• Title/Summary/Keyword: cell viability assay

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Effects of Taeyeumjoweetang against Glucose Oxidase-induced Neurotoxicity in the Cultured Mouse Cerebral Cortical Neurons (태음조위탕(太陰調胃湯)이 Glucose Oxidase에 의해 손상된 대뇌피질(大腦皮質) 신경세포(神經細胞)에 미치는 영향(影響))

  • Kim, Jong-kwan;Ryu, Do-gon;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.267-281
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    • 1999
  • 1. Purpose : The purpose of this study was to determine the effects of Taeyeumjoweetang on the cerebral neurons injured by glucose oxidase(GO). 2. Methods : I observed cell viability in mouse cerebral neurons exposed to glucose oxidase by NR assay and MTT assay and determined lipid peroxidation in mouse cerebral neurons exposed to glucose oxidase. After administration of Taeyeumjoweetang water extracts, I observed significant changes of cell viability, lipid peroxidation in mouse cerebral neurons exposed to glucose oxidase. 3. Results : GO induced cell degeneration such as the decrease of cell viability was measured by MTT and NR assay in the cultured mouse cerebral cortical neurons. Taeyeumjoweetang was effective in the increase of total protein of neurons inhibited by GO. Taeyeumjoweetang was effective in the decrease of lipid peroxidation of neurons produced by GO.

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Molecular Biological Study of Anti-cancer Effects of Bee Venom on Human Melanoma Cell (약침용봉독액(藥鍼用蜂毒液)이 흑색종세포(黑色腫細胞)에 미치는 항암효과(抗癌效果)에 대(對)한 분자생물학적(分子生物學的) 연구(硏究))

  • Park, Chan-Yol;Nam, Sang-Soo;Kim, Chang-Hwan;Lee, Jae-Dong;Kang, Sung-Keel;Lee, Yun-Ho;Ahn, Byoung-Choul
    • Journal of Acupuncture Research
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    • v.17 no.2
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    • pp.169-186
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    • 2000
  • To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability, apoptosis, and cell cycle were analyzed using MTT assay, tryphan blue assay, [3H]thymidine release assay, flow cytometric analysis, activity of caspase-3 protease activity assay, and immunocytometric analysis of PCNA. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis- and cell cycle-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, [$^3H$]thymidine release assay, and flow cytometric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and $Bcl-X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment. 5. In flow cytometric analysis of cell cycle and immunocytometric analysis of PCNA expression, cell numbers of $G_1$ phase was increased by a dose-dependant manner. 6. In quantitative RT-PCR analysis of the cell cycle-related genes, p21, p27, and p57 were increased, while Cyclin D1, CDK4, c-Myc, c-Fos, and Histone H3 were decreased. In contrast, there were no remarkable changes in expression levels of CDC2 and c-Jun.

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Evaluation of vitrification for cryopreservation of teeth

  • Dissanayake, Surangi C.;Che, Zhong-Min;Choi, Seong-Ho;Lee, Seung-Jong;Kim, Jin
    • Journal of Periodontal and Implant Science
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    • v.40 no.3
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    • pp.111-118
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    • 2010
  • Purpose: The aim of this study was to investigate whether vitrification in the cryopreservation of periodontal ligament (PDL) cells could be useful for tooth banking. Methods: In step 1, primary cultured human PDL cells were cryopreserved in 100% conventional cryopreservation media and 100% vitrification media (ESF40 media) in different temperatures for 2 weeks. In step 2, a series of modified vitrification formulae named T1 (75% vitrification media + 25% F-media), T2 (50% vitrification media + 50% F-media) and T3 (25% vitrification media + 75% F-media) were used to store PDL cells for 2 weeks and 4 weeks in liquid nitrogen. MTT assay was performed to examine the viability of PDL cells. Results: Maximum cell viability was achieved in cells stored in 100% conventional cryopreservation media at $-196^{\circ}C$ (positive control group) in step 1. Compared to the positive control group, viability of the cells stored in 100% vitrification media was very low as 10% in all test conditions. In step 2, as the percentage of vitrification media decreased, the cell viability increased in cells stored for 2 weeks. In 4-week storage of cells in step 2, higher cell viability was observed in the T2 group than the other vitrification formulae while the positive control group had the highest viability. There was no statistically significant difference in the cell viability of 2-week and 4-week stored cells in the T2 group. Conclusions: These observations indicate 100% vitrification media is not successful in PDL cell cryopreservation. Conventional cryopreservation media is currently the most appropriate media type for this purpose while T2 media would be interesting to test for long-term storage of PDL cells.

Protective Effect of Omega-3 of Polyunsaturated Fatty Acid Docosahexaenoic Acid on the Organic Mercury-Induced Cytotoxicity in Cultured NIH3T3 Fibroblasts

  • Ha, Dae-Ho;Lee, Jai-Kyoo
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.187-192
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    • 2008
  • To clerify the protective effect of omega-3 of polyunsaturated fatty acid docosahexaenoic acid (DHA) on the cytotoxicity induced by organic mercury in cultured NIH3T3 fibroblasts. The measurement of cell viability on ogranic mercury wad done by XTT assay after NIH3T3 fibroblasts were cultured with various concentrations of methyl mercuric chloride (MMC). And also, the effect of DHA on the MMC-mediated cytotoxicity was examined by cell viability, and antioxidant effect of DHA was also assessed by superoxide dismutase (SOD)-like activity and the lipid peroxidation activity in cultured NIH3T3 fibroblasts. In this study, MMC decreased cell viability and $XTT_{50}$ value was determined at $50{\mu}M$ of MMC in these culture. In the effect of DHA against the cytotoxicity induced by MMC, DHA significantly increased the cell viability damaged by MMC in cultured NIH3T3 fibroblasts. And also, DHA showed the antioxidant effect by showing the increase of SOD-like activity and the decrease of lipid peroxidation activity. From these results, it is suggested that organic mercury such as MMC has highly toxic effect on cultured NIH3T3 fibroblasts, and also, omega-3 of polyunsaturated fatty acid, DHA showed the protection on MMC-induced cytotoxicity and antioxidant effect.

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Antioxidant Effect of Poncirin and Cytotoxicity on Cultured Human Skin Fibroblast Damaged by Methyl Mercury

  • Jung, In-Ju;Back, Jong-Cheon;Choi, Yu-Sun
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.355-360
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    • 2007
  • In order to evaluate on the cytotoxicity of methyl mercury (MM) and antioxidant effect of phenolic compound, poncirin against MM-induced cytotoxicity, XTT assay was performed to determine the cell viability after human skin fibroblasts (Detroit 51) were grown in the media containing various concentrations of methylmercuric chloride (MMC). And also, the antioxidant effect of poncirin on the cytotoxicity induced by MMC was examined by cell viability and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in these cultures. MMC decreased cell viability in dose-dependent manner in these cultures and the midcytotoxicity value was determined at concentration of 30 ${\mu}M$ MMC after human skin fibroblasts were treated with $10\sim50{\mu}M$ MMC for 72 hours, respectively. MMC was highly toxic on cultured human skin fibroblasts by toxic criteria. MMC-mediated cytotoxicity was related with oxidative stress by the diminution of toxic effect according to the treatment of vitamin E. In the antioxidant effect of poncirin, it showed vitamin E-like DPPH radical scavenging activity at 90 ${\mu}g/ml$ poncirin and also, remarkably increased cell viability compared with MMC-treated group. From these results, it is suggested that MMC-mediated cytoxicity was highly toxic and was related with oxidative stress in cultured human skin fibroblasts, and also phenolic compound such as poncirin showed the protection on MMC-induced cytotoxicity by antioxidant effect in these cultures.

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Cytotoxicity of Impregnated Dental Gingival Retraction Cord Extracts in Immortalized Human Oral Fibroblasts and Keratinocytes in vitro

  • Myung-Jin Lee;Song-Yi Yang
    • Korean Journal of Materials Research
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    • v.33 no.4
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    • pp.130-134
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    • 2023
  • This study evaluated cell viability and cytokine release in immortalized human oral fibroblasts (hTERT-hNOFs) and keratinocytes (IHOK) exposed to a dental-impregnated gingival retraction cord. To prepare the extracts, dental gingival retraction cords impregnated with aluminum chloride hexahydrate were immersed in a cell culture medium for 24 h at 37 ℃. hTERT-hNOFs and IHOK were cultured for 24 h. The cell culture medium was removed and extracts of the dental gingival retraction cords were added. After incubation with the extract solution, cell viability was evaluated using an MTT assay. The levels of the cytokines IL-1α and IL-8 were measured in the supernatants of each cell type. The cell viability after exposure to the extract solution for 10 min exceeded 70 % in both cell types. The ET50 values for hTERT-hNOF and IHOK were 35.75 and 28.98 min, respectively. For IHOK, the IL-1α level was (5.35 ± 5.22) pg/mL at 10 min, (3.58 ± 5.38) pg/mL at 20 min, and (2.85 ± 4.28) pg/mL at 60 min of exposure (p > 0.05). The IL-8 level in IHOK was (67.16 ± 18.70) pg/mL at 10 min, (78.36 ± 7.50) pg/mL at 20 min, and (111.9 ± 26.10) pg/mL at 60 min of exposure (p > 0.05). Cytokine release was not observed from hTERT-hNOFs. Based on these results, cell viability and cytokine release were confirmed in cells exposed to the impregnated gingival retraction cord. In addition, the application of the extracts to hTERT-hNOF and IHOK during the actual contact time and determination of ET50 may be beneficial for evaluating the biocompatibility of dental-impregnated gingival retraction cords.

Anticancer effect of methanol extract of Erigeron bonariensis on oral cancer cells (구강암 세포주에서 실망초 추출물의 항암효능)

  • Jang, Boon-Sil;Lim, Sun-A
    • Journal of Korean society of Dental Hygiene
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    • v.20 no.5
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    • pp.763-771
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    • 2020
  • Objectives: Erigeron bonariensis is a type of Erigeron found throughout the tropical and subtropical areas as one of the perennial plants or pioneer plants. It is known to show detoxifying, antipyretic, and anticancer effects for various cancers. However, there are no reports on the anticancer effect of E. bonariensis on oral cancer cells. In the present study, we investigated the effects of the methanol extract of Erigeron bonariensis (MEEB) on the inhibition of cell growth and induction of apoptosis in mucoepidermoid carcinoma (MEC) cell lines, including the MC3 and YD15 oral cancer cells. Methods: MC3 Cells were treated by dimethyl sulfoxide (DMSO) or methanol extracts of 20 various natural products 20 ㎍/mL for 48 hours and cell viability were analyzed as Trypan blue exclusion assay. The effects of MEEB treatment on the cell viability of MC3 and YD15 cells, for 48 h, were analyzed by Trypan blue exclusion assay. The anticancer efficacy and apoptosis of oral cancer cell lines were analyzed by western blot analysis. The statistical significance of differences between groups was analyzed by Student's two-tailed t-test. A value of P<0.05 compared to the vehicle control was considered statistically significant. Results: Among 20 different naturally derived products, MEEB significantly inhibited cell viability and increased cleaved poly [ADP-ribose] polymerase 1 (PARP) protein in the MC3 and YD15 cells in a concentration-dependent manner. Conclusions: These results suggest that MEEB can be used as a natural anticancer drug for the treatment of human oral cancer.

Antioxidant Effect of Cynomorii Herba on HepG2 Cells and Diphenyl-picryl-hydrazyl (DPPH) Radical Scavenging Activity (쇄양(鎖陽)의 Diphenyl-picryl-hydrazyl (DPPH) 소거 활성 및 HepG2 세포에 대한 항산화 효과)

  • Chang, Mun-Seog;Yang, Woong-Mo;Kim, Do-Rim;Park, Eun-Hwa;Park, Soo-Yeon;Park, Seong-Kyu
    • Herbal Formula Science
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    • v.15 no.2
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    • pp.139-145
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    • 2007
  • The purpose of this study was to investigate the anti-oxidant effect of Cynomorium songaricum. The extract of Cynomorii Herba was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, HepG2 cell viability and $H_2O_2$-induced cytotoxicity by a modified MTT assay. DPPH radical scavenging activity was measured after 30 minutes. The extract was tested by 1. 5, 10, 50, 100 and 500 ${\mu}g/ml$ concentrations. HepG2 cell viability by a modified MTT assay was measured in the concentrations of 10, 50, 100, 250, 500 ug/ml for 24 h. The results showed that the extract scavenged DPPH radical up to 52.2% with 50 ug/ml concentration. The extract did not reduced the cell viability and $H_2O_2-induced$ cytotoxicity (69.4%) was blocked by the extract in the concentrations of 50, 100, 250 and 500 ${\mu}g/ml$. In conclusion, the extract of Cynomorii Herba has potent antioxidant activity.

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Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method

  • Jang, Sun Mi;Cho, Sin-Yeon;Kim, Eui-Seong;Jung, Il-Young;Lee, Seung Jong
    • Journal of Korean Dental Science
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    • v.9 no.1
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    • pp.1-8
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    • 2016
  • Purpose: The aim of this study was to evaluate the rat periodontal ligament cell viability under $0^{\circ}C/2$ MPa condition up to one week using in vivo 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay. Materials and Methods: As soon as 110 upper molar teeth of rats were extracted, they were stored in Hartman's solution under $0^{\circ}C/2$ MPa condition for 1, 2, 3, 4, and 7 days each. All specimens were treated with in vivo MTT assay and the value of optical density was measured by ELISA reader. These values were statistically analyzed by one-way ANOVA. Result: There was no statistical difference on MTT value between immediate and 1 day storage group. There were statistically significant differences between 1 day and 2 days tsorage, 2 and 3 days storage groups, respectively. Teeth of 34, and 7 days storage groups showed significantly lower MTT valuesc ompared with shorter period storage groups. Conclusion: When the MTT values were substituted in standard curve, 1 day storage group at $0^{\circ}C/2$ MPa condition showed 68% cell viability when compared with immediate group. It dropped to 13% at 2 days, and to less than 5% at 3 days or more.

Applicability of Fluorescein Diacetate (FDA) and Calcein-AM to Determine the Viability of Marine Plankton (FDA와 Calcein-AM 방법을 이용한 해양플랑크톤 생사판별기법)

  • Baek, Seung-Ho;Shin, Kyoung-Soon
    • Ocean and Polar Research
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    • v.31 no.4
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    • pp.349-357
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    • 2009
  • Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.