• Title/Summary/Keyword: MTT 분석법

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Effect of Epimedium Koreanum Nakai on GO-Induced Neurotoxicity in Cultured Mouse Spinal Dorsal Root Ganglion Neurons (Glucose Oxidase에 의(依)하여 손상(損傷)된 배양척수감각신경절세포(培養脊髓感覺神經節細胞)에 대(對)한 음양곽(淫羊藿)의 효과(效果))

  • Park Seung-Taeck;Lee Ho-Sub;Yun Yong-Gap;Park Byung-Rim
    • Herbal Formula Science
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    • v.7 no.1
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    • pp.143-151
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    • 1999
  • To evaluate the neurotoxic effect of oxygen radicals in cultured mouse spinal dorsal root ganglion(DRG) neurons, cytotoxicity was determined by MTT assay after cultured DRG neurons were grown in the media containing various concentrations of glucose oxidase(GO). In addition, neuroprotective effect of herb extract, Epimedium Koreanum Nakai was examined by MTT assay in cultured DRG neurons. Cell viability of cultured DRG neurons was remarkably decreased by GO in dose- and time-dependent manner, and Epimedium Koreanum Nakai protected remarkably GO-induced neurotoxicity in these cultures. From the above results, it is suggested that oxygen radicals is toxic in cultured mouse DRG neurons, and herb extracts such as Epimedium Koreanum Nakai are effective in prevention of the neurotoxicity induced by oxygen radicals in cultured mouse DRG neurons.

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Synthesis and bioactivity evaluation of metal ion-substitution biphasic calcium phosphate for bone defect reconstruction (골결손부 재건을 위한 금속 이온 치환 이상인산칼슘 합성 및 생체 활성 평가)

  • Kim, Tae-Wan;Kim, Dong-Hyun;Jin, Hyeong-Ho;Lee, Seung Ho;Park, Hong-Chae;Yoon, Seog-Young
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.22 no.6
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    • pp.279-285
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    • 2012
  • The co-precipitation technique has been applied to synthesize Biphasic Calcium Phosphate (BCP), Mg-BCP and Si-BCP. X-ray diffraction (XRD) and Fourier transformed infrared (FTIR) spectroscopy were used to characterize the structure of synthesized BCP, Mg-BCP and Si-BCP powders. The results have shown that BCP and substitution of magnesium and silicon in the calcium deficient apatites revealed the formation of biphasic mixtures of Hydroxyapatite (HAp)/${\beta}$-Tricalcium phosphate (${\beta}$-TCP) ratios after heating at $1000^{\circ}C$. Ionic substituted BCP is able to develop a new apatite phase on the surface in contact with physiological fluids faster than BCP does. An MTT assay indicated that BCP, Mg-BCP, and Si-BCP powders had no cytotoxic effects on MG-63 cells, and that they have good biocompatibility.

Effect of Angelica keiskei Extract on Apoptosis of MDA-MB-231 Human Breast Cancer Cells (신선초 추출물이 인체 유방암 세포 MDA-MB-231의 세포 사멸에 미치는 영향)

  • Jeong, Yu-Jin;Kang, Keum-Jee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.12
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    • pp.1654-1661
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    • 2011
  • We investigated the effect of Angelica keiskei ethanol (AKE) extract on cell death in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured in the presence 125, 150 and 175 ${\mu}g$/mL concentrations of AKE for 24 hours. MTT assays demonstrated that mitochondrial dehydrogenase activities decreased in a dose-dependent manner in MDA-MB-231 cells (p<0.05). In contrast, the proportion of dual staining with Hoechst 33342/ethidium bromide(EtBr) for cell death increased in a dose-dependent manner in MDA-MB-231 cells (p<0.05). In particular, the levels of cell death caused by apoptotic program showed marked increases in the 150 and 175 ${\mu}g$/mL AKE groups, as revealed by flow cytometry. An apoptotic suppressor gene, Bcl-2, significantly decreased at the transcript level (p<0.05). The expression levels of proapoptotic genes, both Bax and caspase 3 significantly increased (p<0.05). Furthermore, the ratio of Bcl-2/Bax mRNA which is considered to be an important indicator of apoptosis, significantly decreased in a dose-dependent manner (p<0.05). These results taken together indicate that, the AKE extract used in this study induces cell death in MDA-MB-231 human breast cancer cells.

Preparation and Characterization of PLGA Scaffold Impregnated Keratin for Tissue Engineering Application (케라틴이 함유된 조직공학적 PLGA 지지체의 제조 및 특성 분석)

  • Oh, A-Young;Kim, Soon-Hee;Lee, Sang-Jin;Yoo, James J.;Dyke, Mark van;Rhee, John M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.5
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    • pp.403-408
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    • 2008
  • Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.

Anti-tumor effect of new compound, 127, through the induction of apoptosis (새로운 화합물 c-127의 세포고사 유도에 의한 항암효과)

  • Baek, Ki Hwan;Han, A Lum;Shin, Sae Ron;Jin, Chun Mae;Yoon, Young Wook;Yu, Seung Taek;Kim, Jong Duk;Choi, Du Young
    • Clinical and Experimental Pediatrics
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    • v.52 no.6
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    • pp.696-700
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    • 2009
  • Purpose : We screened more than 350 compounds with an endoperoxide ring structure in search of an anti-leukemic drug and found that compound 127 (c-127) could induce significant cytotoxicity in HL-60 cells. In this study, we investigated the molecular mechanisms of compound 127-induced antitumor activity on HL-60 cells. Methods : HL-60 cells were cultured in Rosewell Park Memorial Institute 1640 and cell viability was measured by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], a tetrazole assay. Apoptosis was assessed by a DNA fragmentation test. Apoptotic machineries were determined by Western blot analysis. Results : C-127 could induce a cytotoxic effect at 24 h and apoptosis at 6 h, which was demonstrated with MTT assay and DNA fragmentation test, respectively. The apoptotic effect of this drug was caused by the activation of the intracellular caspase-8,3 activation, the cleavage of pro-apoptotic Bid, and the increase of c-Jun expression accompanied with JNK (Jun N-terminal kinases) phosphorylation. On the contrary, it increased the expression of anti-apoptotic Bcl-2 levels, leading to the induction of the induction of anti-apoptotic effect. Taken together, the present study demonstrated that c-127 was a potent inducer of cytotoxicity on HL-60 cells through apoptotic mechanisms, which included the activation of caspase family, the regulation of Bcl-2 family, and the activation of JNK signaling pathway. Conclusion : Our results suggest that c-127 has a strong antitumor activity through the regulation of various apoptotic machineries on HL-60 cells. The compound may be utilized as an effective and potentially therapeutic drug in leukemia.

Enhancement of Cytotoxicity by the Combination of Anticancer Drugs in Human Lung Adenocarcinoma Cell Line (PC-14) (폐암세포주 (PC-14)에서 복합항암제 처치시 암세포살해능의 증강에 관한 연구)

  • Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.525-533
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    • 1997
  • Background : No ideal combination chemotherapy for lung cancer has been established even though lots of combination anticancer chemotherapies have been tried. For the combination of anticancer drugs, the interaction of anticancer drugs is very important but unpredictable factor. In this experiment, we designed and tested new experiment to measure the interaction of two anticancer drugs using MIT assay in an attempt to predict clinical response of the combination regimen. Methods : With human lung adenocarcinoma cell line (PC-14), the cytotoxic effect of cisplatin, adriamycin, mitomycin C and etoposide were measured by in vitro chemosensitivity test (MIT assay). The combined cytotoxic effects of combination of two drugs were also measured in every combination of the drug concentrations and analyzed the interaction by Anava analysis of two way factorial design. Results : Four individual drugs showed cytotoxic effects on PC-14 by dose dependent fashion. Comparison of two drug combinations revealed that mitomycin C + cisplatin and adriamycin + cisplatin combinations showed stronger synergistic cytotoxic effects. Conclusion : From this experiment, we suggest two combinations of mitomycin C + cisplatin and adriamycin + cisplatin as chemotherapeutic regimens for unresectable non-small cell lung cancer. Furthermore, this experimental design could be applied to other types of cancer requiring combination anticancer chemotherapy.

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The Effect of Red Cabbage (Brassica oleracea L. var. capitata f. rubra) Extract on the Apoptosis in Human Breast Cancer MDA-MB-231 Cells (적양배추 추출물이 인체 유방암 세포 MDA-MB-231의 세포사멸에 미치는 영향)

  • Nam, Mi Kyung;Kang, Keum Jee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.1
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    • pp.8-16
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    • 2013
  • We investigated the effect of red cabbage extract (RCE) on cell death in MDA-MB-231 human breast cancer cells. Cells were cultured in the presence 1.0, 1.5, and 2.0 mg/mL concentrations of RCE for 24 hours. MTT assays demonstrated that mitochondrial dehydrogenase activities decreased in a dose-dependent manner in cells (p<0.05). In contrast, the proportion of dual staining with Hoechst 33342/ethidium bromide (EtBr) for cell death increased in a dose-dependent manner in cells (p<0.05). Flow cytometry assays revealed that cell death caused by an apoptotic program increased in a dose-dependent (p<0.05). Also, increased ROS accumulation in cells, as revealed by DCF-DA staining, was observed in a dose-dependent fashion (p<0.05). The apoptosis suppressor gene Bcl-2 decreased significantly at the mRNA level. Pro-apoptotic genes Bax and caspase-3, genes that are related to the last stage of apoptosis significantly increased. The Bcl-2/Bax ratio which is an important indicator of apoptosis, was found to have significantly decreased dose dependence. These results taken together indicate that the effect of red cabbage extract induces cell death in MDA-MB-231 human breast cancer cells.

Evaluation of the radiopacity and cytotoxicity of resinous root canal sealers (레진계 근관충전실러의 방사선 불투과성 및 세포 독성에 대한 평가)

  • Kim, Chang-Kyu;Ryu, Hyun-Wook;Chang, Hoon-Sang;Lee, Byung-Do;Min, Kyung-San;Hong, Chan-Ui
    • Restorative Dentistry and Endodontics
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    • v.32 no.5
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    • pp.419-425
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    • 2007
  • The aim of this study was to evaluate the radiopacity and cytotoxicity of three resin-based (AH 26, EZ fill and AD Seal), a zinc oxide-eugenol-based (ZOB Seal), and a calcium hydroxide-based (Sealapex) root canal sealers. Specimens, 10 mm in diameter and 1 mm in thickness, were radiographed simultaneously with an aluminum step wedge using occlusal films, according to ISO 6876/2001 standards. Radiographs were digitized, and the radiopacity of sealers was compared to the different thicknesses of the aluminum step wedge, using the Scion image software. Using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the cytotoxicity of each material was determined in immortalized human periodontal ligament (IPDL) cells. The results demonstrated that EZ fill was the most radiopaque sealer, while Sealapex was the least radiopaque (p < 0.05). AH 26, AD Seal and ZOB Seal presented intermediate radiopacity values. All the materials evaluated, except for Sealapex, presented the minimum radiopacity required by ISO standards. The cell viabilities of resin-based root canal sealers were statistically higher than that of other type of root canal sealers through the all experimental time. Further, EZ fill showed statistically lower cell viability in 24 and 48 hours compared to AD Seal and in 72 hours compared to all other resin-based root canal sealers. However, there was no correlation between the radiopacity and cytotoxicity of three resin-based root canals sealers (p > 0.05). These results indicate that resin-based root canal sealer is more biocompatible and has advantage in terms of radiopacity.

고분자량 Chitooligosaccharides의 in vitro 항종양성

  • 박헌국
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2001.12a
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    • pp.127-127
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    • 2001
  • Chitosan에 Ttichoderma viride 유래의 cellulase를 처리하여 효소분해함으로써 고분자량 chitooligosaccharides를 얻었다. 생산된 올리고당을 HPLC로 분석한 결과 octamer 이상의 것이 전체 올리고당 중 49.3%에 달하는 비교적 고분자 화합물로 구성된 chitooligosaccharides의 혼합물이었다. MTT검색법에 의하여 고분자량 chitooligosaccha- rides의 정상세포주와 암세포주에 대한 세포독성을 실험하였다. 정상세포주인 아프리카 초록원숭이의 신장세포에 대한 $IC_{50}$/(50% 저해농도)값은 1,107.95$\mu$g/ml로 정상세포에 대한 독성은 거의 없었다. 폐암세포주인 A549, 방광암세포주인 J82, 대장암세포주인 SNU-C4, 위암세포인 SNU-1, 유방암세포주인 ZR75-1에 대한 $IC_{50}$/값은 각각 421.06$\mu$g/ml, 417.99$\mu$g/ml, 445.54$\mu$g/ml, 380,65$\mu$g/ml, 460.49$\mu$g/ml로 in vitro 종양세포 증식억제 활성을 나타내었다.

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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.