• Environmental Analysis Health and Toxicology
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• v.9 no.1_2
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• pp.19-23
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• 1994

The tetrazolium dye, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), is reduced by live but not dead cell, and this reaction is used as the end point in a rapid drug screening assay. It can also be used for accurate determinations of drug sensitivity but only if a quantative relationship is established between cell number and MTT-formazan production. Several conditions were examined to devise an in vitro assay method in primary cultured hepatocytes, such as optimum wavelength, optimal MTT concentration, optimal incubation time, and cell density.

• Restorative Dentistry and Endodontics
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• v.31 no.5
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• pp.398-408
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• 2006

The purpose of this study was to verify the usefulness of MTT analysis as a tool of measurement of the periodontal ligament cell viability from the extracted rat molar. A total of 80 Sprague-Dawley white female rat of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted under Ketamine anesthesia. Twenty-four teeth of each group (divided as five groups depending upon the time-lapse after extraction such as Immediate, 10, 20, 40 and 60 minutes) were immersed in $200{\mu}l$ of MTT solution (0.5 mg/ml) and processed for optical density measurements. Another 10 teeth of each group were treated as same as above and sectioned at $10{\mu}m$ for microscopic examination. All measurements values were divided by the value of hematoxylin-eosin staining which represented the volume of each corresponding samples. Immediate and 10 minute groups showed highest MTT values followed by 20, 40, and 60 minutes consecutively. Statistical significance (p<0.05) existed between all groups except in immediate versus 10 minute groups and 40 versus 60 minutes. Histological findings also showed similar findings with MTT results in crystal shape and crystal numbers between the experimental groups. These data indicate that in vivo MTT analysis nay be of value for evaluation of the periodontal ligament cell viability without time- consuming cell culturing processes.

• BMB Reports
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• v.38 no.3
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• pp.366-369
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• 2005

NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450 and also catalyzes the one-electron reduction of many drugs and foreign compounds. Various spectrophotometric assays have been performed to examine electron-accepting properties of CPR and its ability to reduce cytochrome $b_5$, cytochrome c, and ferricyanide. In this report, reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by CPR has been assessed as a method for monitoring CPR activity. The principle advantage of this substance is that the reduction of MTT can be assayed directly in the reaction medium by a continuous spectrophotometric method. The electrons released from NADPH by CPR were transferred to MTT. MTT reduction activity was then assessed spectrophotometrically by measuring the increase of $A_{610}$. MTT reduction followed classical Michaelis-Menten kinetics ($K_m\;=\;20\;{\mu}M$, $k_{cat}\;=\;1,910\;min^{-1}$). This method offers the advantages of a commercially available substrate and short analysis time by a simple measurement of enzymatic activity of CPR.

• Toxicological Research
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• v.9 no.1
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• pp.45-60
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• 1993

The present study was carried out to evaluate the cytotoxicity of cadmium on cultured rat fibroblasts. The colorimetric assays of neutral red and tetrazolium MTT, the lactatedehydrogenase activity, the amounts of total protein, the rate of DNA synthesis, the amounts of unscheduled DNA synthesis, the frequency of sister chromatid exchange, the releasing rate of intracellular calcium, and light and electron microscopic studies were performed on cultured rat fibroblasts maintained in the media containing various concentrations of cadmium. The results were as follows: The neutral red(NR) and MTT values were decreased dose-dependently by cadmium, and the NR90, NR50, MTT90 and MTT50 values of cadmium were 0.2mM, 21.5mM, 1.0Mm and 60.0Mm, respectively.

• Environmental Analysis Health and Toxicology
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• v.17 no.3
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• pp.253-259
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• 2002

The phototoxicity inhibitory activity of 15 natural products having antiinflammatory effect was screened by three in vitro methods: yeast growth inhibition test with Candida albicans, RBC photohemolysis and MTT assay. We induced phototoxic reaction by irradiating UVA (365 nm) on chlorpromazine (CPZ) that has been widely documented as phototoxic agent in clinical and experimental studies and then observed the effects of the natural products after treating them with CPZ. In yeast growth inhibition test, X. stramonium showed the inhibitory effect on the UVA phototoxicity and E. officinalis, Yeast, P. suffruticosa showed phototoxicity inhibitory effect in that their ％ hemolysis compared with control were 36.14${\pm}$ 2.69, 42.82${\pm}$1.35, 36.41${\pm}$0.48 on UVA. In MTT assay, all tested natural products increased cell viability compared with the control.

• Environmental Analysis Health and Toxicology
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• v.15 no.1_2
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• pp.13-18
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• 2000

A few in vitro methods were developed to compare the result on the phototoxicity of phenothiazines. By the MTT assay, the Candida test, and the RBC photohemolysis, the phototoxicities of UVA and UVB irradiation were measured. This paper presents the comparisons of methods which are effective to measure the phototoxicities of the chemicals causing phototoxicity and photoallergy. The tested chemicals of phenothiazines include Chlorpromazine, Promethazine, Perphenazine, Chlorprothixene, Trifluoperazine and Thioridazine. Each chemical represented variable results according to the test methods. MTT assay shows the most sensitive method.

• Restorative Dentistry and Endodontics
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• v.31 no.3
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• pp.192-202
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• 2006

The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.1
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• pp.187-199
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• 2009

Objectives : The electroacupuncture was generally has been used in oriental medicine doctors. In recent years, there have been a few studies about safety and stability of acupuncture needle in itself, but then again research of acupuncture needle safety and stability on applying electroacupuncture have been insignificant. Therefore we investigated the safety and change in mechanical characteristic of acupuncture needle in electroacupuncture therapy. Methods : We observed mechanical characteristic change by SEM(Scanning Electron Microscope) and EDX(Energy Dispersive X-ray Spectroscopy), evaluated the hardness by vickers hardness tester. We used MTT assay and cell stain to study about biocompatibility of electroacupuncture. Results : In this study, any corrosion of material, alternation of elements, and change of hardness were not observed in surface analysis using SEM and EDX. In cytotoxity evaluation using MTT assay and cell stain, cell survival rate was low when practicing the electroacupuncture for more than 3 hours. Conclusions : Change of mechanical property was not observed based on the test results using surface analysis and hardness estimation by the electroacupuncture. And considering the biocompatibility, electroacupuncture was thought to be safe in an hour based on cytotoxity evaluation using MTT assay and cell stain.

• Journal of Technologic Dentistry
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• v.31 no.3
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• pp.21-26
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• 2009

Purpose: Standards of alloy for porcelain fused to metal crown be classified by metallic factor and biological factor. Metallic factors consist of stability of alloy composition and mechanical strength and surface characteristics for chemical bond. Biological factors be considered properties of metallic elements and problems originated by toxicity and hypersensitive reaction. Alloys considered such controversial points are the most suitable alloy for dental instrument. Method: Alloys added Be and Nb using Ni-Cr alloy which has been widely used for dental instrument be selected and classified experimental group. Non-addition Be and Nb to Ni-Cr alloy classify control group and addition Be alloy is Be-experimental group, addition Nb alloy is Nb-experimental group. Specimens for cytotoxicity analysis gave effect to washing and sterilization. and then made an experiment on elution with cell medium after disinfection. It conducted specimens within cell medium with 24hours, 48hours, 72hours, respectively. It cultured human dermal fibroblast(HDF) using cell medium for cytotoxicity test and then investigated elution rate through spectroscopic analysis by MTT-assay. Result: As results of cytotoxicity test by MTT-assay, cultured cell rate of VII measured more low numerical value within elution medium for 24hours focused on control group. Also, cultured cell rate of K3 alloys observed low value for 48hours, 72hours than value of control group. Conclusion: According to final result that synthesize above results, Ni-Cr alloy added Be and Ni has little difference in Cytotoxicity by MTT-assay.

• The Journal of Korean Medicine
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• v.20 no.1 s.37
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• pp.91-105
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• 1999

The effects of Yinjin and Yinjinsaryongsangagambang on a DNA damaging agent, etoposide-induced apoptosis, cell viability, cell cycle progression, and mRNA expression of apoptosis-related genes of human hepatocyte cell line HepG2 were investigated using tryphan blue exclusion assay, MTT assay, flow cytometry, immunocytometric analysis of PCNA, and quantitative RT-PCR analysis. MTT assay showed that Yinjin and Yinjinsaryongsangagambang increases cellular viability of HepG2 cells in a dosage-dependent manner. Stimulation of cell cycle progression by Yinjin or Yinjinsaryongsangagambang was detected by flow cytometric analysis of the DNA content and immunocytometric analysis of PCNA expression. A significant reduction of a DNA-damaging agent, etoposide-induced apoptosis were found in both Yinjin and Yinjinsaryongsangagambang-treated cells in dosage-dependent manner. In overall, 3-fold reduction of apoptosis was recognized in $10.0\;{\mu}g/ml$ of Yinjin or Yinjinsaryongsangagambang-treated cells compared to untreated cells. Although the difference is not significant, Yinjinsaryongsangagambang showed slightly higher effect on the inhibition of apoptosis than Yinjin. From flow cytometric analysis of apoptosis, while 39.9% of untreated cells showed etoposide-induced apoptotic cell death, only 19.6% or 17.4% of Yinjin or Yinjinsaryongsangagambang-treated cells were fond at apoptotic sub G1 phase, respectively. Interestingly, strong induction of Gadd45-mRNA was observed from Yinjin or Yinjinsaryongsangagambang-treated cells. However, no changes in expression levels of p53 and Waf1 were detected, demonstrating that induction of Gadd45 mRNA expression by Yinjin or Yinjinsaryongsangagambang occurs by p53-independent mechanism. Marked mRNA inductions of two apoptosis-inhibiting genes, Bcl-2 and Bcl- XL, were found in both Yinjin or Yinjinsaryongsangagambang-treated HepG2 cells while no changes was detected in expression levels of an apoptosis-promoting gene, Bax.