• Title/Summary/Keyword: XTT

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Study on the Protective Effect of Nelumbo nucifera GAERTN Extract on Cultured Cerebral Neuroglial Cells Damaged by Hexavalent Chromium (연꽃추출물이 6가 크롬으로 유도된 세포독성에 대한 보호효과에 관한 연구)

  • Seo, Young-Mi;Park, Yun-Jum;Choi, Yu-Sun
    • FLOWER RESEARCH JOURNAL
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    • v.17 no.4
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    • pp.242-245
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    • 2009
  • In order to investigate the cytotoxic effect of hexavalent chromium ($Cr0_3$) and the protective effect of Nelumbo nucifera GAERTN (NNG) extract, cultured cerebral neuroglial cells (C6 glioma cells) were treated with $4{\sim}55{\mu}M$ concentrations of $Cr0_3$ for 48 hours. Cell viability was measured by XTT assay. The superoxide dismutase (SOD)-like activity for the antioxidant effect was also examined on the extract of NNG stamen. In this study, $Cr0_3$ significantly decreased cell viability dose-dependently. The cytotoxicities of $XTT_{90}$ and $XTT_{50}$ determined with $10{\mu}M$ and $55{\mu}M$ of $Cr0_3$, respectively, showed that the $Cr0_3$ had highly toxic effect on cultured C6 glioma cells by the cytotoxic criteria. In the protective effect of NNG extract, the cell viability was significantly increased by the treatment of NNG extract, and NNG extract increased SOD-like activity. From these results, it is suggested that $Cr0_3$ showed highly toxic effect on cultured C6 glioma cell s and NNG extract was very effective in the protection of $Cr0_3$-mediated cytotoxicity by antioxidative effect in these cultures.

Cytotoxic Effect of Hexavalent Chromium on C6 Glioma Cells

  • Suk Seung-Han
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.261-265
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    • 2006
  • Toxic effect of Hexavalent chromium $(CrO_3)$ on various cells and organs has been well recognized. However, the mechanism and degree of cytotoxicity of $CrO_3$ remain unclear. This study was performed to examine the cytotoxicity of $CrO_3$ on $C_6$ glioma cells by measuring cell viability. The XTT assay, one of the sensitive methods to determine the cell viability, was taken to examine the viability of glioma cells treated with $CrO_3$. In this study, not only decreased the number of glioma cells but morphologic changes of them were noted and cell viability decreased in a time and dose-dependent manner after treated with various concentrations of $CrO_3$ for 48hours. $IC_{90}\;and\;IC_{50}$ values in XTT assay were determined at $25{\mu}M\;and\;55{\mu}M$ $CrO_3$, respectively. These results suggest that Hexavalent chromium has a highly cytotoxic effect and has a time and dose-dependent cytotoxicity on $C_6$ glioma cells.

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Inhibitory Effects of Scutellaria barbata D. Don on the Cell Proliferation of HeLa cells (반지연(半枝蓮)이 HeLa Cell의 증식억제(增殖抑制)와 사멸(死滅)에 미치는 영향(影響))

  • Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub;Ha, Jee-Yeun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.4
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    • pp.47-60
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    • 2006
  • Purpose : This study was conducted to investigate the inhibitory effects of Scutellaria barbata D. D on on the cell proliferation of HeLa Cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Scutellaria barbata D. D on solution for 24, 48 and 72 hours for the direct inhibitory effects of Scutellaria barbata D. D on. Then we examined the effect of Scutellaria barbata D. D on solution on the cell proliferation inhibition by XTT assay. DNA fragmentation, MAP kinase activity and caspase activity by FACS analysis in HeLa cells. Results : We found that the proliferation of HeLa cells was significantly decreased in Scutellaria barbata D. D on solution containing groups comparing with a control group in a concentration-dependant manner. When HeLa cells were cultivated for 24 hours with 5% Scutellaria barbata D. D on solution containing group, the percentage of HeLa cells with activated caspase was the highest. Scutellaria barbata D. D on solution reduced the MAP kinase activity of HeLa cells comparing with the control group. By the XTT assay, the cell's activity was decreased in 5% and 10% Scutellaria barbata D. D on solution containing groups in 24 and 72 hours cultivation and 10% group in 48 hours. DNA fragmentation and caspase-3 activity of HeLa cells, however, were changed insignificantly. Conclusion : From this study we could suggest that Scutellaria barbata D. D on is available to the inhibition and apoptosis of human cervical carcinoma cell line, HeLa cells in vitro.

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Anti-HIV-1 Viral Activity of Glycyrrhizin (감초탕(甘草湯)의 Human Immunodeficiency Virus-1 억제능(抑制能)에 관(關)한 연구(硏究))

  • Lee, Tae-kyun;Moon, Jun-Joon
    • The Journal of Dong Guk Oriental Medicine
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    • v.1
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    • pp.209-236
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    • 1992
  • The microculture XTT antiviral assay method is used to quantitate HIV-1 induced cytopathic effects as modulated by test substances. This relatively simple assay facilitated the safe and rapid determination of in vitro antiviral activity of selected chemicals as well as direct cytotoxicity. This experiment also confirmed that this system measures infection and subsequent viral replication in target cells and XTT formazan formations correlated with the accumulation of extracellular virions, as measured by quantitative HIV-1 induced syncytium foramtion. The present results with Glycyrrhizin using this in vitro culture system demonstrated that effective dose, EC50(the concentration at which increases XTT formazan production in infected cultures to 50% of that in untreated, uninfected controls) was 250ml. As comparison, AZT was included in this experiment and demonstrated that EC50 AZT of was 0.05g/ml, approximately 5,000 times more potent than Glycyrrhizin based on EC50 ratio's alone. However, this potency is limited by severe cytotoxicity of AZT, while Glycyrrhizin is approximately 16 times less toxic(IC50 of Glycyrrhizin 800 and AZT 51 g/ml). While AZT's anti-HIV-1 viral activity is mediated by inhibition of reverse transcriptase of the virus, Glycyrrhizin faild to demonstrate any inhibitory activity against reverse transcriptase. Further study is necessary in order to understand the precise mechanisms of Glycyrrhizin action against HIV-1 viruses. Althouth Glycyrrhizin is less effective antiviral agent than AZT, much less toxicity of Glycyrrhizin is desirable in terms of chronic treatment. Combination treatment of AZT and Glycyrrhizin may be therapeutically beneficial. Clinical effectiveness of two drug combination therapy for AIDS patient is unknown at this time. However, this experimental investigation presents the scientific rational basis for such therapeutic approach.

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Effects of Jingansikpung-tang and Gamijingansikpung-tang Water Extract on the Cultured Spinal Sensory Neurons (진간식풍탕 및 가미진간식풍탕 추출물이 배양 척수감각신경세포에 미치는 영향)

  • Seo Young Suk;Yun Sang Hak;Yeom Seung Ryong;Lee Su kyung;Shin Byung Cheul;Kwon Young Dal;Song Yung Sun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.374-379
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    • 2003
  • To evaluate the mechanism of oxidative damage by Xanthine oxidase(XO) and hypoxanthine(HX)-induced oxygen radicals, XTT assay was carried out. Neurofilament EIA and PKC activity were measured to evaluate the protective effect of Jingansikpung-tang(JST) and Gamijingansikpung-tang(GJST) water extract on cultured spinal sensory neurons damaged by XO/HX, after the cultured mouse spinal sensory neurons were preincubated with various concentrations of JST and GJST water extract for 3 hours prior to exposure of XO/HX. The results were XO/HX decreased significantly, in proportion to concentration and exposed time, the survival rate of the cultured mouse sensory neurons on XTT assay. And in proportion to concentration and exposed time on cultured spinal sensory neurons, XO/HX showed the quantitative decrease of neurofilament by EIA, increase of PKC activity, but JST and GJST showed the neuroprotective effects against decrease of neurofilament and increase of PKC activity by XO/HX. From the above results, it is concluded that XO/HX have a neurotoxic effect on cultured spinal sensory neurons and the herbs water extract, such as JST and GJST prevent the toxicity of XO/HX effectively.

Inhibitory Effects of Dangguijakyaksan on Cell Proliferation in HeLa Cells (당귀작약산(當歸芍藥散)이 HeLa Cell의 증식억제(增殖抑制)와 사멸(死滅)에 미치는 영향(影響))

  • Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub;Park, Myung-Won
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.34-48
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    • 2006
  • Purpose : This study was conducted to investigate the inhibitory effects of Dangguijakyaksan on cell proliferation in HeLa cells. Methods : Human uterine cervical carcinoma HeLa cells were cultured in the 1%, 5% and 10% concentration of Dangguijakyaksan extract solution for 24 hours, 48 hours and 72 hours for the direct inhibitory effects of Dangguijakyaksan. Afterwards, we executed the analysis of the effect of Dangguijakyaksan extract solution on cell proliferation inhibition using XTT assay, molecular biological method through MAP kinase activity and FACS analysis of caspase activity in the HeLa cells. Results : After 24, 48 and 72 hours cultivation, Dangguijakyaksan extract solution group showed significant decrease of HeLa cells except 1% solution after 24 hours compared with the control group. In the FACS analysis, Dangguijakyaksan extract solution groups showed increase of caspase activity except 1% solution after 48 hours compared with the control group. In the XTT assay, the caspase-3 activities were increased in Dangguijakyaksan extract solution groups except 1% solution after 24 hours in a dose-dependent manner. In the XTT study, cell activities were significantly decreased in 10% Dangguijakyaksan extract solution groups after 48 and 72 hours cultivation compared with the control group. In all Dangguijakyaksan extract solution groups, The activities of MAP kinase were decreased after 24, 48 and 72 hours cultivation compared with the control group. Conclusion : It could be concluded that Dangguijakyaksan is available to the inhibition of proliferation of human cervical carcinoma cell line in vitro.

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TWO COLORIMETRIC ASSAYS VERIFY THAT CALCIUM SULFATE PROMOTES PROLIFERATING ACTIVITY OF HUMAN GINGIVAL FIBROBLASTS

  • Chae, Min;Kim, Su-Yeon;Kim, Soo-Yeon;Lee, Suk-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.3
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    • pp.382-388
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    • 2007
  • Statement of problem. The role of calcium sulfate in stimulating the growth of gingival soft tissue has been reported in few studies. Such a unique property of calcium sulfate could serve as a trouble-solving broker in compensating for the lack of soft tissues in various oral surgeries. Purpose. The purpose of this study was to compare the proliferating activities of human gingival fibroblasts seeded on various bone graft barrier materials of calcium sulfate, collagen, and polytetrafluorethylene (PTFE). Material and methods. Two calcium sulfates ($CAPSET^{(R)}$. and $CalForma^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA), a resorbable natural collagen ($Bio-Gide^{(R)}$, Geistlich Pharma Ag., Wolhusen, Switzerland), and a non-resorbable PTFE ($TefGen-FD^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA) served as the human gingival fibroblasts' substrates and comprised the four experimental groups, whereas the untreated floors of culture plastics were used in the control group, in this study. Cells were trypsinized, seeded, and incubated for 48 h. The proliferating activities of fibroblasts were determined by XTT and SRB assay and absorbance (optical density, OD) was measured. One-way ANOVA was used to analyze the differences in the mean OD values between the groups of CAPSET, CalForma, Bio-Gide, TefGen, and the control (p<0.05). Results. From the XTT assay, the mean OD value of the control group, the highest, was significantly greater than that of any of the four experimental groups followed by CalForma, CAPSET, TefGen, and Bio-Gide. Further, the mean OD value of CalForma, was significantly greater compared to that of Bio-Gide. From the SRB assay, Calforma showed the highest mean OD value, which was significantly greater than that of any other groups, followed by the control, CAPSET, Bio-Gide, and TefGen. The mean OD values of both the control and CAPSET were significantly greater compared to that of TefGen (p<0.05). Conclusion. Assessment of the viability and proliferation of cultured fibroblasts seeded and incubated for 48 h on various barrier-material substrates using XTT and SRB assay showed that calcium sulfate $CalForma^{(R)}$ promotes the proliferating activity of human gingival fibroblasts.

Biocompatibility of experimental mixture of mineral trioxide aggregate and glass ionomer cement (Glass ionomer cement와 혼합한 mineral trioxide aggregate의 생체친화성)

  • Oh, Min-Jae;Jeong, Yu-Na;Bae, In-Ho;Yang, So-Young;Park, Bum-Jun;Koh, Jeong-Tae;Hwang, Yun-Chan;Hwang, In-Nam;Oh, Won-Mann
    • Restorative Dentistry and Endodontics
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    • v.35 no.5
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    • pp.359-367
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    • 2010
  • Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.

COMPARISON OF BIOCOMPATIBILITY OF FOUR ROOT PERFORATION REPAIR MATERIALS (치근 천공 치료 재료의 생체친화성의 비교)

  • Kang, Min-Kyung;Bae, In-Ho;Koh, Jeong-Tae;Hwang, Yun-Chan;Hwang, In-Nam;Oh, Won-Mann
    • Restorative Dentistry and Endodontics
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    • v.34 no.3
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    • pp.192-198
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    • 2009
  • This study was carried out in order to determine in vitro biocompatibility of white mineral trioxide aggregate (MTA), and to compare it with that of the commonly used materials, i. e. calcium hydroxide liner (Dycal), glass ionomer cement (GIC), and Portland cement which has a similar composition of MTA. To assess the biocompatibility of each material, cytotoxicity was examined using MG-63 cells. The degree of cytotoxicity was evaluated by scanning electron microscopy (SEM) and a colorimetric method, based on reduction of the tetrazolium salt 2,3 bis {2methoxy 4nitro 5[(sulfenylamino) carbonyl] 2H tetrazolium hydroxide} (XTT) assay. The results of SEM revealed the cells in contact with GIC, MTA. and Portland cement at 1 and 3 days were apparently healthy. In contrast, cells in the presence of Dycal appeared rounded and detached. In XTT assay, the cellular activities of the cells incubated with all the test materials except Dycal were similar, which corresponded with the SEM observation. The present study supports the view that MTA is a very biocompatible root perforation repair material. It also suggests that cellular response of Portland cement and GIC are very similar to that of MTA.

Protective Effect of Kaempferol on Cultured Neuroglial Cells Damaged by Induction of Ischemia-like Condition

  • Son, Young-Woo;Choi, Yu-Ran;Seo, Young-Mi
    • Biomedical Science Letters
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    • v.23 no.4
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    • pp.339-347
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    • 2017
  • This study was performed to evaluate the cytotoxicity induced by ischemia-like condition (ILC) in cultured neuroglial cells (C6 glioma cells). The protective effect of kaempferol (KAE), flavonoid against the cytotoxicity induced by ILC induction was assessed. In addition, antioxidative effects of KAE were done by colorimetric assays. Cell viability and the antioxidative effects such as DPPH-radical scavenging activity, superoxide dismutase (SOD)-like activity and inhibitory activity of lipid peroxidation (LP) were analyzed. ILC induction decreased cell viability in a dose-dependent manner, and the $XTT_{90}$ value (low cytotoxicity value) and $XTT_{50}$ value (high cytotoxicity value) were determined during ILC induction for 15 and 40 minutes, respectively. The butylated hydroxytoluene (BHT) antioxidant significantly increased cell viability damaged by the ILC-induced cytotoxicity. In the protective effect of KAE on ILC-induced cytotoxicity, KAE protected the ILC-induced cytotoxicity by the significant increase of cell viability, and also it showed DPPH-radical scavenging ability, SOD-like ability and inhibitory ability of LP. From these results, it is suggested that ILC induction showed cytotoxicity in these cultures and the oxidative stress is involved in the ILC-induced cytotoxicity. While, KAE prevented ILC-induced cytotoxicity by antioxidative effects. In conclusion, natural products like KAE may be a putative therapeutic agent for the treatment of disease associated with oxidative stress such as ischemia.