• Title/Summary/Keyword: MTT assay

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Inhibitory Effects of Fungal Metabolites Isolated from Foodstuffs on the Growth of Human Cancer Cell Lines (식품유래 곰팡이 대사산물의 항암효과)

  • Im, Hyo-Gwon;Yu, Mi-Hee;Chung, Duck-Wha;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.262-267
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    • 2006
  • Inhibitory effects of fungal metabolites isolated from foodstuffs on growth of human cancer cell lines were evaluated. Isolated strains were divided into four classes based on color (aerial, reverse), shape, and growth speed. Fungal metabolites extracted with ethyl acetate were investigated for their growth inhibition on six kinds of human cancer cells by MTT assay. Ethyl acetate extract showed high growth inhibition against all cancer cells tested, with D4 exhibiting the strongest growth inhibition effects against Kato III, AGS, Hepa1c1c7, and MDA-MB-231 cancer cells. These results suggest ethyl acetate extract from fungal metabolites as effective natural cancer therapeutic material.

Cytotoxic Effects of Partially Purified Substances from Bacillus polyfermenticus SCD Supernatant toward a Variety of Tumor Cell tines

  • Chang, Kyung-Hoon;Park, Jun-Seok;Choi, Jae-Hoon;Kim, Cheon-Jei;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.163-166
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    • 2007
  • The cytotoxic effects of partially purified substances from Bacillus polylfermenticus SCD toward a variety tumor cell lines were studied. Cytotoxic activity was determined with regard to the A549 (human lung carcinoma), AGS (human stomach adenocarcinoma), DLD-1 (human colon adenocarcinoma), HEC-1-B (human uterus adenocarcinoma), SW-156 (human kidney carcinoma), and NIH/3T3 (murine normal fibroblast) cell lines using the MTT assay. Cytotoxic substances were partially purified through Diaion HP-20 columns and extracted with methanol or other organic solvents (n-hexane, chloroform, ethylacetate, and butanol). B. polyfermenticus SCD supernatant showed up to 60% inhibition of cell viability fer all five human cancer cell lines tested. When treated with 10 mg/mL of n-hexane, chloroform, ethylacetate, and butanol extract, HEC-1-B cells showed a 25,62,35, and 63% rate of inhibition respectively, and AGS cells showed a 72, 61, 44, and 67% rate of inhibition, respectively. At a concentration of 10 mg/mL, 100% methanol Diaion HP-20 extracts showed inhibition rates of 97.0% toward A-549 cells, 98.1% toward AGS cells, 81.6% toward DLD-1 cells, 83.5% toward HEC-1-B cells, and 92.7% toward SW-156 cells. These results indicate that partially purified fractions from B. polyfermenticus SCD have the potential to inhibit not only colon cancer cells, but also lung, stomach uterus, and kidney cancer cells. Further studies are needed to characterize the cytotoxic substances released in B. polyfermenticus SCD cultures.

Antitumor Effects of Kluyveromyces marxianus TFM-7 Isolated from Kefir

  • Lee, Hyun-Jung;Nam, Bo-Ra;Kim, Jin-Man;Kim, Ji-Yeon;Paik, Hyun-Dong;Kim, Chang-Han
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.133-137
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    • 2007
  • The Strain TFM-7, Which has an antitumor effect, was isolated from Kefir and identified based on analysis using the API 50 CHL kit and 265 rDNA sequencing. Strain TFM-7 was confirmed to belong to the genus Kluyveromyces. Analysis of the 265 rDNA nucleotide sequences found strain TFM-7 to be related to Kluyveromyces marxianus. NRRL Y-828IT. K. marxianus. TFM-7 was cultured with potato dektrose broth medium at $27^{\circ}C$ for 72 hr, and its inhibition effects on the proliferation of seven tumor cell lines and a normal cell line were assessed using the MTT assay. The antitumor effects and growth characteristics of K. marxianus TFM-7 were investigated during a culture period of 7 days. By the $3^{rd}\;day$, K. marxianus TFM-7 showed a dry cell weight 2.39 g/L, a pH of 4.39, an ethanol content of 0.89%, and an inhibition effect on the proliferation of seven tumor cell lines above 50%, except for A-549 tumor cell line. K. marxianus TFM-7 was the most effective at inhibiting the growth of Hep-2 cell line among all tumor cell lines tested. Growth inhibition of a normal cell line, NIH/3T3, was less than 35%, suggesting a decreased level of cytotoxicity toward normal cells. These results indicate that K. marxianus TFM-7 may have used as a yeast strain with antitumor activity.

Effect of Saponin with Antioxidant Activity on Matrix Metalloproteinase in Human Dermal Fibroblasts (항산화 효능을 가진 사포닌이 사람섬유아세포에서 기질 금속 단백질 분해효소에 미치는 영향)

  • Park, Hye-Jung;Kim, Moon-Moo;Lee, Dong-Hwan
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1266-1273
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    • 2011
  • Saponin is a main component of ginseng widely known as an oriental traditional medicinal ingredient. A variety of biological effects of saponin has been reported, but its action related to skin regeneration has remained unclear so far. In this study, the effect of saponin on matrix metalloproteinase as well as its antioxidant effect in cell free system was examined in human dermal fibroblasts. First of all, as a result of investigating the effect of saponin on cell viability using MTT assay, it was shown to increase cell viability below 10 ${\mu}g$/ml, but it also showed cytotoxicity above 25 ${\mu}g$/ml. The antioxidant effect of saponin was exerted by inhibition of $H_2O_2$ in addition to reducing power above 1 ${\mu}g$/ml. In particular, saponin showed a protective effect on DNA oxidation. Furthermore, it was observed that saponin activates MMP-2 and increases MMP-1 activity in gelatin and casein zymography analyses, respectively, indicating that saponin could have potential a therapeutic agent for anti-aging and skin regeneration.

Study on Cosmeceutical Activity of four Kinds of Korea Herb Medicine Materials (4가지 한방 소재(행인, 호장근, 자초, 강황)의 화장품약리활성에 관한 연구)

  • Kim, Youngh-Hun;Lee, Su-Mi;Cheon, Soon-Ju;Jang, Min-Jung;Jun, Dong-Ha;Choi, Hyang-Ja;Cho, Woo-A;Lee, Jin-Tae
    • Journal of the Korean Society of Fashion and Beauty
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    • v.5 no.4
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    • pp.130-138
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    • 2007
  • The aim of this study was to assess the cosmeceutical activies of four kinds of Korea herb medicine extracts using in cosmetics and related industries. The cosmeceutical activities of extracts were investigated by tyrosinase inhibition, astringent, anti-bacterial and MTT assay for cell viability. In the whitening effect, PA that the highest tyrosinase inhibition activity showed 56% at 10 ppm in ethanol extract. Also water and ethanol extract of RE showed 54%, 68% at 1,000 ppm, respectively, but LE and CA showed lower effect. Astringent effect of water and ethanol extract of PA appeared over 60% at 1,000ppm concentration but other extracts showed no astringent effect. In the anti-bacterial test, water and ethanol extract of PA showed no anti-bacterial effect against all microorganisms. But water and ethanol extract of RE showed anti-bacterial effect on Staphylococcus aureus, ethanol extract of showed on Staphylococcus aureus and Propionibacterium acnes, ethanol extract of CA showed on Candida albicans. The resUlt of stability test showed that the emulsion of containing PA were very stable at various temperature and sun-light test. Viscosity and pH of emulsion did not change. From the results of human patch test to assess the safety of cosmetics containing PA there was no negative reaction on skin was found.

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In vitro Transport of Fexofenadine.HCl in Deformable Liposomes Across the Human Nasal Epithelial Cell Monolayers

  • Lin, Hong-Xia;Lee, Chi-Ho;Shim, Chang-Koo;Chung, Suk-Jae;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.483-489
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    • 2004
  • Fexofenadine HCl is non-sedating histamine H1 receptor antagonist that can be used for the treatment of seasonal allergic rhinitis. The objective of this study was to investigate whether the carriers of deformable liposomes can enhance the transepithelial permeability of fexofenadine HCl across the in vitro ALI human nasal monolayer model. Characterization of this model was achieved by bioelectric measurements and morphological studies. The passage 2 and 3 of cell monolayers exhibited the TEER value of $2852\;{\pm}\;482\;ohm\;{\times}\;cm^2$ on 11 days of seeding and maintained high TEER value for 5 days. The deformable liposome of fexofenadine HCl was prepared with phosphatidylcholine (PC) and cholic acid using extruder method. The mean particle size was about 200 nm and the maximum entrapment efficiency of 33.0% was obtained in the formulation of 1% PC and $100\;{\mu}g/ml$ fexofenadine HCl. The toxicity of the deformable liposome to human nasal monolayers was evaluated by MTT assay and TEER value change. MTT assay showed that it has no toxic effect on the nasal epithelial cells in 2-hour incubation when the PC concentration was below 1%. However, deformable liposome could not enhance the transepithelial permeability $(P_{app})$ and cellular uptake of fexofenadine HCl. In conclusion, the in vitro model could be used in nasal drug transport studies and evaluation of transepithelial permeability of formulations.

Effects of Chitosan on the Cytotoxicity of Anticancer Drugs in vitro (In vitro에서 chitosan이 항암제의 세포독성에 미치는 영향)

  • Min, Soon-Hong;Pyo, Myoung-Yun
    • Environmental Analysis Health and Toxicology
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    • v.22 no.3
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    • pp.263-269
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    • 2007
  • Chitosan is a depolymerized and partially deacetylated derivative of chitin. We investigated the cytotoxicity of chitosan in cancer cell lines, such as P388, L1210, HCT-15, SK-HepG-1 and mouse splenocytes as a normal cell by MTT assay. To clarify whether chitosan enhances cytotoxicity of anticancer drugs, we also examined the cytotoxicity of combined treatment with chitosan and anticancer drugs, such as cisplatin, mitomycin C, and 5-fluorouracil in cancer cell lines in vitro. Chitosan ($37.5\;{\mu}g/mL,\;75\;{\mu}g/mL,\;112.5\;{\mu}g/mL,\;and\;150\;{\mu}g/mL$) showed concentration-dependent cytotoxicity in the cancer cell lines. In addition, chitosan showed relatively lower cytotoxicity in normal cells than in the cancer cell lines. Particularly, this trend was significant at high doses of chitosan, i.e. $112.5\;{\mu}g/mL,\;and\;150\;{\mu}g/mL$. Thus, these results suggest that chitosan may selectively induce the growth inhibition in cancer cell lines, compared to normal cells. Furthermore. the co-treatment of chitosan and anticancer drugs exhibited an apparant synergistic cytotoxicity in murine lymphoma cell lines, i.e. P388 and L1210 at $37.5\;{\mu}g/mL$ of chitosan rather than at $75\;{\mu}g/mL$ of chitosan, but such phenomenon could not be observed in solid tumor cell lines, i.e. HCT-15 and SK-HepG-1. However, chitosan did'nt reduced the cytotoxicity against normal mouse splenocytes induced by anticancer drugs. Therefore, it is concluded that the combination of chitosan and anticancer drugs might be useful for the cancer chemotherapy.

Biocompatibility of two experimental scaffolds for regenerative endodontics

  • Leong, Dephne Jack Xin;Setzer, Frank C.;Trope, Martin;Karabucak, Bekir
    • Restorative Dentistry and Endodontics
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    • v.41 no.2
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    • pp.98-105
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    • 2016
  • Objectives: The biocompatibility of two experimental scaffolds for potential use in revascularization or pulp regeneration was evaluated. Materials and Methods: One resilient lyophilized collagen scaffold (COLL), releasing metronidazole and clindamycin, was compared to an experimental injectable poly(lactic-co-glycolic) acid scaffold (PLGA), releasing clindamycin. Human dental pulp stem cells (hDPSCs) were seeded at densities of $1.0{\times}10^4$, $2.5{\times}10^4$, and $5.0{\times}10^4$. The cells were investigated by light microscopy (cell morphology), MTT assay (cell proliferation) and a cytokine (IL-8) ELISA test (biocompatibility). Results: Under microscope, the morphology of cells coincubated for 7 days with the scaffolds appeared healthy with COLL. Cells in contact with PLGA showed signs of degeneration and apoptosis. MTT assay showed that at $5.0{\times}10^4$ hDPSCs, COLL demonstrated significantly higher cell proliferation rates than cells in media only (control, p < 0.01) or cells co-incubated with PLGA (p < 0.01). In ELISA test, no significant differences were observed between cells with media only and COLL at 1, 3, and 6 days. Cells incubated with PLGA expressed significantly higher IL-8 than the control at all time points (p < 0.01) and compared to COLL after 1 and 3 days (p < 0.01). Conclusions: The COLL showed superior biocompatibility and thus may be suitable for endodontic regeneration purposes.

Cytotoxicity of Angelicae Radix from Korea, China, and Japan on HepG2 Cells (한국 ${\cdot}$ 중국 ${\cdot}$ 일본 당귀(當歸)의 HepG2 세포 독성 비교 연구)

  • Park, Wan-Su;Oh, Myung-Sook;Chang, Mun-Seog;Yang, Woong-Mo;Lee, Byong-Hee;Kim, Won-Nam;Lee, Hak-Chul;Kang, Soon-Ah;Park, Seong-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.5
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    • pp.1155-1158
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    • 2006
  • The purpose of this study is to examine the cytotoxicity of species of Angelica (Angelicas Radix; the root of Angelica gigas Nakai, A. sinensis (Oliv.) Diels, and A. acutiloba Kitag.) on HepG2 cells. The water extracts of roots of Angelica gigas (WAG), A. sinensis (WAS), and A. acutiloba (WAA) were studied for HepG2 cell viability by a modified MTT assay in the concentrations of 5, 10, 50, 100, 250, 500 ug/ml for 24, 48, 72 h. WAG and WAS did not reduced the cell viability significantly. But WAA reduced the cell viability in the concentration of 500 ug/ml for 24 h (85.45%), 48 h (75.01%). In conclusion, WAG and WAS have not the significant cytotoxicity on HepG2 cells in the suitable dose.

Synergistic Effect of Methanol Extract from Kalopanax pictus and Ascorbic Acid on Antioxidant, Anticancer and Immunomodulatory Activities (음나무(Kalopanax pictus) 추출물과 비타민 C의 항산화, 항암 및 면역활성 상승효과)

  • Shon, Mi-Yae
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1634-1640
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    • 2007
  • The 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS), nitric oxide (NO) scavenging activities and ferric- reducing/antioxidant power (FRAP) assay against extracts of Kalopanax pictus (KP) were measured. Radical scavenging and antioxidant activities were increased depend on the concentration and the effects were enhanced by ascorbic acid (AA). KP extracts and AA had a good anti-proliferating activity against HepG2 cells by MTT assay and induces cells apoptosis, which was demonstrated by flow cytometric analysis. KP extracts and AA caused the arrest of cell-cycle progression at either G0-G1-phase or G2/M-phase, which might be depending upon the KP extracts concentration. In addition, KP extracts and AA are effective in enhancing immunity and nitric oxide production by RAW 264.7 macrophages cells. KP extracts and AA inhibited tumor cell growth and exerted antioxidant effects as compared to controls. These results demonstrate that simultaneous AA and KP extracts treatment could be useful in preventing the oxidative damage and anti-proliferating HepG2 cells, and are effective in enhancing immunomodulatory and antioxidant activity.