• Title/Summary/Keyword: XTT Assay

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The Inhibitory Effects of Poncirin against Nickel Induced Cytotoxicity (Poncirin의 니켈에 대한 세포독성억제효과)

  • Yang, Seung-Jin;Kwak, Dong-Keun;Han, Du-Suk
    • The Korea Journal of Herbology
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    • v.21 no.2
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    • pp.121-127
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    • 2006
  • Objectives : Nickel is a major metal used in the nickel-chromium alloys of most orthodontic appliances, partial denture and implants. This study was carried out for the examination of the cytotoxicity on nickel sulfide in cultured NIH3T3 fibroblasts, and poncirin effect on nickel-induced cytotoxicity. Methods : Cell viability for the MTT assay and cell adhesion activity for the XTT assay. Results : The $IC_{50}$ of nickel sulfide by the MTT assay was $93.7\;{\mu}M$. Poncirin was significantly increased the cell viability and cell adhesion activity. Conclusion : Nickel was highly toxic and poncirin has the inhibitory effects against nickel induced cytotoxicity.

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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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Effect of Ziziphi Jujubae Semen on 5-Fluorouracil Induced cytotoxicity in Cultured Vestibular Neurons (배양전정신경세포에 있어서 5-Fluorouracil의 세포독성에 대한 산조인의 효과)

  • Son Il Hong;Lee Jung Hun;Choi Yu Sun;Lee Jae Kyoo;Kim Hyung Su;Lee Yong Suk;Lee Whan Bong;Choi Ki Wook;Min Bu Ki;Kim Sang Su;Lee Kang Chang;Ryu Myeung Hwan;Song Ho Joon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.1
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    • pp.146-149
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    • 2002
  • To evaluate the protective effect of Ziziphi Jujubae Semen(ZJS) on 5-Fluorouracil(5-Fu) in cultured vestibular neurons(VN), neurotoxicity was assessed by XTT assay after VN was exposed to 3-24ug/ml 5-Fu for 48 hours. and also, the neuroprotective effect of ZJS was measured by XTT assay in these cultrures. Cell viability was remarkably decreased dose-dependently, after the treatment with 12ug/ml 5-Fu to cultured VN for 48 hours. In the neuroprotective effect of ZJS on the toxicity induced by 5-Fu, ZJS prevented the neurotoxicity induced by 5-Fu in these cultures. From above the results, it suggests that 5-Fu is toxic in cultured VN and herb extract, ZJS has protective effect over the neurotoxicity induced by 5-Fu.

Toxic Assessment on Effluents of Argo/Industrial Wastewater Treatment Plants in Jeonnam using Chemical and Biological Method (전남 농공단지 폐수처리장 방류수의 화학적/생물학적 독성평가)

  • Lee, Moon-Hee;Choi, Ik-Chang;Han, Sang-Kuk
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.14 no.4
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    • pp.267-273
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    • 2008
  • The purpose of this study is to investigate the distributive property of organic pollutants in effluents of argo/industrial wastewater treatment plants in Jeonnam using simultaneous analysis method of 310 chemicals. The numerous organic pollutants were detected in four sampling sites and the major chemicals were pesticides, CH type chemicals such as polycyclic compounds, CHO type chemicals such as phthalates, and CHO(N) type chemical such as aromatic amines. Moreover, 17 kinds of endocrine disrupters which include diethylptbalate were detected in each sampling sites. TU (Toxic unit) indicated cytotoxicity of samples using XTT assay, it appeared highly at A point(27.2) and D point(24.4). Also, the results from the correlation between total concentration of pollutants and TU did not correspond to the results of chemical and biological analysis.

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Effect of aging on tear strength and cytotoxicity of soft denture lining materials; in vitro

  • Landayan, Jordi Izzard Andaya;Manaloto, Adrian Carlos Francisco;Lee, Jeong-Yol;Shin, Sang-Wan
    • The Journal of Advanced Prosthodontics
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    • v.6 no.2
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    • pp.115-120
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    • 2014
  • PURPOSE. The aim of this in vitro study was to evaluate the effect of aging on the tear strength and cytotoxicity of four soft denture lining materials. MATERIALS AND METHODS. Four commonly used soft denture lining materials, (Coe-Comfort$^{TM}$ GC America Inc., Alsip, IL, USA; Coe-SOFT$^{TM}$ GC America Inc., Alsip, IL, USA; Visco-gel Dentsply Caulk Milford, DE, USA; and Sofreliner Tough M Tokuyama Dental Corporation Tokyo, Japan) were selected. Sixty trouser-leg designed specimens per lining material were fabricated using a stainless steel mold for tear strength testing. The specimens were divided into non-thermocycling and 1000-, and 3000-thermocycling groups. For the cytotoxicity test, twenty-four disk shaped specimens per material were fabricated using a stainless steel mold. The specimens were soaked in normal saline solution for 24 h, 48 h and 72 h. Cytotoxicity was measured by XTT assay in L929 mouse fibroblasts. Data were analyzed by two way analysis of variance and Dunnett's test (P<.05). RESULTS. Before thermocycling, Sofreliner Tough M ($10.36{\pm}1.00N$) had the highest tear strength value while Coe-Comfort$^{TM}$ ($0.46{\pm}0.10N$) had the lowest. After 3000 cycles, Sofreliner Tough M ($9.65{\pm}1.66N$) presented the highest value and Coe-Comfort$^{TM}$ ($0.42{\pm}0.08N$) the lowest. Sofreliner Tough M, in all incubation periods was the least toxic with significant differences compared to all other materials (P<.05). Coe-Comfort$^{TM}$, Coe-$SOFT^{TM}$, and Sofreliner Tough M did not show any significant differences within their material group for all incubation periods. CONCLUSION. This in vitro study revealed that aging can affect both the tear strength and cytotoxicity of soft denture materials depending on the composition.

Effects of Lithospermum erythrorhizon on the cytokine gene expression in human keratinocytes (자초(紫草)가 HaCaT 세포의 사이토카인 유전자 발현에 미치는 영향)

  • Kang, Sang-Hoon;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.50-62
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    • 2013
  • Objective : Propionibacterium acnes (P. acnes) is a major pathogenic bacteria for acne vulgaris. This study was performed to evaluate the effects of Lithospermum erythrorhizon extracts on the inflammatory cytokines gene expression by P. acnes in human keratinocytes, HaCaT cell line. Methods : Anti-bacterial activity and cytotoxicity of LE extracts was analyzed by agar plate culture and XTT assay. The cytokines gene expressions were assessed by real time RT-PCR for IL-8, MCP-1 and TNF-${\alpha}$. During the cell culture and treatments, amounts of secreted TNF-${\alpha}$ were measured by ELISA. Translocation of transcription factor NF-${\kappa}B$ from cytoplasm into nucleus was observed by immunocytochemistry and confocal microscopy. Results : There were no anti-bacterial effects and cytotoxicity as high as $1,000{\mu}g/ml$ of LE extracts in XTT assay. Transcription levels of inflammatory cytokines, IL-8, MCP-1 and TNF-${\alpha}$ were increased by P. acnes in HaCaT. LE extracts decreased the upregulated gene transcription levels. However, amounts of secreted TNF-${\alpha}$ were similar in HaCaT cells with P. acnes and LE extracts. Translocation of NF-${\kappa}B$ into nucleus by P. acnes was significantly inhibited by LE extracts. Conclusions : From the results of this study, LE extracts have anti-inflammatory effects on HaCaT cells by P. acnes that decreased the mRNA expressions of IL-8, MCP-1 and TNF-${\alpha}$. This anti-inflammatory effects of LE extracts could provide the potential of therapeutic substance for acne vulgaris.

Effect of Superoxide Dismutase on Oxidative Stress of Reactive Oxygen Species in Cultured Human Skin Melanocyte (Superoxide Dismutase가 배양인체피부멜라닌세포의 산화적 스트레스에 미치는 영향)

  • Seo, Young Mi;Kim, Nam Song
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.19 no.3
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    • pp.261-269
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    • 2009
  • To evaluate the effect of antioxidant on the cytotoxicity induced by oxidative stress of reactive oxygen species (ROS) in cultured human skin melanocytes, colorimeric assay of XTT and tyrosinase activity assay were adopted after human skin melanocytes were preincubated for 2 hours in the media containing various concentrations of superoxide dismutase (SOD) before the treatment of hydrogen peroxide. Light microscopic study was carried out in same cultures. The results of this study were as follows 1. Cell viability of human skin melanocytes was significantly decreased by 30 and $40{\mu}M$ of hydrogen peroxide($H_2O_2$), respectively. 2. XTT50 was determined at $30{\mu}M$ after human skin melanocytes were treated with $10{\sim}40{\mu}M$ of hydrogen peroxide for 6 hours. 3. The cell viability of cultured human skin melanocytes pretreated with SOD was increased than that of cultured human skin melanocytes treated with $H_2O_2$ dose-dependently. 4. In tyrosinase activity of human skin melanocytes, the cell treated with SOD showed brown stain compared with $H_2O_2$ treated cells, dark stain. 5. In light microscopy, cultured human skin melanocytes exposed to $H_2O_2$ showed morphological changes such as the decreased cell number and cytoplasmic processes, compared with control. 6. In light microscopy, cultured human skin melanocytes pretreated with SOD showed the increase of cell number and cytoplasmic processes compared with $H_2O_2-treated$ group. From these results, it is suggested that oxidative stress of ROS such as $H_2O_2$ has cytotoxicity by showing the decreased cell viability, the increased tyrosinase activity and mophological changes of the decreased cell number and cytoplasmic processes. While, antioxidant like SOD was effective in the prevention of oxidative stress-mediated cytotoxicity by the increased cell viability, decreased tyrosinase activity and the protection of degenerative morphological changes in cultured human skin melanocytes.

Effects of Vaniltic Acid on the Cell Viability and Melanogenesis in Cultured Human Skin Melanoma Cells Damaged by ROS-Induced Cytotoxicity

  • Ha, Dae-Ho;Choi, Yong-Ja;Yoo, Sun-Mi
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.349-354
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    • 2007
  • The purpose of this study was to examine the effect of vanillic acid on the cell viability and melanogenesis in melanocytes damaged by reactive oxygen species (ROS). The human skin melanoma cells (SK-MEL-3) were cultured with various concentrations of hydrogen peroxide $(H_2O_2)$. The cell viability for $H_2O_2$-induced cytotoxicity or vanillic acid against $H_2O_2$ was measured by XTT assay in these cultures. For the effect of vanillic acid on the melanogenesis, the tyrosinase inhibitory activity was measured by colorimetric assay at a wavelength of 490 nm, and melanin synthesis activity were assessed after cells were cultured in the media with or without various cencentrations of vanillic acid. In this study, $H_2O_2$ decreased cell viability dose- and time-dependent manners and $XTT_{50}$ was determined at a concentration of 80 ${\mu}M$, $H_2O_2$. Vanillic acid increased the cell viability dose dependently in human skin melanoma cells damaged by $H_2O_2$-induced cytotoxicity. In the tyrosinase inhibitory activity, vanillic acid supresssed tyrosinase activity in dosedependent manner, and also decreased significantly melanin synthesis activity compared with $H_2O_2$-treated group. From these results. It is suggested that $H_2O_2$-mediated cytotoxicity was highly by the toxic criteria of Borenfreund and Puerner and also, vanillic acid has the protective effect on ROS-induced cytotoxicity and melanogenesis in these cultures.

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Induction of Apoptosis by Realgar on Lung Cancer Cells(A549), Stomach Center Cells(KATO) and Neuroglioma Cells(SNU-1118, U-87MG, U-373MG) (시험관내 폐암(肺癌), 위암(胃癌) 및 신경교종(神經膠腫) 세포(細胞)에 대한 석웅황(石雄黃)의 항암효과(抗癌效果))

  • Bang, Dae-Geon;Kim, Jin-Sung;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.28 no.2
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    • pp.294-303
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    • 2007
  • Objectives : We are aimed to identify anti-tumor effects of realgar on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 5 kinds of cancer cell lines: lung cancer cells(A549). stomach cancer cells(KATO) and neuroglioma cells(SUN-1118. U-87MG, U-373MG). We injected the boiled extracts of realgar $50{\mu}g$. $100{\mu}g$ to cultural media( ml )for 24 hours. We measured the killing effects on 5 kinds of cancer cells through inverted and fluorescence microscope, the suppressive effects on viability of those cells via XTT assay and the effects on the revelation of Bax and Bcl-2 proteins related to apoptosis by western blotting. Results : In the changes of morphology, the extracts of realgar showed more significant killing effects on all cancer cells. especially KATO, SNU-1118, U-87MG, U-373MG, than the control group with dose dependence, which was statistically significant. In XTT assay, the extracts of realgar showed more suppressive effects on viability of all cancer cells, especially KATO and U-373MG, than the control group with dose dependence, which was statistically significant. In the revelation of proteins related to apoptosis, the extracts of realgar increased the level of Bax and decreased that of Bcl-2 in all cancer cells with dose dependence. Conclusions : We identified that realgar had more anti-tumor effects on stomach cancer and neuroglioma than on lung cancer in the experiments above. However, these basic experiments were performed in vitro. We hope the anti-tumor effects of realgar will be practically identified through more progressive research.

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MICROPATTERNED GROOVES AND ACID-ETCHING ON TITANIUM SUBSTRATA ALTER VIABILITY AND GENE EXPRESSION OF ADHERED HUMAN GINGIVAL FIBROBLASTS: A PILOT STUDY

  • Lee, Suk-Won;Kim, Su-Yeon;Lee, Keun-Woo
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.3
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    • pp.375-381
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    • 2007
  • Statement of problem. Prior to determining an optimal width of micropatterned grooves provided on titanium substrata, we have done a pilot study using surface topographies in combined microm and submicrom levels. Purpose. The purpose of this study was twofold 1) to assess the proliferation and 2) to analyze the expression of genes encoding the intracellular signaling proteins involved in cell-substratum adhesions and adhesion-dependent G1 phase cell cycle progression of human gingival fibroblasts plated on smooth and microgrooved/acid-etched titanium substrata. Material and methods. Three groups of titanium discs as NE0 (smooth Ti substrata), E15 (Ti substrata with microgrooves of $15{\mu}m$ of spacing and $3.5{\mu}m$ in depth and with further acidetching), and E30 (Ti substrata with microgrooves of $30{\mu}m$ spacing and $3.5{\mu}m$ in depth and with further acid-etching) served as the human gingival fibroblasts' substrata. Viability and proliferation of fibroblasts were determined using an XTT assay. Gene expressions of fibronectin, ${\alpha}5$ integrin, CDK4, and $p27^{kip}$ were analyzed in RT-PCR. Cell-substratum interactions were analyzed in SEM. Results. From the XTT assay at 24 h incubation, the mean optical density (OD) value of E15 was significantly greater than the values of E30 and NE0. At 48 and 96 h however, the mean OD values of E30 were significantly greater than the values of E15 and NE0. No differences in the expression of PCR transcripts at 96 h incubations were noted between groups, whereas at 48 h, an unexpected increase in the expression of all the transcripts were noted in E15 compared with other two groups. Fibroblasts were observed to orient and adhere inside the microgrooves. Conclusion. Micropatterned grooves and acid-etching on Ti substrata alter viability and gene expression of adhered human gingival fibroblasts.