• Title/Summary/Keyword: MTT-test

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산삼의 배양 및 그 응용에 관한 연구

  • Sin, Mi-Hui
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.151-162
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    • 2001
  • Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extrac. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were patch tested in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynnthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively

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Evaluation of effect of galvanic corrosion between nickel-chromium metal and titanium on ion release and cell toxicity

  • Lee, Jung-Jin;Song, Kwang-Yeob;Ahn, Seung-Geun;Choi, Jung-Yun;Seo, Jae-Min;Park, Ju-Mi
    • The Journal of Advanced Prosthodontics
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    • v.7 no.2
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    • pp.172-177
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    • 2015
  • PURPOSE. The purpose of this study was to evaluate cell toxicity due to ion release caused by galvanic corrosion as a result of contact between base metal and titanium. MATERIALS AND METHODS. It was hypothesized that Nickel (Ni)-Chromium (Cr) alloys with different compositions possess different corrosion resistances when contacted with titanium abutment, and therefore in this study, specimens ($10{\times}10{\times}1.5mm$) were fabricated using commercial pure titanium and 3 different types of Ni-Cr alloys (T3, Tilite, Bella bond plus) commonly used for metal ceramic restorations. The specimens were divided into 6 groups according to the composition of Ni-Cr alloy and contact with titanium. The experimental groups were in direct contact with titanium and the control groups were not. After the samples were immersed in the culture medium - Dulbecco's modified Eagle's medium[DMEM] for 48 hours, the released metal ions were detected using inductively coupled plasma mass spectrometer (ICP-MS) and analyzed by the Kruskal-Wallis and Mann-Whitney test (P<.05). Mouse L-929 fibroblast cells were used for cell toxicity evaluation. The cell toxicity of specimens was measured by the 3-{4,5-dimethylthiazol-2yl}-2,5-diphenyltetrazolium bromide (MTT) test. Results of MTT assay were statistically analyzed by the two-way ANOVA test (P<.05). Post-hoc multiple comparisons were conducted using Tukey's tests. RESULTS. The amount of metal ions released by galvanic corrosion due to contact between the base metal alloy and titanium was increased in all of the specimens. In the cytotoxicity test, the two-way ANOVA showed a significant effect of the alloy type and galvanic corrosion for cytotoxicity (P<.001). The relative cell growth rate (RGR) was decreased further on the groups in contact with titanium (P<.05). CONCLUSION. The release of metal ions was increased by galvanic corrosion due to contact between base metal and titanium, and it can cause adverse effects on the tissue around the implant by inducing cytotoxicity.

Antimicrobial activity and cytotoxicity test of Scrophularia ningpoensis hemsl extracts against Klebsiella pneumoniae

  • Yook, Keun-Dol
    • Journal of the Korea Society of Computer and Information
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    • v.21 no.5
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    • pp.135-139
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    • 2016
  • Scrophularia ningpoensis hemsl has been traditionally used in China and Vietnam for treatment of bacteria, atopy, pimple, tonsillitis, angina and encephalitis for a long time. The main objectives of this study were to evaluate the antibacterial activity of the Scrophularia ningpoensis hemsl extract on biofilm formation of Klebsiella pneumoniae. Antibacterial activity was conducted using disc diffusion assay and minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) were determined using the broth micro dilution method in accordance to Clinical and Laboratory Standards Institute guidelines(CLSI). Furthermore, cytotoxicity on L929 were assessed using animal cell culture for the proliferation test(MTT cell assay) and the biofilm forming capacity of the K. pneumoniae were determined using the colony forming unit (CFU) assay. The extract exhibited considerable antibacterial activity. K. pneumoniae was susceptible to the extract with the MIC and MBC of 0.1875 and $1.5mg/m{\ell}$ respectively. Cytoxicity test in L929 showed no sign of toxicity at the concentration of $0.75mg/m{\ell}$ and at the same concentration the extract caused inhibition of bacterial biofilm formation. The extract of Scrophularia ningpoensis hemsl possesses an in vitro antibacterial antibiofilm activities against K. pneumoniae, with no sign of cytoxicity on L929.

Increased Antimutagenic and Anticancer Activities of Chinese Cabbage Kimchi by Changing Kinds and Levels of Sub-Ingredient (부재료 첨가 배추김치의 항돌연변이 및 항암성 증진효과)

  • 박건영;조은주;이숙희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.4
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    • pp.625-632
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    • 1998
  • To enhance the antimutagenic and anticancer activities of chinese cabbage kimchi, 13 kinds of kimchi, which were different kinds and levels of sub-ingredient added kimchi, were prepared and fermented at 15℃ for 1 day and then at 5℃ up to pH 4.3. The antimutagenic effects of the methanol extracts of the kimchi were studied by using Ames mutagenicity test in Salmonella typhimurium TA100 and SOS chromotest in E. coli PQ37. Among the kimchi samples, high ratio of red pepper powder(7%) and garlic(2.8% or 5.2%) added kimchi, 1% chinese pepper powder added kimchi and organic cultivated chinese cabbage kimchi significantly reduced(p<0.05) the mutagenicity induced by aflatoxin B1(AFB1) in Ames test and SOS response against N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in SOS chromotest, and also the kimchi inhibited more effectively(p<0.05) the survival and growth of AGS human gastric adenocarcinoma cells than the standarized kimchi on the SRB assay, MTT assay and growth inhibition test. These results suggest that the antimutagenic and anticancer activites of kimchi can be increased by the sub-ingredients such as organic cultivated chinese cabbage, red pepper powder, garlic and chinese pepper powder.

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Evaluation of Pharmacological Activities of Ethanol Extracts Prepared from Selected Korean Medicinal Plants

  • Khan, Imran;Eum, IM Zi;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.33 no.6
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    • pp.427-437
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    • 2018
  • In this study, 23 ethanolic extracts from 20 medicinal plants were evaluated for biological activities. Results revealed that of 23 samples, seven samples have demonstrated good antimicrobial activity. Minimum inhibitory concentrations were 0.4-2.0 mg/mL, while minimum bactericidal concentrations were mostly high 0.8-2.0 mg/mL for selected seven samples. Five samples revealed > 70 mg gallic acid equivalent (GAE)/g of total phenolic contents. Among test samples, six samples exhibited > 80% inhibition of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and only two samples exhibited > 80% inhibition of 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals. A total of five test samples revealed Trolox equivalent antioxidant capacity more than $1000{\mu}m/g$. The MTT assay indicated that eight test samples exhibited > 90% viability of murine macrophage cells (RAW 264.7) at $250{\mu}g/mL$ and suppressed iNOS mRNA expression at transcriptional level when stimulated by lipopolysaccharide (LPS). Some medicinal plants revealed promising results, and so they have prospective for further more inclusive studies.

In vitro Skin Irritation Test of Honeypolis using Human Skin Model

  • Woo, SoonOk;Han, Sangmi;Hong, Inpyo;Kim, Sung-kuk
    • Journal of Apiculture
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    • v.33 no.4
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    • pp.277-282
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    • 2018
  • Ethanol extracted propolis (EEP) was mixed with honey (honeypolis) to dissolve well in water and in vitro skin irritation test was conducted. In vitro method is designed to predict and classify the skin irritation potential of a chemical by assessment of its effect on $EpiDerm^{TM}$, a reconstituted three-dimensional human epidermis model. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 60 min exposure period. In this study under the given conditions honeypolis showed no irritant effects. Honeypolis meets acceptance criteria if: mean absolute OD 570 nm of the three negative control tissues is ${\geq}0.8$ and ${\leq}2.8$, mean relative tissue viability of the three positive control tissues is ${\leq}20%$, standard deviation of relative tissue viability obtained from each three concurrently tested tissues is ${\leq}18%$. Honeypolis is therefore classified as "non-irritant" in accordance with UN GHS "No Category".

Effect of Cosmetics Contained Isotonic Water Mimicked Body Fluid on Cell Activities and Skin (생체 모사수 화장품이 세포 활성과 피부에 미치는 효과)

  • Park, Sun Young;Lee, Sung Hoon;Kim, Eun Joo;Choi, So Woong;Kim, Ji Young;Cho, Seong A;Cho, Jun Cheol;Lee, Hae Kwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.2
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    • pp.195-201
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    • 2014
  • Body fluid has been studied for diverse fields like Ringer's solutions, artificial joint fluids, cell growth culture media because it plays a crucial role in controlling body temperature and acts as a solvent for diverse metabolite processes in the body and delivery media of mineral, energy source, hormone, signal and drug from and to cell via blood or lymphatic vessel by osmotic pressure or active uptake. Stratum corneum containing extracellular lipids and NMF (natural moisturizing factor) absorbs atmospheric water residing outside of cells and utilize it to hydrate inside of their own. This process is related to skin barrier function. In this study, we conducted the cell viability test with Cell Bio Fluid $Sync^{TM}$, which mimicks body fluids including amino acids, peptides, and monosaccharides to strengthen skin barrier, and the clinical skin improvement test with cosmetics containing Cell Bio Fluid $Sync^{TM}$. In the cell viability test, HaCaT cell was treated with PBS for 3 hours, followed by the treatment of a cell culture medium (DMEM) and isotonic solution (PBS) and Cell Bio Fluid $Sync^{TM}$ for 3 hours each. Then, MTT assay and image analysis were conducted. In the clinical skin improvement test, twenty-one healthy women participated. Participants applied cosmetics containing Cell Bio Fluid $Sync^{TM}$ on their face for a week and evaluated the skin hydration, skin roughness, brightness and evenness. All measurements were conducted after they washed off their face and took a rest under the constant temperature ($22{\pm}2^{\circ}C$) and constant humidity conditions ($50{\pm}5%$) for 20 minutes. All the data were analyzed by SPSS (version 21) software program. Results showed that Cell Bio Fluid $Sync^{TM}$ improved both the cell viability and in vivo skin conditions such as skin hydration, roughness, brightness and evenness.

Effects of Sagunjatang and Sagunja-tang plus Mylabris phalerata on Human Stomach Cancer Cells (사군자탕(四君子湯) 및 사군자탕가반묘(四君子湯加斑猫)가 위암세포에 미치는 영향)

  • Jung, Woo-Young;Ryu, Bong-Ha;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Ki-Won;Sin, Hyeon-Su
    • The Journal of Internal Korean Medicine
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    • v.22 no.4
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    • pp.579-587
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    • 2001
  • The efficacy of Sagunja-tang and Sagunja-tang plus Mylabris phalerata against the human stomach cancer was examined and molecular biological fight of its actions was studied. In the efficacy test of anti-stomach cancer cells growth using the MTT assay, administration of Sagunja-tang resulted in no significant change of stomach cancer cells growth, with the control group. Administration of Sagunja-tang plus Mylabris phalerata resulted in a decrease of stomach cancer cells growth in proportion to the concentration of mylabris phalerata and time, which was significantly different from the control group(significance recognized when p<0.05). In the test using the apoptosis assay, administration of Sagunja-tang showed an increase in apoptosis of human stomach cancer cells, with no significant difference from the control group. Administrating Sagunja-tang plus Mylabris phalerata showed an increase in apoptosis of stomach cancer cells in proportion to the concentration of mylabris phalerata and time, which was significantly different from the control group(significance recognized when p<0.05). In the test using the quantitative RT-PCR to examine stomach cancer cells growth and revelation of apoptosis related genes, administrating Sagunja-tang plus Mylabris phalerata resulted in a decrease of Bcl-2, an anti-apoptosis gene, in proportiong to concentration. No significant change was examined in the revelation of CDK1, Cdc2, Cyclin D1, PCNA, c-myc, which are genes related to the stomach cancer cells growth, and Bax, Bel-XL, the genes related to apoptosis, and p53. Referring to the results above, Sagunja-tang plus Mylabris phalerata may be considered to have an anti-growth efficacy against human stomach cancer cells, and an inducement efficacy. Therefore, it can be clinically implemented in the human stomach cancer.

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Comparison on Anti-oxidant Effects of Pigment Extracts from Fabaceae 5 species (국내 자생 콩과 5종 염료 추출물의 항산화 효과 비교)

  • Kim, ye-Ran;Chang, Kyung-Soo;Chang, Jeong Hyun
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.1
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    • pp.7-16
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    • 2020
  • The Fabaceae family are being used as traditional medicine. The aim of this study was to compare the antioxidant effects as well as the cell protecting effects of extracts of 5 species (Astragalus membranaceus, Caesalpinia sappan L., Glycyrrhiza uralensis F., Pueraria lobate O., Pterocarpus santalinus L.) in Fabaceae family. The extracts from 5 species were tested by radical scavenging activity test, total phenolic contents and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on human liver carcinoma (HepG2) cell line. Anti-oxidant effects of the extracts (5 mg/mL) from C. sappan was 93.49% by radical scavenging activity test. In addition, A. membranaceus extracts showed a weak radical scavenging activity. Anti-oxidant effects of the extracts (5 mg/mL) from A. membranaceus was 7.83% by radical scavenging activity test. Total phenolic contents of the extracts from C. sappan and A. membranaceus were 310.93 mg GAE/g extract, 15.33 mg GAE/g extract, respectively. Cell protecting effects against H2O2 treatment were observed at 100 ㎍/mL concentration of C. sappan and P. santalinus extracts. These results suggest that C. sappan and P. santalinus might be best anti-oxidant in Fabaceae family.

Effects of Benzalkonium Chloride on the growth and survival of Human corneal epithelial cells (점안액 보존제 성분 Benzalkonium Chloride에 의해 유도된 각막상피세포의 세포고사 유도)

  • Kim, Jai-Min;Lee, Seok-Ju;Seo, Eun-Sun
    • Journal of Korean Ophthalmic Optics Society
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    • v.7 no.2
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    • pp.189-195
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    • 2002
  • The aim of this study was to investigate the action of benzalkonium chloride (BAC) used as a preservative in most ophthalmic topical solutions, on human corneal epithelial (HCE) cells in vitro. HCE cell line was exposed to BAC solutions at various concentrations (0.01%~0.0001%) for 15 minutes followed by 24 hours of cell recovery. Cell viability was assessed using MTT assay and chromatin condensation with a Hoechst 33342 test. The expression of membrane protein Fas and Fas ligand was examined by western blot and immunocytochemistry, and DNA fragmentation was studied by agarose gel electrophoresis. A significant decrease of membrane integrity with chromatin condensation was observed with BAC tested at concentrations of 0.005% and higher. BAC was cytotoxic preservatives in this study. An apoptotic mechanism appeared to be present at low concentrations of BAC, whereas a necrotic process appeared at higher concentrations. A functional Fas-mediated apoptotic pathway is present in cultured HCE cells and can be activated by upregulation of Fas expression with BAC.

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