• Title/Summary/Keyword: MTT method.

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The biomedical effect of 655nm Laser Diode irradiation (655nm 레이저 다이오드 조사에 따른 생물학적 특성 평가)

  • Cheon, Min-Woo;Kim, Seong-Hwan;Park, Yong-Pil;Lee, Ho-Shik;Kim, Tae-Gon;Park, No-Bong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.11a
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    • pp.397-398
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    • 2008
  • This paper performed the basic study for fabricating the low level laser therapy apparatus, and one of the goals of this paper was to make this apparatus used handily. The apparatus has been fabricated using the 655nm laser diode and microprocessor unit. The apparatus used a 655 nm laser diode for laser medical therapy and was designed for a pulse width modulation type to increase stimulation effects. And then, each experiment was performed to irradiation group and non-irradiation group for cells. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590nm transmittance of ELISA reader. As a result, the cell increase of cells was verified in irradiation group as compared to non-irradiation group.

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Cytotoxicity and Antimicrobial Effects of Extracts from Salvia miltiorrhiza (단삼 추출물의 세포독성과 항균효과)

  • Kwag, Jung-Sook;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.34 no.4 s.135
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    • pp.293-296
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    • 2003
  • This study was carried out to evaluate cytotoxic effects of Salvia miltiorrhiza extracts on NIH 3T3 fibroblasts and KB cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The comparison of $IC_{50}$ of values of Salvia miltiorrhiza extracts in KB cell lines showed that their susceptibility to these extracts decreased in the following order: hexane extract > chloroform extract > methanol extract> dichloromethane extract > ethyl acetate extract>ethanol extract by the MTT method. The dried roots of Salvia miltiorrhiza was extracted several solvents, and then antimicrobial activity was investigated. The minimal inhibitory concentrations (MIC's) of the extract against microorganisms were also examined. Amtimicrobial activity of ketoconazol as reference was compared to those of extracts of hexane, chloroform, dichloromethane, ethyl acetate, ethanol and methanol. The antimicrobial activity of all extracts from the sample had growth inhibition activity against gram-negative bacteria, gram-positive bacteria and fungi. These results suggest that the hexane and chloroform soluble extracts of Salvia miltiorrhiza may be a valuable choice for the studies on the tumor cell lines and growth inhibition activity.

Cytotoxic Saponins from the Root of Dipsacus asper Wall

  • Hung Tran Manh;Jin WenYi;Thuong Phuong Thien;Song Kyung Sik;Seong Yeon Hee;Bae KiHwan
    • Archives of Pharmacal Research
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    • v.28 no.9
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    • pp.1053-1056
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    • 2005
  • Cytotoxic activitiy of seven hederagenin saponins isolated from the root of Dipsacus asper were investigated in vitro against L1210, HL-60 and SK-OV-3 tumor cell lines by the MTT method. $3-O-\alpha-L-rhamnopyranosyl-(1{\rightarrow}2)-\alpha-L -arabinopyranosyl$ hederagenin (2),\;$3-O-\beta-D­xylopyranosyl-( 1{\rightarrow}3)-\alpha-L-Rhamnopyranosyl-(1{\rightarrow}2)-\alpha-L -arabinopyranosyl$ hederagenin (6) and $3-O-\beta-D-glucopyranosyl-(1{\rightarrow}3)-\alpha-L-rhamnopyranosyl-( 1{\rightarrow}2)-\alpha-L-arabinopyranosyl$ hederagenin (7) exhibited the potent cytotoxicity against the three tumor cell lines with $IC_{50}$ values ranging from 4.7 to 8.7 ${\mu}g/mL$, with the exception of compound 7, which exhibited weak cytotoxic activity against SK-OV-3 $(IC_{50}\;22.5\;{\mu}g/mL)$. Other compounds did not exhibit any cytotoxic activity $(IC_{50}>30{\mu}g/mL)$.

Anti-inflammatory effect of various solvent extract from Atractylodes japonica on Lipopolysaccharide-induced Inflammation in BV2 cells. (창출 추출물의 BV2 cell 소염작용에 관한 실험적 연구)

  • Hur, Inn-Hee;Sim, Sung-Yong;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.2 s.33
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    • pp.36-46
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    • 2007
  • Objective : In this study, the effect of Atractylodes japonica against LPS induced inflammation in mouse microglia BV2 cells was investigated. Method : Microglia BV2 Cells viability was determined using the MTT assay. We used water, ethanol extract from Atractylodes japonica and studied on the anti-inflammatory effect of lipopolysaccharide-induced inflammation using reverse transcription polymerase chain reaction (RT-PCR), western blot, and nitric oxide detection on mouse microglia BV2 cells. Result : The MTT assay revealed that it's extract has no significant cytotoxicity in the microglia BV2 cell. Various solvent extract from Atractylodes japonica inhibited nitrite production, iNOS protein and mRNA expression levels. And also it's extracts significantly reduced lipopolysaccharide-induced COX-2 activation in RT-PCR and western blot in lipopolysaccharide-induced microglia BV2 cells Conclusion : In this study, it's extracts was shown to suppress NO production by inhibiting iNOS expression and COX-2 activity. With this effects of anti-inflammation, we suggests that, it's extracts may be a useful candidate for the development of a drug on the related inflammatory diseases in brain.

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Molecular-vased sensitivity of human leukemia cell line U937 to antineoplastic activity in a traditional medicinal plants(Selaginella tamariscina) (전통 약용 식물 권백(Selaginella tamariscina)의 항암효과에 대한 혈액 암세포주 U937의 감수성 및 그 작용기구에 대한 분자생물학적 연구)

  • 이인자;이인선;박성희
    • Journal of Food Hygiene and Safety
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    • v.11 no.1
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    • pp.71-75
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    • 1996
  • In order to study the antitumoral effect of Selaginella tamariscina extracts, the cytotoxicities to human histiocytic leukemia cells (U937) and lymphocyte were measured by MTT method. The water extract of Selaginella tamariscina was partitioned into chloroform (CHCl3), ethylacetate (EtAc), n-butanol (BuOH) and water (H2O), successively. CHCl3, EtAc and BuOH fractions of Selaginella tamariscina showed the cytotoxicity to the U937 cells but they had effect on the cytotoxicity of lymphocyte under the same conditions. The tumor-specific cytotoxicity of Selaginella tamariscina fractions migh have been attributed to their genotoxic effect on actively proliferating cells. The expression of p53 tumor suppressor gene was then evaluated by northern blotting. The increased expression of p53 was induced by Selaginella tamariscina fraction V but no expression of p53 was induced by CHCl3, EtAc, and BuOH fractions of Selaginella tamariscina water extract (fraction V) should be required for the cytotoxcity on U937 and the other fractions of Selaginella tamariscina mediated the U937 disruption.

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Cytoprotective Effect of Galgeuntang Extract on Cadmium-Induced Cytotoxicity (갈근탕 추출물의 카드뮴 독성에 대한 세포 보호효과)

  • Park, So-Yun;Baek, Jong-Min;Baek, Seung-Hwa
    • YAKHAK HOEJI
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    • v.54 no.3
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    • pp.151-156
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    • 2010
  • Cytotoxicity of cadmium on NIH 3T3 fibroblasts was utilized in order to discover antitoxic compound in Galgeuntang extract in this study. Treatment groups were chosen as follows; control (medium only), $MTT_{50}$ group and five experimental groups. MTT {3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide} method was performed to evaluate the cytotoxicity of cell organelles and $IC_{50}$ was also measured. Accordingly we have examined the detoxification effects of Galgeuntang extract on cadmium-treated NIH 3T3 fibroblasts to observe morphological changes by the light microscopy. Galgeuntang extract showed cytoprotective effects on cadmium-induced cytotoxicity. Furthermore, Galgeuntang showed a dose-dependency in detoxication. The phenolic content of Galgeuntang ethanol extract was higher than that of water content. These results suggest that Galgeuntang extract may be used as a cytoprotective agent against cadmium (II)-mediated cytotoxicity.

The effect of 3mW 850nm Laser Diode on RAT BM-cell (3mW 850nm Laser Diode가 Rat BM-Cell에 미치는 효과)

  • Cheon, Min-Woo;Kim, Seong-Hwan;Park, Yong-Pil
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.514-515
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    • 2008
  • Low level laser therapy has various therapy effects. This paper performed the basic study for developing the Low Level Laser Therapy Equipment for medical treatment. The apparatus has been fabricated using the laser diode and microprocessor unit. This equipment was fabricated using a micro-controller and a laser diode, and designed to enable us to control light time, frequency and so on. In this study, the designed device was used irradiation to find out how 850 nm laser diode affected the cell proliferation of RAT bone-marrow cells. Experiment was performed to irradiation group and non-irradiation group for Rat bone marrow cells. MTT assay method was chosen to verify the cell increase of two groups and the effect of irradiation on cell proliferation was examined by measuring 590 nm transmittance of micro plate reader. As a result, the cell increase of Rat bone marrow cells was verified in irradiation group as compared to non-irradiation group. The fact that specific wavelength irradiation has an effect on cell vitality and proliferation is known through this study.

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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.

Development of Anticancer Agents from Korean Medicinal Plants. Part 13. -Studies on the Cytotoxicity and Antitumor Activity of Herba crotalariae sessiliflorae- (한국산 생약으로부터 항암물질의 개발 (제13보). -농길리 추출물의 세포독성 및 항암작용에 관한 연구-)

  • Shin, Min-Kyo;Song, Ho-Joon;Kang, Young-Sung;Ryu, Hong-Sun;Han, Du-Seok;Kang, Kil-Ung;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.30 no.2
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    • pp.130-136
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    • 1999
  • The cytotoxic and antitumor activity of Herba cratalariae sessiliflorae on cultured NIH 3T3 fibroblast and human oral epitheloid carcinoma cells were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) colorimetric method. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and human oral epitheloid carcinoma cells (KB). These results were obtained as follows; Ethyl acetate, chloroform and hexane extracts showed a significant cytotoxicity in NIH 3T3 fibroblast, but the other extracts did not show. All extracts exhibited a significant antitumor activity in human oral epitheloid carcinoma cells, but ethanol extract did not show a antitumor activity. Hexane extract showed low cytotoxic effect, but exhibited the most antitumor activity. The MTT absorbance in NIH 3T3 fibroblast was significantly decreased by treatment with chloroform, ethyl acetate and hexane extracts respectively. Human oral epitheloid carcinoma cells was significantly decreasd by treatment with all extracts with the exception of ethanol extract. The difference in MTT absorbance in two cell Types was most remarkable when treated with water and hexane extracts. Cholroform and hexane extracts showed the strongest effect in growth inhibition of human oral epitheloid carcinoma cells. These results indicated that water extract possessed no cytotoxicity and a strong antitumor activity.

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CHEMOSENSITIVITY TEST OF HUMAN OSTEOSARCOMA AND EPIDERMOID CARCINOMAS USING MTT ASSAY (MTT법을 이용한 사람 골육종과 상피암 세포주들에 대한 항암제 감수성 검사)

  • Park, Sung-Oh;Shin, Hyo-Keun;Kim, Oh-Whan
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.13 no.4
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    • pp.391-404
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    • 1991
  • Three anticncer agents which are different in time or dosage dependence as well as in phase specificity, namely mitomycin and adriamycin from natural products, and widely different cancer cell lines_Four epidermoid carcinomas originated from larynx, cervix, skin and gut were used toghether with one osteosarcoma as the target cell of single and combined administration of anticancer drugs. Semiautomated tetrazolium dye assay(MTT) appears to offer an attractive option for chemosensitivity of head and neck cancers since it is a simple, valid and inexpensive method of assessing chemosensitivity for large samples in a short time. The results obtained form this study were as follows. 1. Good correlations were obtained with the results of the MTT test and those of $^3H$ thymidine uptake assay. 2. $LD_{50}$ values of HIST and St.Ca. which showed relatively high doubling time on adriamycin were $30{\mu}g/ml$ and $15{\mu}g/ml$ while those of HeLa, Hep-2 and KHOS/NP were $2.1{\mu}g/ml$, $4.8{\mu}g/ml$, and $6.8{\mu}g/ml$ respectively. 3. The $LD_{50}$ value of 5-FU on five cancer cells were very high ranging from 15mg/ml to almost indefinite number, which means 5-FU is very resistant to epidermoid carcinomas or osteosarcoma examined in this study. 4. Mitomycin was relatively effective showing 80% cancer killing effect on HeLa, 70% on St. Ca. and 50% on Hep-2 at the high concentrations used. 5. Adriamycin was the most effective showing 90% cancer cell killing effect on KHOS/NP, 98% on HeLa, 80% both on Hep-2 and St. Ca. The least susceptible cancer cells toward adriamycin was HIST having only 55% cell killing effect at the high cincentration. 6. Combined therapy of adriamycin and 5-FU was more effective than single administration in all the cases examined. Most effective synergism was observed on St. Ca. at the low concentration, showing 21 times higher than each single administration.

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