• Title/Summary/Keyword: MTT method.

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Antineoplastic Effect of Extracts from Traditional Medicinal Plants (전통 약용식물의 항암효과에 대한 연구)

  • Park, Jae-Gahb;Hyun, Jin-Won;Lim, Kyoung-Hwa;Shin, Jin-E;Won, Yong-Jin;Yi, Young-Deuk;Shin, Kuk-Hyun;Chang, Il-Moo;Woo, Won-Sick
    • Korean Journal of Pharmacognosy
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    • v.24 no.3
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    • pp.223-230
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    • 1993
  • Antineoplastic activity against human gastric, colon and hepatocellular carcinoma cell lines were measured in 100 extracts from 80 medicinal plants using MTT (3-[4, 5-dimethyl thiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) method. Seventeen extracts from fourteen plants, all of which have previously been reported to have antineoplastic effect, had $IC_{50}$(50% inhibitory concentration) values of less than $230{\;}{\mu}g/ml$ in at least one of the three cell lines. Extracts from remaining sixty-six medicinal plants failed to show significant cytotoxic effect at the concentration of less than $230{\;}{\mu}g/ml$.

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The Effects on Antimicrobial and Cytotoxicity of Solanum Iyratum Fractions (배풍등 분획물의 항균 및 암세포 증식 억제효과)

  • Shin Mi-Ok
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.948-954
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    • 2005
  • In this study, we investigated antimicrobial and cytotoxicity effects to each fraction extracted from Solanum lyratum (SL), which were extracted methanol (SLM) and then the extract was fractionated into five different types : hexane (SLMH), ethyl ether (SLMEE), ethylacetate (SLMEA), butanol (SLMB) and aqueous (SLMA). The antimicrobial activity was analyzed by the paper disc method. Among the various solvent fractions, SLMEA showed the strongest antimicrobial activies. The cytotoxicity of SL fractions on HeLa, MCF-7, HT-29 and HepG2 cells was evaluated by MTT assay. Among various partition layers, SLMEE showed the strongest cytotoxic effects to all cancer cell lines. We also observed that quinone reductase (QR) was induced by all fraction layers of SL to HepG2 cells. Since the QR-induced effects of SLMEE on HepG2 cells at $160{\mu}g/ml$ concentration showed 2.1 when compared with a control value of 1.0, inducer of QR for cancer protection may be contained in this fraction.

Development of assay method for the activities of new compounds, and the effect of several fungicides against spore germination, adhesion, and myceial growth of Colletotrichum sp. causing red pepper anthracnose (고추 탄저병균의 포자 발아와 부착, 균사 생장에 미치는 화합물의 활성 검정법 확립 및 살균제의 효과)

  • Kim, Jae-Jeung;Kim, Joon-Tae;Park, Sung-Woo;Park, Eun-Suk;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.7 no.3
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    • pp.159-168
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    • 2003
  • With microtiter plate, the assay method was developed for detecting the fungicidal activity of new compounds against spore germination, spore adhesion and mycelial growth of Colletotrichum sp. JC24 cal1Sing red pepper anthracnose. Also, the effects of some commercialized fungicides on fungal development like above mentioned were investigated by measuring the optical density of mycelia grown into wells of microtiter plate. For the standardization of assay method, some factors, such as the treatment of MTT and/or propanol, inodulum density and incubation period, affecting on mycelial optical density were investigated. For obtaining precise and consistent mycelial optical density, it was necessary the treatment of MTT for 12 hrs and propanol for 1 hr. inoculum density adjusted to $1\times10^5$ spores/mL and incubation period for 36 hrs at $25^{\circ}C$. For fungicidal activities, 6 protective fungicides, 6 ones inhibiting sterol biosynthesis, and one inhibiting respiration were used in this study. While mancozeb, chlorothalonil and dithianon among 6 protective fungicides inhibited strongly spore germination, adhesion, and mycelial growth at $6.25{\mu}g/mL$, propineb, iminoctadine and fluazinam inhibited intermediately spore germination and mycelial growth at $100{\mu}g/mL$. Washing above 3 fungicides with new PD broth, their activity against spore adhesion decreased. With hexaconazole, tebuconazole and myclobutanil, the tendency of the activity against fungal differentiation of the early infection stage was similar to the latter group of protective fungicides, showing the decrease of the inhibitory activity against spore adhesion by washing 2 hrs after incubation. However, kresoxim-methyl inhibited spore adhesion distinctly, depending on the applied concentrations. Based on these results, it might be able to assess the fungicidal activity of many compounds against spore germination, adhesion and mycelial growth by the use of microtiter plate in vitro. Using the assay developed in this report, it was possible to investigate the inhibitory activity of some commercialized fungicides, too.

Celecoxib, a COX-2 Selective Inhibitor, Induces Cell Cycle Arrest at the G2/M Phase in HeLa Cervical Cancer Cells

  • Setiawati, Agustina
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1655-1659
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    • 2016
  • Celecoxib, a selective inhibitor of COX-2, showed cytotoxic effects in many cancer cell lines including cervical cancer cells. This study investigated the effect of celecoxib on cell cycle arrest in HeLa cervical cancer cells through p53 expression. In vitro anticancer activity was determined with the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) method. A double staining method was applied to investigate the mechanism of cell death, cell cycling was analyzed by flow cytometryand immunocytochemistry was employed to stain p53 expression in cells. Celecoxib showed strong cytotoxic effects and induced apoptosis with an $IC_{50}$ value of $40{\mu}M$. It induced cell cycle arrest at G2/M phase by increasing level of p53 expression on HeLa cells.

A cost-effective and simple culture method for primary hepatocytes

  • Adaya, Sezin;Hasircib, Nesrin;Gurhana, Ismet Deliloglu
    • Animal cells and systems
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    • v.15 no.1
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    • pp.19-27
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    • 2011
  • Hepatocytes, the major epithelial cells of the liver, maintain their morphology in culture dishes coated with extracellular matrix (ECM) components such as collagen and fibronectin or biodegradable polymers (e.g. chitosan, gelatin). In these coated dishes, survival of cells and maintaining of liver-specific functions may increase. The aim of this study was to determine a suitable, cost-effective and simple system for hepatocyte isolation and culture which may be useful for various applications such as in vitro toxicology studies, hepatocyte transplantation and bioartificial liver (BAL) systems. In order to obtain primary cultures, hepatocytes were isolated from liver by an enzymatic method and cultured on plates coated with collagen, chitosan or gelatin. Collagen, gelatin-sandwich and gelatin-cell mixture methods were also evaluated. Morphology and attachment of the cells were observed by inverted microscope and scanning electron microscope (SEM). An MTT assay was used to determine cell viability and mitochondrial activity.

Effects of Cytotoxic and Antioxidant of Methanol Extracts from Medicinal Plants (어성초 돌콩 추출물의 항암활성 및 항산화 활성 효과)

  • Lee, Jeong-Ho;Kim, Yun-Gyoung;Choi, Choi-Mun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.18 no.3
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    • pp.37-43
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    • 2005
  • This study was performed to determine the cytotoxic effect of methanol extract from Houttuynia Cordata and Glycine soja. The cell viability was determined by MTT method. Their cytotoxic activities against three cancer cell lines such as A549, MDA-MB-231 and SNU-C4 cell line were tested. Among them, The methanol extract of Houttuynia Cordata showed the strongest cytotoxic effect against SNU-C4 cells. These results suggest that the methanol extract of Houttuynia Cordata possessed a potential antitumorous agent. The free radical scavenging activity using DPPH method was the strongest of Houttuynia Cordata methanol extract and ethyl acetate fraction.

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Upregulation of Mir-34a in AGS Gastric Cancer Cells by a PLGA-PEG-PLGA Chrysin Nano Formulation

  • Mohammadian, Farideh;Abhari, Alireza;Dariushnejad, Hassan;Zarghami, Faraz;Nikanfar, Alireza;Pilehvar-Soltanahmadi, Yones;Zarghami, Nosratollah
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8259-8263
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    • 2016
  • Background: Nano-therapy has the potential to revolutionize cancer therapy. Chrysin, a natural flavonoid, was recently recognized as having important biological roles in chemical defenses and nitrogen fixation, with anti-inflammatory and anti-oxidant effects but the poor water solubility of flavonoids limitstheir bioavailability and biomedical applications. Objective: Chrysin loaded PLGA-PEG-PLGA was assessed for improvement of solubility, drug tolerance and adverse effects and accumulation in a gastric cancer cell line (AGS). Materials and Methods: Chrysin loaded PLGA-PEG copolymers were prepared using the double emulsion method (W/O/W). The morphology and size distributions of the prepared PLGA-PEG nanospheres were investigated by 1H NMR, FT-IR and SEM. The in vitro cytotoxicity of pure and nano-chrysin was tested by MTT assay and miR-34a was measured by real-time PCR. Results: 1H NMR, FT-IR and SEM confirmed the PLGA-PEG structure and chrysin loaded on nanoparticles. The MTT results for different concentrations of chrysin at different times for the treatment of AGS cell line showed IC50 values of 68.2, 56.2 and $42.3{\mu}M$ and 58.2, 44.2, $36.8{\mu}M$ after 24, 48, and 72 hours of treatment, respectively for chrysin itslef and chrysin-loaded nanoparticles. The results of real time PCR showed that expression of miR-34a was upregulated to a greater extent via nano chrysin rather than free chrysin. Conclusions: Our study demonstrates chrysin loaded PLGA-PEG promises a natural and efficient system for anticancer drug delivery to fight gastric cancer.

AN EXPERIMENTAL STUDY OF THE DEGREE OF CONVERSION AND CYTOTOXICITY OF DUAL CURE RESIN CEMENTS (수종 이원중합 레진 씨멘트의 중합률 및 세포 독성에 관한 실험적 연구)

  • Roh, Byoung-Duck;Park, Seong-Ho;Lee, Chung-Suck
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.33-54
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    • 1995
  • The degree of conversion of composite resin was known to have influence on the mechanical properties of composite materials such as hardness, strength, wear resisitance, dimensional and color stability. Also unreacted monomer was reported to be harmful to the pulp. So the degree of conversion was a very important factor in the success of composite resin restorations. In recent, the dual cure resin cement was developed with the advocations that it could increase the curing rates in the sites where the curing ligt could not reach. Moreover many manufactors added some adhesive components in the resin cement. This study was undertaken to observe the effects of curing depth and light curing times on the degree of conversion of dual cure resin cements. CR INLAY CEMENT, DUAL CEMENT and OPTEC BOND, by the Fourier transform Infrared analysis, changing the curing depth 1mm, 2mm and 3mm, and varying the light curing time 20 seconds, 40 seconds and 80 seconds at each depth. The cytotoxicity of dual cure resin cements was tested by the in vitro MTT method using L929 cell. The results was evaluated and compared statistically. The results were obtained as follows : 1. The dual cure resin cements reavealed various degree of conversion, CR INLAY CEMENT and DUAL CEMENT had a tendency to be more reactive to the light cure and OPTEC BOND was a more chemical one. 2. CR INLAY CEMENT and DUAL CEMENT showed the lowest degree of conversion in 2 mm depth, and in 3mm depth the degree of conversion increased, which were due to the chemical cure of dual cures, but OPTEC BOND showed decreasing degree of conversion with increasing curing dept h and all experimental groups showed lower degree of conversion than CHEMICAL group which cured in dark room with no light, so the weak light-curing of dual cure resin cement prevented the chemical cure. (P<0.05) 3. CR INLAY CEMENT and DUAL CEMENT showed increasing degree of conversion in 1 mm and 3 mm, according to the increasing cure times, but in 2 mm depth the degree of conversion decreased with increasing light-curing times and OPTEC BOND showed contrary tendency, but there was no ststistical importance in the differences among the experimental group.(P>0.05) 4. The optical density by MTT assay of extractions of CR INLAY CEMENT, DUAL CEMENT and OPTEC BOND revealed no statitically important differences comparing with optical density of negative control.(P>0.05) 5. CR INLAY CEMENT showed a tendency of increaing cytotoxicity with days and DUAL CEMENT and OPTEC BOND showed higher cytotoxicity in 2 days than in 4 days, but there was no statistical importance in the differences.(P>0.05).

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Determination of Bioactive Compounds and Anti-cancer Effect from Extracts of Korean Cabbage and Cabbage (배추와 양배추 추출물의 생리활성 물질 및 암세포 증식 억제효과 분석)

  • Hwang, Eun-Sun;Hong, Eun-Young;Kim, Gun-Hee
    • The Korean Journal of Food And Nutrition
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    • v.25 no.2
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    • pp.259-265
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    • 2012
  • In this study, we determined total polyphenol content(TPC) and total flavonoid content(TFC) of extracts from Korean cabbage and cabbage using a spectrophotometric method as well as glucosinolates concentration by HPLC. TPCs of Korean cabbage and cabbage extracts were 308.48 ${\mu}g$ GAE/g dry weight and 344.75 ${\mu}g$ GAE/g dry weight, respectively. TFCs of Korean cabbage and cabbage extracts were 5.33 ${\mu}g$ QE/g dry weight and 5.95 ${\mu}g$ QE/g dry weight, respectively. We found six different glucosinolates, namely progoitrin, glucoalyssin, gluconapin, glucobrassicanapin, glucobrassicin and 4-methoxyglucobrassicin in the Korean cabbage extract. In the cabbage extract, there was four glucosinolates, namely glucoraphanin, sinigrin, glucobrassicin and 4-methoxyglucobrassicin. We determined the cytotoxic effect of Korean cabbage and cabbage extracts in AGS human stomach cancer cells, HepG2 human hepatic cancer cells and LNCaP human prostate cancer cells by MTT assay. Dose-dependent relationships were found between the extract concentrations and cancer cell growth inhibition. The overall results support that both Korean cabbage and cabbage, the major vegetables in Korea, contain bioactive compounds such as polypheol, flavonoids as well as glucosinolates and they may play a positive role in cancer prevention.

Adhesion of Human Intervertebral Disk Cells on Aiginate/PLGA Microspheres (Alginate/PLGA 미립구에 대한 인간디스크 세포 부착 효과)

  • Lee, Jun-Hee;Jang, Ji-Wook;So, Jeong-Won;Choi, Jin-Hee;Park, Jong-Hak;Ahn, Shik-Il;Son, Young-Suk;Min, Byoung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.7-12
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    • 2009
  • PLGA microspheres have been known as an injectable system for tissue engineering. The purpose of this study was to investigate the condition of emulsion formation and cell adhesion on the microsphere surface. BSA-loaded PLGA microsphere was fabricated by oil-in-water (O/W) and water-in-oil-in-water (W/O/W) solvent evaporation method. Sodium alginate was dissolved in water phase to control initial burst release and to improve lag time by PLGA bulk degradation. In addition, the morphology of cells attached on the micro spheres was studied using a scanning electron microscopy (SEM). Cellular proliferation behavior of human disc cells cultivated on PLGA micro spheres was analyzed using a MTT assay. MTT assay revealed that the cells can attach and proliferate on PLGA microspheres. According to these results, we concluded that BSA -loaded alginate/PLGA microspheres can be used as an injectable system for tissue engineering application.