• Title/Summary/Keyword: MTT-test

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In Vitro Skin Irritation Test of Anti-Inflammatory Drugs (소염진통제 약물에 대한 In vitro 피부자극 시험연구)

  • Lee, Jong-Kwon;Kim, Dai-Byung;Lee, Eun-Hee;Lee, Sun-Hee;Ryu, Seung-Rel;Choi, Ki-Hwan;Kim, Yoon-Jeong;Kim, Pu-Young
    • Toxicological Research
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    • v.14 no.3
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    • pp.315-320
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    • 1998
  • In vitro skin iritation of anti-inflammatory drugs was investigated in terms of the cytotoxicity method to human skin fibroblast cells. Five anti-inflammatory drugs (Diclofenac, Naproxen, Meclofenamic acid, Ibuprofen and Fnoprofen) which are commercially available as oral preparations or injections were tested. The cytotoxicity of 5 chemicals was evaluated by using MTT[tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. NRU (neutral red uptake) assay and Alamar Blue assay after fibroblast cells had been exposed to the chemicals for 24 hours or 489 hours. The $IC_{50}$ values of the chemicals showed the comparative strength of cytotoxicity as following order of Meclofenamic acid>Diclofenac>Fenoprofen>Ibuprofen>Naproxen. The values of $IC_{50}$ determined by Alamar Blue assay were lower than those of MTT and NRU assay. These data suggest Alamar Blue assay can be useful method for assessing in vitro skin irritation potential of anti-inflammatory drugs.

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Properties of Fucoidan as Raw Materials of Water-holding Cream and Cosmetics (후코이단의 보습제 및 화장품재료로서 특성)

  • Cha, Sung Hwan;Lee, Jung Shik;Kim, Young Suk;Kim, Donguk;Moon, Jae Cheon;Park, Kwonpil
    • Korean Chemical Engineering Research
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    • v.48 no.1
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    • pp.27-32
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    • 2010
  • Properties of fucoidan used for functional cosmetics agents were studied. Fucoidan was extracted from Undaria pinnatifida sporophylls. To test possibility of fucoidan as a cosmetics material, water-holding property measurement, DPPH free radical scavenging assay and MTT assay were done. Water-holding property of fucoidan was higher than that of hyaruronic acid, which is known as the one of the best water-holding material. The water-holding strength of fucoidan slightly increase as molecular weight of fucoidan decrease. Fucoidan showed high stability from MTT assay and good anti-oxidation property from DPPH assay. To evaluate the effect of water-holding property and anti-alergy property of fucoidan on the atopic dermatitis(AD), 46 AD patients were treated with fucoidan cream. After 6 weeks treatment, Investigation Global Assessment(IGA) scores decreased from 3.04 to 2.15, that is fucoidan cream had a 39.8% benefit effect on atopic dermatitis.

Methanolic Extract of Plumbago Zeylanica - A Remarkable Antibacterial Agent Against Many Human and Agricultural Pathogens

  • Singh, Mukesh Kumar;Pandey, Ajit;Sawarkar, Hemant;Gupta, Anshita;Gidwani, Bina;Dhongade, Hemant;Tripathi, Dulal Krishna
    • Journal of Pharmacopuncture
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    • v.20 no.1
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    • pp.18-22
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    • 2017
  • Objectives: The current investigation was carried out to determine the cytotoxic and the antimicrobial activities of methanolic extracts of Plumbago zeylanica. Methods: The stems, leaves, and whole plants were air dried and extracted with methanol by using a Soxhlet extractor for 72 hours at $55-60^{\circ}C$. The antimicrobial activities were determined from the zones of inhibition, which were measured by using the agar well diffusion method, and the cytotoxicity assays were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. Results: The methanolic extracts of the stem and the leaves of Plumbago zeylanica were tested against six bacterial species and nine fungal species, and both extracts showed antimicrobial activity in a dose-dependent manner. The leaf extract of Plumbago zeylanica showed maximum antimicrobial activity against both Staphylococcus aureus sub sp aureus and Fusarium oxysporum. The stem extract was found to be more antimicrobial against the Pseudomonas aeruginosa and the Penicillium expansum species. MTT assays were used to test the cytotoxicity of the whole plant extract in the HCT-116 and the K-562 cell lines, and that extract was shown to have weak cytotoxicity in both cell lines. Conclusion: In the present study, the methanolic stem extracts of Plumbago zeylanica were found to possess remarkable antibacterial activities against many human and agricultural pathogens. The extracts were also found to possess significant antifungal activities, but the antifungal activities were less than the antibacterial activities. Finally, the extracts were found to have weak cytotoxicities in the HCT-116 and the K-562 cell lines.

A study on the Effects of Organism Vigor Information Solution to the Growth of Tumor-Derived Animal Cell (생물활성정보수가 종양 유래 동물세포 성장에 미치는 영향에 관한 고찰)

  • Jang, M.Y.;Lee, K.S.;Shin, J.W.;Mun, C.W.
    • Journal of Biomedical Engineering Research
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    • v.32 no.3
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    • pp.185-190
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    • 2011
  • The purpose of this study is to investigate the influence of organism vigor information solution(OVIS) on the animal cell growth and to set up a condition for cell culture. We investigated the reactions on the MG-63 and MCF-7 cell line in mixture culture media with various amount of REVIEW solution, which is an example among various OVIS. DMEM-HG was used as basic media. The concentration range of the mixture was limited from 0% to 15%. MTT assay is used for cell viability test. The cell was incubated for 14days and the MTT assay was performed on day 1, 3, 7, 10 and 14 throughout the experiment. We used the ELISA reader to measure the Optical Density(O.D) at 595 nm wavelength filter. MCF-7 was linearly proliferated according to culture time and concentrations of OVIS. On the other hand, MG-63 cells were measured the highest O.D value at 12%. The growth rates of both cells cultured in mixed culture media with OVIS are much higher than those in only basic media, DMEMHG, after 14days. It was confirmed that cell cultured at OVIS grows rapidly at certain period although cells showed a negative effect in initial stage.

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.

Propugnating Effect of Bark of Rhizophora mucronata Against Different Toxicants Viz Carbon Tetrachloride, Ethanol and Paracetamol on HepG2 Cell Lines

  • Jairaman, Chitra;Yacoob, Syed Ali Mohamed;Venkatraman, Anuradha;Nagarajan, Yogananth;Murugesan, Gnanadesigan
    • Journal of Pharmacopuncture
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    • v.22 no.1
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    • pp.41-48
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    • 2019
  • Objective: The aim of the study was to evaluate the hepatoprotective activity of the bark extract (Ethanol: Water) in the ratio of (3:1) of Rhizophora mucronata (BERM) by intoxicating the $HepG_2$ cell lines with different toxicants viz, $CCL_4$, Ethanol and Paracetamol with different concentrations of the extract were used. The $HepG_2$ cell lines were subjected to MTT Assay for studying the cytotoxicity. Methods: $HepG_2$ cells were plated using 96 well plate in 10% bovine serum, exposed to different toxicants viz, 2% $CCl_4$, 60% Ethanol and 14 mM Paracetamol respectively. The various test concentrations (18.85, 37.5, 75, 150 and $300{\mu}g/ml$) of bark extract of Rhizophora mucronata was added and incubated for 24 hours. Medium was removed after incubation period and 0.5 mg/ml MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was added and again incubated for 4 hours at 37oC. Then MTT was removed the crystals was dissolved in DMSO and absorbance was measured at 570 nm. Results: The result showed that dose dependent increase in percentage of viability at the doses of 18.85, 37.5, 75, 150, $300{\mu}g/ml$. Te results for the $CCl_4$ intoxicated, at $300{\mu}g/ml$ of the concentration of the extract, the % of viable cells was found out to be 99.6%, for Ethanol intoxicated, 97.67%, and Paracetamol induced, 75.37%, IC50 was $21.53{\mu}g/ml$, $12.61{\mu}g/ml$ and $21.42{\mu}g/ml$ respectively. Conclusion: Thus, we conclude that, the extract possesses defensive effect against different toxicants and can be used as an alternate drug for hepatotoxicity.

Toxicological Evaluation of Chitosan Cross-linked Collagen-GAG Matrix (CCGM) In vitro and In vivo (Chitosan Cross-linked Collagen-GAG Matrix(CCGM)의 독성학적 고찰)

  • Lee, Hae-Yul;Kim, Dong-Hwan;Cho, Hyun;Ahn, Byoung-Ok;Kang, Soo-Hyung;Kim, Won-Bae
    • Toxicological Research
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    • v.16 no.1
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    • pp.17-25
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    • 2000
  • Chitosan cross-linked collagen-glycosaminoglyan (CCGM) is an artificial skin substitute made to form a sponge like dimensional matrix. It can be used to facilitate reconstruction of dermal tissue when applied on large wounds such as severe burns. In order to study the toxicological effects of CCGM the cytotoxicity, local irritation and skin sensitization test were carried out according to the standards of ISO 10993. In the cytotoxicity test utilizing LDH and MTT test, both the CCGM and its extract had no toxicity of Balb/c 3T3 cells. The local irritatioin test on rabbit skin demonstrated that CCGM did not promote any harmful when directly applied on skin. In addition, it did not elicit any allergic reaction in the guinea pig maximization test. Based on these results, it is suggested that CCGM is a material without cytotoxicity, local irritation and allergenicity.

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The analysis of ballast abrasion and fracture by Multiple Tie Tamper (장비작업에 의한 도상자갈의 마모.파쇄변화에 관한 분석)

  • Lee, Choon-Kil;Kim, Kwan-Hyung
    • Proceedings of the KSR Conference
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    • 2008.11b
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    • pp.1024-1028
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    • 2008
  • The ballast, one of track components, plays an essential role as intermedium in transmitting train load to subgrade safely, and the deterioration of ballast directly effects the growth of track irregularity. In this study, we determined the main factor of ballast deterioration was miniature of ballast gravel caused MTT(Multiple Tie Tamper) works and accumulated traffic loads. To estimate the deterioration characteristics of ballast, we carried out field test through track construction for test and the model test simulating the actual operation environment, have done a comparative analysis with the sample's result(crushing rate) of high-speed railroad running actually.

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An in vitro study of mesenchymal stem cell proliferation on titanium discs coated with rhTGF-β2/PLGA by electrospray (Electrospray법으로 rhTGF-β2/PLGA 복합체를 코팅한 티타늄에서의 간엽줄기세포 증식에 관한 연구)

  • Kim, Joohyung;Kim, Seong-Kyun;Heo, Seong-Joo;Koak, Jai-Young;Lee, Woo-Sung;Lee, Joo-Hee;Park, Ji-Man
    • The Journal of Korean Academy of Prosthodontics
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    • v.54 no.2
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    • pp.120-125
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    • 2016
  • Purpose: The purpose of this study is to identify the effect of mesenchymal stem cell proliferation on recombinant human transforming growth factor-beta (rhTGF-${\beta}2$) / poly (D,L-lactide-co-glycolide) (PLGA) treated titanium discs by electrospray. Materials and methods: Anodized titanium surface coated with PLGA was used for a control group to compare anodized titanium surface coated with 125 ng/ml and 500 ng/ml rhTGF-${\beta}2$ as test groups. Atomic force microscope (AFM) test was utilized to determine the difference in coating surface roughness, and field-emission scanning electron microscopy (FE-SEM) was taken to visualize even distribution of coating particles on titanium discs. The mesenchymal stem cell proliferation was tested by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl-tetrazolium bromide) assay on 1st, 4th, 7th days. Results: According to AFM results, there was no statistically significant difference in titanium discs treated with PLGA and with rhTGF-${\beta}2$/PLGA (P>.05). MTT assay test results showed that there was statistically significant difference in mesenchymal stem cell proliferation on test groups compared to control groups at 7th day, and cell viability on discs coated with rhTGF-${\beta}2$ was significantly higher than control groups (P<.05). Conclusion: Titanium surface coated with rhTGF-${\beta}2$/PLGA shows statistically significant higher cell proliferation and the titanium surface coated with the higher concentration of rhTGF-${\beta}2$ presents faster cell growth activity.

Verification of Cytotoxicity Against Cancer Cell Line and Estrogen-like Activity of Cheongkukjang (청국장의 암세포주에 대한 세포독성 및 에스트로겐 유사활성 검증)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.1
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    • pp.153-157
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    • 2007
  • In order to evaluate the cancer preventive and estrogenic compounds in soybean and Cheongkukjang, MTT assay and in vitro test system for the evaluation of the estrogenic activity were applied. The fractions from the ethanol extract of soybean and Cheongkukjang were prepared by the systematic extraction procedure with the solvents such as hexane, ethyl ether, butanol, methanol and H$_2$O. Ethyl ether fractions of soybean and Cheongkukjang showed the highest cytotoxicity against U937 cell line in dose dependent manner, and ethyl ether fraction of Cheongkukjang showed two times higher cytotoxicity than that of soybean. Aqueous fraction of soybean and ethyl ether fraction of Cheongkukjang revealed the highest estrogenic activity and activity was higher in the fractions of Cheongkukjang than soybean. Mixture of Spirulina and Cheongkukjang showed synergistic activity. These observations concerning cancer preventive and estrogen effects of soybean and Cheongkukjang suggest that these materials possess useful ingredients for the prevention of cancer and/or postmenopausal disorder.