• Title/Summary/Keyword: Colorimetric Assay

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A Study on the Cytotoxic Effects of Several Plant Extracts on the Cell viability and Cell Adhesion Activity in Cultured NIH3T3 Fibroblast (몇 가지 식물추출물이 배양 NIH3T3 섬유모세포의 세포생존율과 세포부착률에 미치는 세포독성에 관한 연구)

  • Rim, Yo-Sup;Song, Won-Seob;Seo, Young-Mi;Park, Seung-Taeck;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.3
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    • pp.116-124
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    • 2010
  • This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

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CYTOTOXIC EFFECT OF RETROGRADE FILLING MATERIALS INCLUDING GLASS IONMER CEMENT ACCORDING TO CELL LINES AND ASSAY METHODS (광중합형 glass ionomer cement를 포함한 수종 역충전재의 세포주와 검사법에 따른 독성 효과)

  • Im, Mi-Kyung;Koo, Dae-Hoi
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.403-424
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    • 1996
  • Cell culture methods have been used to assess the cytotoxicity of dental materials. Different paramaters are used to monitor cytotoxic effects. But it is difficult to compare each investigator's results with different methods. The objective of this study was to investigate cytotoxic effect of several retrograde filling materials according to cell lines and assay methods. Cytotoxicity of Bestalloy (Dogmyung, Korea), Prisma APH(Densply International Inc., U.S.A.), Clearfil FII (Kuraray Co., Japan), Fuji II (GC Co., Japan), Fuji II LC (GC Co., Japan) and IRM (Densply Co., U.S.A.) on L929, 3T3 and KB permanent cell lines was measured. Radiochromium, Lactate dehydrogenase (LDH) release method and colorimetric assays, namely neutral red (NR) and MTT were used. Each material was mixed according to the manufacturer's instruction. They were tested as solid and extracted state. Cell culture media were added to each mixed or solid materials then the solution was collected and used as extract solutions. Solid Fuji II showed mild cytotoxicity on three cell lines using radiochromium release method. There was no difference in cytotoxicity of extract solution group using radiochromium release method. In colorimetric assay immediate Fuji II group and all the IRM groups showed severe cytotoxic effect. Difference in cyctotoxicity was due to rather kinds of cell lines than assay methods. Solid Fuji II and IRM showed mild cytotoxicity on three cell lines. But extract solutions had different cytotoxic effect according to cell lines using LDH release assay. Light-cured glass ionomer had mild to moderate degree of cytotoxicity on three cell lines. Cytotoxicity was affected by specimen prepaton. Susceptibility of each cell ines were also affected by assay emthods. It was suggested that cytotoxicity study using only one cell line and/or assay method might not accurately reflect the real toxic nature of dental biomaterials.

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The Growth Inhibitory Effects of Epigallocatechin Gallate Against Human Skin Melanoma Cells and Human Oral Epitheloid Carcinoma Cells (Epigallocatechin gallate의 인체 피부흑색종세포와 인체 구강유상피암종세포에 대한 성장억제효과)

  • 한두석;박승택;백승화
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.98-103
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    • 1998
  • Epigallocatechin gallate (EGCG) was reported to exert weak cytotoxicity against normal healthy cells such as C3H10T1/2 cells, but profound inhibitory effects on the initiation or promotion stage of chemical carcinogenesis in mammary gland, blood and mouse skin. This study was carried out to develop antitumor agents with weak side effects and strong antitumor activity. Human skin melanoma cells (HBT 69) and human oral epitheloid carcinoma cells (OCL 17) were cultured in RPMI-1640 media containing 10% fetal bovine serum, antibiotic, and fungizone. After incubation for 24 hrs, the cells were treated with various amounts of (EGCG) for 48 hrs. The growth inhibitory effects of EGCG in human oral epitheloid carcinoma cells were evaluated by the 3- (4,5-djmethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR), and sulforhodamine B protein (SRB) assays of colorimetric methods. The light microscopic study was also carried out to observe morphological changes of the treated cells. These results obtained were as follows; 1. Significantly inhibitory effects of EGCG against cultured human oral epithelioid carcinoma cells. 2. Significantly inhibitory effects against cultured human skin melanoma cells treated with 50 $\mu$M EGCG, but decreased inhibitory effects in 100 $\mu$M EGCG. 3. Degenerative changes against cultured human oral epitheloid carcinoma cells. 4. Degenerative changes against human skin melanoma cells treated with 50 UM EGCG, but recovered degenerative changes in 100 $\mu$M EGCG.

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Colorimetric Evaluation of the Time-Killing Assay for Citropin 1.1, Lipopeptide Palm-KK-$NH_2$, and Temporin A

  • Baranska-Rybak, Wioletta;Dawgul, Malgorzata;Bielinska, Sylwia;Kraska, Bartlomiej;Piechowicz, Lidia;Kamysz, Wojciech
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.536-539
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    • 2011
  • Nowadays, there are a number of colorimetric techniques available for the determination of a time killing assay in a manner much easier and faster than those previously more commonly used, which were much more time-consuming and laborious colony counting procedures. Here, an attempt has been made to test the antimicrobial peptides of Citropin 1.1, Palm-KK-$NH_2$, and Temporin A on a reference strain of Staphylococcus aureus using resazurin as the cell viability reagent. Staphylococcus aureus was exposed to the test compounds over varying periods of time and the metabolic activity measured, with a profile of antimicrobial activity then established. The results are in agreement with data from previous literature, thus confirming the relevance of the application of resazurin for the testing of antimicrobial agents.

Enzyme-linked Immunosorbent Assay Strip Sensor for Rapid Detection of Staphylococcus aureus (Staphylococcus aureus 신속 검출을 위한 효소면역측정 스트립 센서)

  • Park, So Jung;Kim, Young-Kee
    • Applied Chemistry for Engineering
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    • v.22 no.5
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    • pp.522-525
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    • 2011
  • In this study, an established enzyme-linked immunosorbent assay and immuno-chromatography technique are combined to fabricate an immuno-strip sensor for the detection of S. aureus. The immuno-strip is manufactured by using four different functional membranes. The capture antibody is immobilized on the nitrocellulose membrane due to the high affinity and the capillary action through porous membranes induces a flow of sample. A colorimetric signal is appeared according to the enzyme reaction and is analyzed by the digital camera (qualitative analysis) and home-made image analysis software (quantitative analysis). Under the optimal conditions, samples with S. aureus in the range of $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ can be detected by the colorimetric method within 30 min.

Phenolic Compounds from Duchesnea chryszntha and their Cytotoxic Activities in Human Cancer Cell

  • Lee, Ihn-Rhan;Yang, Mi-Young
    • Archives of Pharmacal Research
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    • v.17 no.6
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    • pp.476-479
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    • 1994
  • Five pohenolic compounds were isolated from 80% aq. acetone extract of Duchesnea chyrysantha. Their crytotoxicities were screened by the colorimetric tetrazolium assay (MIT assay). Gallic acid, methyl caffeate, protocatechuic acid and pedunculagin mildly inhibited the survival of $PC_{14}{\;}and{\;}MKN_{45}$ human cancer cell. Brevifolin carboxylic acid showed a strong cytotoxic activity.

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The Inhibitory Effects of Taraxaci Herba against Cadmium induced Cytotoxicity (포공령의 카드뮴에 대한 세포독성 억제효과)

  • Han, Du-Seok;Lee, Ki-Nam;Lee, Jong-Sub;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.307-311
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    • 1998
  • This study was carried out to evaluate antitoxic effects Taraxaci Herba extract against Cadium by calorimetric methods. The antitoxic activity of Taraxaci Herba ex tract in NIH 3T3 fibroblasts was evaluated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-phenyl-2H-tetrazoliumbromide), NR (Neutral red) and SRB (Sulforhodamine B protein) assay. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of $10^{-2}mg/ml$ of Taraxaci Herba extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were antitoxic and tend to regenerate. These results suggest that Taraxaci Herba extract retains a potential antitoxic activity.

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A STUDY ON THE CYTOTOXIC EFFECTS OF MITOMYCIN C AND 5-FLUOROURACIL IN CULTURED RAT FIBROBLASTS

  • C. S. M;Park, Hong-Seog;Chung, Yeun-Tai
    • Toxicological Research
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    • v.7 no.1
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    • pp.13-20
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    • 1991
  • To investigate the cytotoxicity and genotoxicity of the DNA alkylating agnet, mitomycin C and the antimetabolite, 5-Fluorouracil (5-FU) in cultured rat fibroblasts, the colorimetric assay of netural red (NR) for cytotoxicity and for genotoxicity, sister chromatid exchange (SCE) assay and the measurement of the rate of DNA synthesis were performed in cells cultured in media containing various concentrations of mitomycin C and 5-FU. The uptake ability of neutral red decreased does-dependently. NR90 and NR50 values of mitomycin C were 1.49 nM and 6.87mM and 5-FU were 38.4mM AND 284.4Mm respectively.

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Salicylimine-Based Colorimetric and Fluorescent Chemosensor for Selective Detection of Cyanide in Aqueous Buffer

  • Noh, Jin Young;Hwang, In Hong;Kim, Hyun;Song, Eun Joo;Kim, Kyung Beom;Kim, Cheal
    • Bulletin of the Korean Chemical Society
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    • v.34 no.7
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    • pp.1985-1989
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    • 2013
  • A simple colorimetric and fluorescent anion sensor 1 based on salicylimine showed a high selectivity and sensitivity for detection of cyanide in aqueous solution. The receptor 1 showed high selectivity toward $CN^-$ ions in a 1:1 stoichiometric manner, which induces a fast color change from colorless to orange and a dramatic enhancement in fluorescence intensity selectively for cyanide anions over other anions. Such selectivity resulted from the nucleophilic addition of $CN^-$ to the carbon atom of an electron-deficient imine group. The sensitivity of the fluorescence-based assay (0.06 ${\mu}M$) is below the 1.9 ${\mu}M$ suggested by the World Health Organization (WHO) as the maximum allowable cyanide concentration in drinking water, capable of being a practical system for the monitoring of $CN^-$ concentrations in aqueous samples.