee;Chung, Woo-Young;Han, Du-Seok
    • Toxicological Research
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    • v.16 no.2
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    • pp.101-107
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    • 2000
  • Cannabidiol derivatives (1, 2 and 3), 5-fluorouracil (4, 5-FU) and adriamycin (5, AM) were tested for their growth inhibitory effects against human tumor cell lines using two different 3-{4,5-dimeth-ylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. The light microscopic study showed morphological changes of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; MTT assay and STB assay. These results suggest that cannabidiol (1, CBD) retains the most growth-inhibitory activity against human tumor cell lines.

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A STUDY ON THE CYTOTOXICITY OF CYTOSINE ARABINOSIDE AND VINBLASTINE ON CULTURED MOUSE FIBROBLASTS (섬유모세포에 미치는 세포 독성에 관한 연구)

  • Kim, Jae-Min;Kim, Ki-Won;Chung, Yeun-Tai
    • Toxicological Research
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    • v.6 no.1
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    • pp.29-40
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    • 1990
  • Cytotoxic effects of cytosine arabinoside and vinblastine on cultured fibroblasts were determined by colorimetric assays of neutral red (NR) and tetrazolium MTT, and by mutagenicity tests . Cytosine arabinoside and vinblastine were highly toxic by showing that concentrations of NR-50 and MTT-50 of two drugs were lower than 100 ${\mu}$M. At mid-point cytotoxicityvalue of two drugs, frequencies of micronuclei and SCEs were very high and chromosome showed structural abnormalities. The sizes of micronuclei formed by vinblastine were larger than those induced by cytosine arabinoside. These results suggest that cytosine arabinoside and vinblastine have highy mutagenic and severe cytotoxic effects on the cultured mouse fibroblasts.

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Effects of Fructus Schisandrae Water Extract on Cultured Mouse Myocardial Cells Induced by Xanthine Oxidase/Hypoxanthine (오미자 추출물이 산소지유기에 의하여 손상된 생쥐의 배양 심근세포에 미치는 영향)

  • 주은정
    • Journal of Nutrition and Health
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    • v.33 no.7
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    • pp.739-744
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    • 2000
  • The purpose of this study was to elucidate protective effect of Fructus Schsandrae(FS) water extract against xanthine oxidase/hypoxanthine(XO/HX)-induced cardiotoxicity in myocardial cells this experiment was performed. Cardiotoxicity of XO/HX was examined by MTT(MTT [3-(4,5-dimethylthiazol-2-yl)-2.5,-diphenyl tetrazolium bromide) assay. XO/HX induced the decrease of cell viability. Also XO/HX induced the increase of LDH activity and the decrease of beating rate on cultured myocardial cells in a dose-dependent manner. To investigate cardioprotective effect of FS water extract cultures were preincubated with FS water extract for 3 hours. Cultures were then exposed to XO/HX for 72 hours. FS water extract have an efficacy in decreaasing LDH activity and increasing heart beating rate on cultured myocardial cells damaged by XO/HX. From the results it is suggested that XO/HX may show toxic effect in cultured myocardial cells derived from neonatal mouse and FS water extract is effective in the prevention of XO/HX-induced cardiotoxicity.

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Decreased Effectiveness on Cytotoxicity of Metal-Metal and Metal-Chelator Combinations (중금속 상호간의 작용 및 착화제에 의한 세포독성의 억제효과)

  • Kim, Jai-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.1 no.1
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    • pp.115-118
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    • 1996
  • The study on the cytotoxicities of heavy metals, metal-metal and metal-chelator combinations was carried out to evaluate the cytotoxic effect of those on mouse L929 fibroblasts. The colorimetric assays (NR and MTT) were conveniently carried out in 96-well microtiter plates. The rank order was Cd > Zn Ni > Cr(III) for the heavy metals tested. Examination of the effect of metal-metal interaction on cytotoxicity showed a moderate reduction of cadmium toxicity by zinc. The colorimetric assays were also effectively used to investigate the effect of the chelators, ethylenediamine tetra acetic acid (EDTA) and chitosan. Reduction of heavy metal toxixity by chelator was efficient.

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In Vitro Skin Irritation Test of Anti-Inflammatory Drugs (소염진통제 약물에 대한 In vitro 피부자극 시험연구)

  • Lee, Jong-Kwon;Kim, Dai-Byung;Lee, Eun-Hee;Lee, Sun-Hee;Ryu, Seung-Rel;Choi, Ki-Hwan;Kim, Yoon-Jeong;Kim, Pu-Young
    • Toxicological Research
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    • v.14 no.3
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    • pp.315-320
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    • 1998
  • In vitro skin iritation of anti-inflammatory drugs was investigated in terms of the cytotoxicity method to human skin fibroblast cells. Five anti-inflammatory drugs (Diclofenac, Naproxen, Meclofenamic acid, Ibuprofen and Fnoprofen) which are commercially available as oral preparations or injections were tested. The cytotoxicity of 5 chemicals was evaluated by using MTT[tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. NRU (neutral red uptake) assay and Alamar Blue assay after fibroblast cells had been exposed to the chemicals for 24 hours or 489 hours. The $IC_{50}$ values of the chemicals showed the comparative strength of cytotoxicity as following order of Meclofenamic acid>Diclofenac>Fenoprofen>Ibuprofen>Naproxen. The values of $IC_{50}$ determined by Alamar Blue assay were lower than those of MTT and NRU assay. These data suggest Alamar Blue assay can be useful method for assessing in vitro skin irritation potential of anti-inflammatory drugs.

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AN EXPERIMENTAL STUDY ON THE CHANGE OF THE RADIOSENSITIVITY OF SEVERAL TUMOR CELL LINES AND PRIMARY CULTURED GINGIVAL FIBROBLAST (수종의 암세포주와 치은섬유아세포에서 방사선의 양과 분할조사에 따른 세포활성도와 독성의 변화에 관한 연구)

  • Lee Sam-Sun;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.107-122
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    • 1997
  • Radiation sensitivity data was generated for two human cancer cell lines(KB, RPMI 2650) and human primary gingival fibroblast was tested three times using a viable cell number counting with a hemocytometer, MTT(3-[4,5-Dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide) assay, and LDH(Lactate dehydrogenase) assay. Single irradiation of 2, 4, 6, 10, 15, 20Gy were applied to the tumor cell lines and the primary cultured gingival fibroblast The two fractions of 4Gy and 10Gy were seperated with a 4 hour time interval. The irradiation was done with 241.5cGy/min dose rate using /sup 137/Cs MK cell irradiator at room temperature. The obtained results were as followed : 1. There was significantly different viable cell numbers as the amount of radiation dose on the tested cells were cell number counted with a hemocytometer. In fractions, there were more viable cells remaining. 2. Phase-contrast microscopically, radiation-induced morphologic changes were pronounced on the tumor cells, however, almost no differences on the gingival fibroblast. 3. There was significantly different absorbance at 2Gy on RPMI 2600, 4Gy on KB and GF in MTT assay. In fractions, the absorbance was significantly higher on KB. 4. The level of extracellular LDH activity in the experimental group was significantly higher in the 2-4Gy than the control group. 5. The total level of extracellular and intracellular LDH activity was decreased as increased amounts of radiation dose was applied.

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Development of Antitioxic Agents from Korean Medicinal Plants. Part 5. Antitoxic Effects of Binding of Caffeic acid and Cadmium on Cultured Rat Neuroglial Cells (한국산 생약으로부터 해독물질의 개발(제5보) 배양 백서 신경아교세포에서 Caffeic acid와 카드뮴의 결합에 의한 해독효과)

  • Baek, Seung-Hwa;Lee, Hong;Pae, Hyun-Ock;Kim, Young-Ok;Kwak, Jung-Suk;Yoo, Hyeon-Gyeong;Han, Du-Seok
    • Toxicological Research
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    • v.11 no.2
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    • pp.241-246
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    • 1995
  • This study was carried out to develop the antitoxic compound about cytotoxicity of cadmium on cultured rat neuroglial cells. These cells divided into 3 groups; control group (medium only) or $MTT_{50}$ group (neuroglial cell, $61{\mu}M$ cadmium) and experimental group ($61{\mu}M$ caffeic acid). Neutral red (NR) and tetrazolium MTT of the colorimetric assay were performed to evaluate the cytotoxicity of cell organelles. The light microscopic study was carried out to morphological changes of cultured rat neuroglical cells. The results indicated that caffeic acid showed detoxification effect on cytotoxicity of cadmium in $61{\mu}M$ concentration. According to the spectroscopic study of 1:1 complex of cadmium and caffeic acid, it showed that this formation of complex eliminated cadmium from cultured rat neurogllal cells.

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Anti-proliferating Effects of Some Plants on the Hepatoma Cell (수종 식물들의 Hepatoma cell 증식 저해작용)

  • Chung, Yong Za;Lee, Un Joo
    • Korean Journal of Clinical Pharmacy
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    • v.9 no.2
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    • pp.103-108
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    • 1999
  • The anti-proliferating effects of some plants on hepatoma cell lines were studied by the 3-[4,5-dim-ethylthiazol-2yl]-2,5-diphenyl-tetrazolium bromide (MTT assay), to investigate the anticancer effect with some plants around here. As the result, we saw that the anti-prolferating effect to the plants. Among the plants, Equisetum arvense L. and Lactuca dentata Makino. var, flaviflora Makino of them relatively showed a good ant-proliferating effect. Capsicum annuum L. var. angulosum Mill (Leaf) was the best among them. We also examined morphological changes on the hepatoma cells in this process. In case of Capxicum annuum L. var. angulosum Mill, the tells become vague after 2 days, and then destroyed faster than others. We can fee also the condensated chromosome on the treated cells with Capxicum annuum L. var. angulosum Mill. And we also observed condensation through using a fluorescent microscope by PI staining, and observed DNA fragmentation.

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