• Title/Summary/Keyword: MTT method.

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The Inhibitory Effect of Ferulic Acid and Related Phenolic Compounds against Cancer Cell Lines (Ferulic Acid와 관련 페놀화합물의 암세포주에 대한 독성억제효과)

  • Han, Du Seok;Chun, Joo Won;Jeon, Sung Woo;Ba다, Seung Hwa
    • YAKHAK HOEJI
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    • v.49 no.5
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    • pp.365-369
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    • 2005
  • The inhibitory effect of ferulic acid and related phenolic compounds on the growth of normal cell lines and can­cer cell line was evaluated by the MTT and XTT methods. Ferulic acid decreased the cell viability of human skin melanoma cells by the MTT method and the cell adhesion activity of human oral epithelioid carcinoma cells by the XTT method. These results suggest that ferulic acid has a potential anticancer activity.

Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method

  • Jang, Sun Mi;Cho, Sin-Yeon;Kim, Eui-Seong;Jung, Il-Young;Lee, Seung Jong
    • Journal of Korean Dental Science
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    • v.9 no.1
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    • pp.1-8
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    • 2016
  • Purpose: The aim of this study was to evaluate the rat periodontal ligament cell viability under $0^{\circ}C/2$ MPa condition up to one week using in vivo 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay. Materials and Methods: As soon as 110 upper molar teeth of rats were extracted, they were stored in Hartman's solution under $0^{\circ}C/2$ MPa condition for 1, 2, 3, 4, and 7 days each. All specimens were treated with in vivo MTT assay and the value of optical density was measured by ELISA reader. These values were statistically analyzed by one-way ANOVA. Result: There was no statistical difference on MTT value between immediate and 1 day storage group. There were statistically significant differences between 1 day and 2 days tsorage, 2 and 3 days storage groups, respectively. Teeth of 34, and 7 days storage groups showed significantly lower MTT valuesc ompared with shorter period storage groups. Conclusion: When the MTT values were substituted in standard curve, 1 day storage group at $0^{\circ}C/2$ MPa condition showed 68% cell viability when compared with immediate group. It dropped to 13% at 2 days, and to less than 5% at 3 days or more.

EVALUATION OF THE VIABILITY OF PERIODONTAL LIGAMENT CELL IN RAT TEETH USING SLOW CRYOPRESERVATION METHOD WITH MAGNETIC FIELD (자기장 저속 냉동보관법을 이용한 쥐 치아 치주인대세포의 활성도 검사)

  • Ahn, Hyun-Jung;Kim, Eui-Seong;Kim, Jin;Kim, Duck-Won;Kim, Ki-Yeol;Lee, Chan-Young;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.33 no.4
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    • pp.332-340
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    • 2008
  • The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4$^{\circ}C$ for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

Changes in Color Response of MTT formazan by Zinc Protoporphyrin (MTT formazan의 발색에 미치는 zinc protoporphyrin의 영향)

  • Park, Kyung-A;Choi, Hyun-A;Kim, Mi-Ri;Choi, Yoo-Mi;Kim, Hyun-Jung;Hong, Jung-Il
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.754-759
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    • 2011
  • Zinc protoporphyrin (ZnPP) is produced endogenously during heme metabolism and treated in cells as a heme oxygenase inhibitor. In the present study, the effects of ZnPP on the color response of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, a commonly-used method for analyzing cell viability, were investigated. ZnPP induced rapid decolorizaion of MTT formazan under light; the degradation rates were 10- and 20- folds faster in the presence of 5 and $10{\mu}M$ ZnPP, respectively. Methylene blue (MB), another type of photosensitizer, also accelerated degradation of formazan under light. Butylated hyroxytoluene did not inhibit ZnPP- or MB-induced formazan degradation. The color degradation of formazan dye was signficantly delayed in the presence of N-acetylcysteine or ${\beta}$-carotene. The present results suggest that certain photosensitizing compounds may affect the color and stability of MTT formazan, which should be carefully considered when conducting the MTT assay.

Screening of Korean Medicinal and Food Plants with Antitumoral Activity (국내 약용 및 식용식물중 항종양활성 식물탐색)

  • Chung, Ill-Min;Kim, Kwang-Ho;Ahn, Joung-Kuk;Ahn, Jong-Seog;Ahn, Soon-Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.1
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    • pp.37-44
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    • 1999
  • This study was conducted to screen antitumoral activity by in vitro bioassay method using 60 Korean medicinal and food plants extracted by 80% EtOH. Antitumor activity test was applied by the PKC(protein kinase C) and antibleb formation, PLC (Phospholipase C), and colorimetric tetrazolium assay (MTT assay) methods. Chenopodjum album and black Glycine max showed high antitumoral activity by 73.5% and 81.0%, respectively, against PKC by bleb-forming assay and PKC enzyme assay on human chronic leukemia K562 cell. Black Glycine max also showed 91.2% antitumoral activity in the PLC method and the lowest $IC_{50}$ $value(4.7{\mu}g/ml)$ by MTT method against P-338 cell line. In the effect of the concentration treatment on antitumoral test, the more concentration indicated the more activity value.

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Effects of Carbofuran and Compensation of Phenobarbital sodium in the NIH 3T3 Fibroblast and Rat Kidney (Carbofuran이 NIH 3T3섬유모세포와 흰쥐의 신장에 미치는 영향과 Phenobarbital sodium의 보상효과에 관한 연구)

  • 한두석;임요섭;한성수
    • Toxicological Research
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    • v.13 no.1_2
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    • pp.87-94
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    • 1997
  • This study was carried out to investigate toxicity of insecticide carbofuran and compensatory effects of phenobarbital sodium (PB) in vivo and in vitro. Sprague Dawley male rats were used as experimental animals and divided into carbofuran only administered group and simultaneous application group of carbofuran and PB. At 30 rain and 1, 3, 6, 12, 24, 48 and 96 hrs after each treatment, the animals were sacrificed by decapitation. Kidney were immediately removed, immersed in fixatives, and processed with routine method for light microscopic study. Paraffin sections were stained with H-E, PAM and PAS. $5.0\times 10^4$ cell/ml of NIH 3T3 fibroblast in each well of 24 multidish were cultured: After 24 hours, the cells were treated with solution of six groups; control group cultured in media only, carbofuran $MTT_50$ or $NR_50$ group cultured in the media containing carbofuran $MTT_50$ or $NR_50$ and four experimental groups cultured in the media containing carbofuran $NR_50$ plus various concentratins of PB. After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours, Tetrazolium MTT (MTT) and NR (neutral red) assay were performed to evaluate the cytotoxicity of cell organelles. Under the light microscope, atrophic change of renal corpuscles were frequently observed in 1 and 2 days after carbofuran treatment. The increase of the mesangium was apparent in 1 and 2 days after carbofuran treatment. Necrotic changes of the epithelium and loss of brush border of proximal tubules were most severe at 2 and 3 days after carbofuran treatment, respectively. In contrast, there were no evidences of the toxic effects on renal tissues at 48hrs in carbofuran-PB treated groups. Carbofuran $MTT_50$ and $NR_50$ were 78$\mu M$, 82.5$\mu M$ respectively. MTT and NR quantities were significantly increased in carbofuran-PB 100$\mu M$ treatment group and carbofuran-PB 100$\mu M$ treatment group. On the basis of these results, it is obvious that PB has compensatory effects against carbofuran toxicity.

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The Flavin-Containing Reductase Domain of Cytochrome P450 BM3 Acts as a Surrogate for Mammalian NADPH-P450 Reductase

  • Park, Seon-Ha;Kang, Ji-Yeon;Kim, Dong-Hyun;Ahn, Taeho;Yun, Chul-Ho
    • Biomolecules & Therapeutics
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    • v.20 no.6
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    • pp.562-568
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    • 2012
  • Cytochrome P450 BM3 (CYP102A1) from Bacillus megaterium is a self-sufficient monooxygenase that consists of a heme domain and FAD/FMN-containing reductase domain (BMR). In this report, the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) by BMR was evaluated as a method for monitoring BMR activity. The electron transfer proceeds from NADPH to BMR and then to BMR substrates, MTT and CTC. MTT and CTC are monotetrazolium salts that form formazans upon reduction. The reduction of MTT and CTC followed classical Michaelis-Menten kinetics ($k_{cat}=4120\;min^{-1}$, $K_m=77{\mu}M$ for MTT and $k_{cat}=6580\;min^{-1}$, $K_m=51{\mu}M$ for CTC). Our continuous assay using MTT and CTC allows the simple, rapid measurement of BMR activity. The BMR was able to metabolize mitomycin C and doxorubicin, which are anticancer drug substrates for CPR, producing the same metabolites as those produced by CPR. Moreover, the BMR was able to interact with CYP1A2 and transfer electrons to promote the oxidation reactions of substrates by CYP1A2 and CYP2E1 in humans. The results of this study suggest the possibility of the utilization of BMR as a surrogate for mammalian CPR.

THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS (압력 저속 냉동 방법의 쥐 치아 치주인대세포 보존 효율 평가)

  • Lee, Young-Eun;Kim, Eui-Seong;Kim, Jin;Han, Seung-Hoon;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.34 no.4
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    • pp.356-363
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    • 2009
  • The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at $4^{\circ}C$for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group. By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.

A Comparative Study of in vitro Methods on the Phototoxicity of Phenothiazines (Phenothiazines의 광독성에 대한 in vitro 실험법의 비교 연구)

  • 김종예;김현진;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.15 no.1_2
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    • pp.13-18
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    • 2000
  • A few in vitro methods were developed to compare the result on the phototoxicity of phenothiazines. By the MTT assay, the Candida test, and the RBC photohemolysis, the phototoxicities of UVA and UVB irradiation were measured. This paper presents the comparisons of methods which are effective to measure the phototoxicities of the chemicals causing phototoxicity and photoallergy. The tested chemicals of phenothiazines include Chlorpromazine, Promethazine, Perphenazine, Chlorprothixene, Trifluoperazine and Thioridazine. Each chemical represented variable results according to the test methods. MTT assay shows the most sensitive method.

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Toxic Effect on Phenolic Compound by Colorimeteric Assay in Normal NIH 3T3 Fibroblasts

  • Jin Byung-Jo;Lee Joo-Hyun;Choi Ki-Wook;Lee Jae-Kyoo;Han Du-Seok
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.263-268
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    • 2004
  • This study was carried out to evaluate the cytotoxic effect of phenolic compound on normal NIH 3T3 fibrolasts. The colorimetric assay for phenol compound, syringic acid was performed by MTT assay or XTT assay. MTT or XTT assays are known as a very sensitive method in measuring the cytotoxic effect of chemical agents in vitro. In the present study, syringic acid on normal Nlli 3T3 fibroblasts did not show any cytotoxicity for MTT assay or XTT assay compared with control after cells were treated with various concentrations of syringic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 3,340.9 μM and 2,462.4 μM of syringic acid, respectively. From the above the results, it is suggested that phenolic compound of syringic acid did not have any cytotoxicity on normal NIH 3T3 fibroblasts.

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