• Title/Summary/Keyword: MTT assay

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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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Therapeutic effects of dihydroartemisinin and transferrin against glioblastoma

  • Kim, Suk Hee;Kang, Seong Hee;Kang, Bo Sun
    • Nutrition Research and Practice
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    • v.10 no.4
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    • pp.393-397
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    • 2016
  • BACKGROUND/OBJECFTIVES: Artemisinin, a natural product isolated from Gaeddongssuk (artemisia annua L.) and its main active derivative, dihydroartemisinin (DHA), have long been used as antimalarial drugs. Recent studies reported that artemisinin is efficacious for curing diseases, including cancers, and for improving the immune system. Many researchers have shown the therapeutic effects of artemisinin on tumors such as breast cancer, liver cancer and kidney cancer, but there is still insufficient data regarding glioblastoma (GBM). Glioblastoma accounts for 12-15% of brain cancer, and the median survival is less than a year, despite medical treatments such as surgery, radiation therapy, and chemotherapy. In this study, we investigated the anti-cancer effects of DHA and transferrin against glioblastoma (glioblastoma multiforme, GBM). MATERIALS/METHODS: This study was performed through in vitro experiments using C6 cells. The toxicity dependence of DHA and transferrin (TF) on time and concentration was analyzed by MTT assay and cell cycle assay. Observations of cellular morphology were recorded with an optical microscope and color digital camera. The anti-cancer mechanism of DHA and TF against GBM were studied by flow cytometry with Annexin V and caspase 3/7. RESULTS: MTT assay revealed that TF enhanced the cytotoxicity of DHA against C6 cells. An Annexin V immune-precipitation assay showed that the percentages of apoptosis of cells treated with TF, DHA alone, DHA in combination with TF, and the control group were $7.15{\pm}4.15%$, $34.3{\pm}5.15%$, $66.42{\pm}5.98%$, and $1.2{\pm}0.15%$, respectively. The results of the Annexin V assay were consistent with those of the MTT assay. DHA induced apoptosis in C6 cells through DNA damage, and TF enhanced the effects of DHA. CONCLUSION: The results of this study demonstrated that DHA, the derivative of the active ingredient in Gaeddongssuk, is effective against GBM, apparently via inhibition of cancer cell proliferation by a pharmacological effect. The role of transferrin as an allosteric activator in the GBM therapeutic efficacy of DHA was also confirmed.

Properties of Fucoidan as Raw Materials of Water-holding Cream and Cosmetics (후코이단의 보습제 및 화장품재료로서 특성)

  • Cha, Sung Hwan;Lee, Jung Shik;Kim, Young Suk;Kim, Donguk;Moon, Jae Cheon;Park, Kwonpil
    • Korean Chemical Engineering Research
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    • v.48 no.1
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    • pp.27-32
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    • 2010
  • Properties of fucoidan used for functional cosmetics agents were studied. Fucoidan was extracted from Undaria pinnatifida sporophylls. To test possibility of fucoidan as a cosmetics material, water-holding property measurement, DPPH free radical scavenging assay and MTT assay were done. Water-holding property of fucoidan was higher than that of hyaruronic acid, which is known as the one of the best water-holding material. The water-holding strength of fucoidan slightly increase as molecular weight of fucoidan decrease. Fucoidan showed high stability from MTT assay and good anti-oxidation property from DPPH assay. To evaluate the effect of water-holding property and anti-alergy property of fucoidan on the atopic dermatitis(AD), 46 AD patients were treated with fucoidan cream. After 6 weeks treatment, Investigation Global Assessment(IGA) scores decreased from 3.04 to 2.15, that is fucoidan cream had a 39.8% benefit effect on atopic dermatitis.

Evaluation of the Cytotoxicity of Calcium Phosphate Root Canal Sealers (인산 칼슘 근관 봉함재의 세포독성에 관한 연구)

  • Lee, Jee-Hyun;Baek, Seung-Ho;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.28 no.4
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    • pp.295-302
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    • 2003
  • 본 연구에서는 인산 칼슘 근관 봉함재 [Apatite Root Sealer (ARS) Type I, II, III]의 세포독성을 혼합후의 시간 경과에 따라서 다른 4계열의 근관 봉함재 (Pulp Canal Sealer EWT, AH Plus, Sealapex, Ketac Endo)와 비교하여 평가하였다. 근관 봉함재를 혼합한 후 1시간, 8시간, 24시간, 48시간, 1주, 2주,4주의 기간동안 배양액을 이용하여 추출액을 얻었다. L929 쥐섬유아세포를 24시간동안 각 시간군에서 얻은 추출액과 함께 배양한 후 dimethylthiazol diphenyltetrazolium (MTT) assay와 neutral red (NR) assay를 이용하여 세포독성(%)을 평가하였다. 실험 결과 ARS Type I, II, III는 전 시간군에 걸쳐 낮은 세포독성을 보였고 (23.65-0.55%) 특히 경화 초기에 다른 계열의 근관 봉함재보다 유의성 있게 낮은 세포독성을 나타내었다. ARS Type I, II, III간의 세포독성은 각 시간군에서 유의성 있는 차이를 보이지 않았다. AH Plus와 Ketac Endo는 초기에 높은 세포독성을 보였으나 AH Plus는 8시간 이후에, Ketac Endo는 24시간 이후에 독성이 급격히 감소하여 ARS와 유의성 있는 차이를 보이지 않았다. Pulp Canal Sealer EWT와 Sealapex는 4주까지 지속적인 세포독성을 나타내었다. 그리고 MTT assay와 NR assay를 이용하여 얻은 각 근관봉함재의 세포독성은 시간 경과에 따라서 비슷한 변화 양상을 나타내었다. 인산 칼슘 근관 봉함재는 생체적합성이 우수한 재료로써 앞으로 지속적인 개발과 평가가 필요할 것으로 사료된다.

Effects of Taeumin Chungsimyeunjatang on the Cerebral neurons injured by Hydrogen Peroxide (태음인(太陰人) 청심연자탕(淸心蓮子湯)이 Hydrogen Peroxide에 손상(損傷)된 백서(白鼠)의 대뇌신경세포(大腦神經細胞)에 미치는 영향(影響))

  • Ok, Yun-young;Ryu, Do-gon;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.251-266
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    • 1999
  • 1. Purpose : The purpose of this study was to determine the effects of Chungsimyeunjatang on the cerebral neurons injured by hydrogen peroxide($H_2O_2$). 2. Methods : I observed cell viability in mouse cerebral neurons exposed to hydrogen peroxide by NR assay and MTT assay and determined lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. After administration of Chungsimyeunjatang water extracts, I observed significant changes of cell viability, lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide. 3. Results : Hydrogen peroxide showed neurotoxicity. Cell viability in mouse cerebral neurons exposed to hydrogen peroxide decreased in NR assay and MTT assay. Lipid peroxidation and amounts of LDH release in mouse cerebral neurons exposed to hydrogen peroxide increased. Chungsimyeunjatang was very effective in blocking hydrogen peroxide-induced neurotoxicity.

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Effects of Yuldahansotang water extract on Cultured Spinal Sensory Neurons Damaged by Xanthine Oxidase/Hypoxanthine (열다한소탕(熱多寒少湯) 전탕액(煎湯液)이 XO/HX에 의해 손상(損傷)된 배양척수감각신경세포(培養脊髓感覺神經細胞) 미치는 효과(效果))

  • Hong, Jeong-a;Kim, Kyung-yo;Yu, Do-gon;Park, Hye-sun;Kim, Hyung-soon
    • Journal of Sasang Constitutional Medicine
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    • v.13 no.1
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    • pp.88-96
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    • 2001
  • To evaluate the effect of Yuldahansotang(YHT) water extract on cultuted mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MIT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried our after the cultured mouse spinal sensory neuron were preincubated with various concentrations of YHT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cells on NR assay and MTT assay. 2. MTT50 value and NR50 value pf XO/HX were 20 mU/ml XO/0.2 mM HX and 40 mU/ml XO/0.2 mM HX. 3. YHT water extract have efficacy of increasing neurofilament. 4. YHT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that YHT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

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Effects of Several Salt Marsh Plants on Mouse Spleen and Thymus Cell Proliferation Using MTT Assay

  • Seo, Young-Wan;Lee, Hee-Jung;Kim, You-Ah;Youn, Hyun-Joo;Lee, Burm-Jong
    • Ocean Science Journal
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    • v.40 no.4
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    • pp.209-212
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    • 2005
  • In the present study, we have tested the effects of 21 salt marsh plants on cell proliferation of mouse immune cells (spleen and thymus) using MTT assay in culture. The methanolic extracts of six salt marsh plants (Rosa rugosa, Ixeris tamagawaensis, Artemisia capillaris, Tetragonia tetragonoides, Erigeron annus, and Glehnia littoralis) showed very powerful suppressive effects of mouse immune cell death and significant activities of cell proliferation in vitro. Especially, the methanolic extract of Rosa rugosa was found to have fifteen times compared to the control treatment, demonstrating that Rosa rugosa may have a potent stimulation effect on immune cell proliferation. These results suggest that several salt marsh plants including Rosa rugosa could be useful for further study as an immunomodulating agent.

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.

Development of Anticancer Agents from Korean Medicinal Plants. Part 7. Cytotoxic Activity of the Chloroform soluble Fraction of Perrila frutescens Against Human Oral Epitheloid Carcinoma Cells (한국산 생약으로 부터 항암물질의 개발 (제7보), 소엽의 Chloroform 가용성 분획이 인체 구강유상피암종세포에 미치는 세포독성작용)

  • Han, Du-Seok;Kim, Young-Il;Choi, Kyw-Eun;Kwag, Jung-Suk;Baek, Seung-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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    • pp.37-42
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    • 1998
  • In the present study, we have evaluated cytotoxic effects of the chloroform soluble fraction of the methanolic extract of Perilla frutescens in human oral epitheloid carcinoma cells. The light microscopic study showed morphological changes of the treated cells. Cell membrane damaging activity was measured by the lactate dehydrogenase (LDH) assay and disruptions in cell organelles were determined by 3-(4,5-dime-thylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), neutral red (NR) and sulforhodamine protein B (SRB) of colorimetric assay. These results suggest that Perilla frutescens retains a potential antitumor activity.

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Effects of Yuldahansotang water extract on Cultured Primary Hippocampal Cell Culture Damaged by Hydrogen Peroxide (열다한소탕(熱多寒少湯) 전탕액(煎湯液)이 Hydrogen Peroxide에 의해 손상(損傷)된 배양(培養) 해마신경세포(海馬神經細胞)에 미치는 영향)

  • Park, Hye-Sun;Kim, Kyung-Yo;Go, Gi-Deok;Kim, Il-Hwan;Lee, Jae-Heung
    • Journal of Sasang Constitutional Medicine
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    • v.14 no.1
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    • pp.79-89
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    • 2002
  • To evaluate the effect of Yuldahansotang(YHT) water extract on cultured hippocampal cell was inhibited by hydrogen peroxide, MTT assay, NR assay, Neurofilament enzymeimmuno assay and DNA synthesis assay were carried out after the cultured hippocampal cells were preincubated with various concentrations of YHT water extract for 3 hours prior to exposure of hydrogen peroxide. The results obtained were as follows: 1. Hydrogen Peroxide decreased the survival rate of the cultured hippocampal cells on NR assay and MIT assay. 2. YHT water extract have efficacy of increasing a amount of neurofilament decreased by hydrogen peroxide in cultured hippocampal cells. 3. YHT water extract have efficacy of increasing DNA synthesis decreased by hydrogen peroxide in cultured hippocampal cells. From above the results, It is concluded that YHT has marked efficacy in preventing for the damages by hydrogen peroxide.

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