• Title/Summary/Keyword: MTT 실험

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Cytotoxic Effect of Inonotus obliquus Composition in HCT-15 Human Colon Cancer Cells and AGS Gastric Cancer Cells (대장암 세포암종 HCT-15 세포 및 위암 세포암종 AGS 세포에서 차가버섯 조성물에 의한 세포생육 억제 효과)

  • 차재영;전병삼;문재철;유지현;조영수
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.4
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    • pp.633-640
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    • 2004
  • This study was performed to investigate the cytotoxic effect of the water-extract from Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea in HCT-15 human colon carcinoma, AGS human gastric carcinoma and NIH3T3 mouse normal fibroblast cells using viable cell count and MTT assay. The water-extract from Chaga mushroom compositions induced inhibitory effects on proliferation of HCT-15 and AGS cells in the MTT assay and viable cell count. However, mouse normal NIH3T3 cells were exhibited 80% survival under the same condition. Chaga mushroom compositions showed highly antiproliferative effect in human cancer cell line HCT-15 and AGS, but not in mouse normal cell line NIH3T3. These results suggest that Chaga mushroom (Inonotus obliquus) compositions containing powdered green tea are the candidate for chemoprevention in colon and gastric cancer.

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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Preparation and Characterization of Tissue Engineered Scaffold Using Porcine Small Intestinal Submucosa and Hyaluronic Acid (돼지의 소장점막하 조직과 히알루론산을 이용한 조직공학적 담체의 제조 및 특성분석)

  • Lim, Ji-Ye;Kim, Soon-Hee;Kang, Gil-Son;Rhee, John M.
    • Polymer(Korea)
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    • v.32 no.5
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    • pp.415-420
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    • 2008
  • The porcine small intestinal submucosa (SIS) without immunorejection responses and hyalunonic acid (HA) can be used as biomaterials. In this study, we tried to design and characterize novel sponge. SIS- HA sponge was prepared by freeze-drying after addition 1wt% HA solution into fabricated SIS sponge. Sponge was crosslinked with 1-ethyl-(3-3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) solution with 100mM concentration for 24 hrs and lyophilized. SIS-HA sponge was characterized by scanning electron microscopy and fourier transform infrared spectrometer. And water absorption ability of sponge was evaluated. We seeded NIH/3T3 cells in SIS-HA sponge and cellular attachment was assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltertazolium-bromide (MTT) test. We demonstrated presence of HA in SIS-HA sponge from C-O functional group observed by the FT-IR analysis. Moreover, we confirmed low cytotoxicity and high cell viability of the SIS-HA sponges. Therefore, we could expect that SIS- HA scaffolds are applicable for the tissue regeneration.

3-Amino Propane Phosphoric Acid (3-APPA) : A Novel Anti-Aging Substance (3-Amino Propane Phosphoric Acid (3-APPA) : 새로운 피부 노화 억제 물질)

  • 조윤기;변영훈;선보경;황재성;이보섭;김종일
    • Proceedings of the SCSK Conference
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    • 1996.07a
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    • pp.52-67
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    • 1996
  • 새로운 노화방지 물질로 개발한 3-APPA가 노화에 의해 야기되는 여러 변화들, 특히 세포 증식, 유전자 수준 및 단백질 수준에서의 collagen의 생합성 변화, 면역조직화학염색을 이용한 collagen 생합성의 변화등을 세포배양 및 동물실험을 통하여 측정하였다. MTT assay를 이용한 인체 피부 섬유아세포의 증식 실험에서 3-APPA는 무처치군에 비교해서 최고 2배의 섬유아세포 증식 효능을 나타내었으며, $^3$[H]-proline incorporation 방법을 이용한 단층세포 배양 및 3차원 dermal equivalent 섬유아세포 배양에서 무처치군 및 vitamin C 처리군에 비해 최고 1.5배의 collagen 생합성 증가를 나타내었다. 그러나 type I alpha-procollagen mRNA expression에는 영향을 미치지 않는 것으로 나타났다. H&E 염색을 이용한 hairless mice의 피부에 대한 형태학적 변화 및 type I pM procollagen antibody를 이용한 면역조직화학염색에서, 3-APPA는 collagen 생합성을 증가시키는 것으로 나타났다. 이상의 결과에서 3-APPA는 섬유아세포 배양 및 hairless mouse를 이용한 실험에서 피부 섬유아세포 증식을 촉진시키며 collagen 생합성을 증가시켜 피부노화를 억제 할 수 있는 물질임을 밝혔다.

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식분탕(息賁湯)이 항암(抗癌) 및 면역조절작용(免疫調節作用)에 미치는 영향(影響)

  • Yun, Seong-Muk;Ha, Ji-Yong
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.2 no.1
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    • pp.25-42
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    • 1996
  • In order to study the effect of Sikbuntang on the in vitro Cell-cytotoxicity, this study had put through MTT Assay. And to investigate the effects of Sikbuntang on the ICR mice which had Abdominal tumor induced by Sarcoma-180 cell line, C57Bl/6 mice which had pulmonary melanoma induced by B16 cell line. After Sarcoma-180 cell line and B16 cell line were transplanted, the extract of Sikbuntang was orally administered to the mice to observe the extension of survival time of the mice, inhibition of solid tumor, inhibition of pulmonary melanoma metastasis, productivity of Interleukin-2, NK-Activity. The results were summarized as follows : 1. On the MTT assay, in case of $100{\mu}g/ml$ and $10{\mu}g/ml$ of Sikbuntang concentration were inhibited cell viability significantly. But $1{\mu}g/ml$ of Sikbuntang concentration just had tend to inhibit cell viability. 2. In the effect of life extension, Sikbuntang treated group appeared to survive longer than the control group, but which were not significant. 3. In the effect of inhibition solid tumor, Sikbuntang treated group appeared to decrease than the control group, but which were not significant. 4. In the effect of inhibition melanoma pulmonary metastasis, Sikbuntang treated group appeared to inhibit than the control group significantly. 5. In the productivity of Interleukin-2, on 7 and 14 day, Sikbuntang treated group increased than control group significantly. But on 21 day, Sikbuntang treated group had tend to increase with no significance. 6. In the NK-Activity, the ratio of effector cell and target cell. In case which the ratio was 50:1, Sikbuntang treated group showed increase than control group significantly. But in another cases, they showed increase with no significance.

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Studies on the Anticancer Effect of Broussonetia kazinoki Extracts (닥나무(Broussonetia kazinoki) 추출물의 항암효과에 관한 연구)

  • 민경진;정승희;구성자
    • Korean journal of food and cookery science
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    • v.15 no.3
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    • pp.231-237
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    • 1999
  • The anticancer effect of the bark of Broussonetia kazinoki root extracts (hexane. chloroform, ethylacetate, butanol, aqueous) were studied. The cytotoxicity by MTT assay and inhibitory effect on the growth of sarcoma 180 cells were tested in vitro. The reduction rate of the tumor formation and spleen/body weight rate on BALB/c mouse were tested in vivo. From the tests, each fraction showed the cytotoxic effect against the sarcoma 180 cells. In addition, as the concentration of the fractions increased, cytotoxic effect tendency increased as well. The cytotoxic rate of the hexane, chloroform, ethylacetate, butanol and aqueous fractions showed by 58.7%, 40.1%, 75.7%, 52.6% and 62.7% respectively after testing by MTT assay system. And sarcoma 180 cells were incubated for 6 days at 37$^{\circ}C$ with various concentrations of each fraction. As the incubation days go on, the number of cells increased, while the inhibition rate on the growth of sarcoma 180 cells were decreased. Especially the ethylacetate fraction at the concentration of 1.0 mg/ml strongly inhibited the growth of sarcoma 180 cells by 74% compared with the control for a day 37$^{\circ}C$ The hexane, chloroform, ethylacetate, butanol and aqueous fractions inhibited on the growth of sarcoma 180 cells by 31%, 19%, 60%, 30% and 42% respectively, when sarcoma 180 cells has been incubated for 6 days at 37$^{\circ}C$. The each fraction exhibited the antitumor effect in vivo. The ethylacetate fraction reduced the tumor formation by 41% compared with the control, when sarcoma 180 cells were injected subcutaneously into the left groin of BALB/c mice. Also spleen/body weight rate of ethylacetate fraction was increased by 2.10% compared with the control (1.08%). And it is considered that there would be no toxic effect caused by each fraction of body weight and organ as there was on more changes in mouse' weight compared with the control.

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A study of mesenchymal stem cell proliferation and surface characteristics of the titanium discs coated with MS275/PLGA by an electrospray (Electrospray법을 이용한 MS275/PLGA submicron 코팅 티타늄에서의 표면변화와 간엽줄기세포증식에 관한 연구)

  • Yoo, Soo-Yeon;Kim, Seong-Kyun;Heo, Seong-Joo;Koak, Jai-Young;Lee, Joo-Hee;Park, Yoon-Kyung;Kim, Ena
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.4
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    • pp.285-291
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    • 2012
  • Purpose: This study was conducted to identify the surface characteristics of titanium discs coated with MS275/PLGA by electrospray and which is effective to mesenchymal stem cell proliferation. Materials and methods: We used anodized surface coated with PLGA as a control group and anodized surface coated with MS275 $0.5{\mu}M$, $1{\mu}M$, $1.5{\mu}M$ as test groups. To examine that the coating particles are nanometer sized, FE-SEM was used and AFM was utilized to determine the difference of coating surface roughness. We checked the mesenchymal stem cell proliferation by using MTT assay on $1^{st}$, $4^{th}$, $7^{th}$ days. Results: There was no significant difference between control groups and test groups in AFM results (P>.05). In MTT assay results, mesenchymal stem cell proliferation was increased with time, at $7^{th}$ day, cell viability on discs coated with $1.5{\mu}M$ MS275 was significantly higher than control group (P<.05). As SEM showed, the number of cells on all discs was increased and the morphology of cell attachment was also wider and closer with time. Conclusion: Titanium surface coated with MS275/PLGA showed significantly higher cell proliferation and the more density of MS275 was dispersed on titanium discs, the faster cells grew.

Anti-cancer Activity of Supernatant of Rahnella aquatilis AY 2000 Cocultured with Streptomyces griseus (Rahnella aquatilis AY 2000과 Streptomyces griseus의 공배양 상등액의 항암활성)

  • Kim, Ji-Hyun;Kim, Kwang-Hyeon;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.19 no.5
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    • pp.676-679
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    • 2009
  • In order to produce a new antibiotic material against Jurkat T cells using horizontal gene transfer among microbes, co-cultures between soil bacteria AY2000 and the multiple antibiotic producer S. griseus was carried out. It showed that the highest active substance against Jurkat T cells was produced at 48 hr of co-culture time with MTT assay. Moreover, a morphological change of nuclear of Jurkat T cells treated with co-cultured substance was observed in DAPI staining. This result suggests that a new material was produced with co-culture supernatant, and that co-culture between microboes can develop new antibiotic materials.

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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