The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay is a rapid, cheap, screening test for the in vitro anti-tuberculous activity of chalcones

2014 ◽  
Vol 104 ◽  
pp. 72-78 ◽  
Author(s):  
Suventha Moodley ◽  
Neil A. Koorbanally ◽  
Thrineshen Moodley ◽  
Deresh Ramjugernath ◽  
Manormoney Pillay
2018 ◽  
Vol 10 (04) ◽  
pp. 380-386 ◽  
Author(s):  
Thayanidhi Premamalini ◽  
Subramanian Anitha ◽  
Kanniappan Mohanapriya ◽  
Anupma Jyoti Kindo

ABSTRACT BACKGROUND: Invasive infections due to Trichosporon spp. have increased recently and are frequently associated with indwelling medical devices. Such infections which are associated with biofilm formation do not respond to the routinely used antifungal agents and are often persistent, associated with high mortality rate. Various methods have been described by researchers to evaluate and quantify the biofilm formation. AIM: This study was conducted to compare two methods of biofilm production by Trichosporon sp, i.e., test tube method with crystal violet (CV) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. MATERIALS AND METHODS: Seventy-two clinical isolates of Trichosporon spp. collected from various sources were considered for the study. The identity of all the isolates was genotypically confirmed by Trichosporon-specific polymerase chain reaction (PCR). The isolates were further speciated phenotypically using biochemical profile and growth characteristics which identified the isolates as Trichosporon asahii (64/72), Trichosporon asteroides (5/72), Trichosporon cutaneum (2/72), and Trichosporon mucoides (1/72). Biofilm production was then evaluated and compared by test tube-CV method and MTT assay. RESULTS: All the Trichosporon isolates produced biofilm by MTT assay, whereas only 42 (53.6%) of the isolates were detected to be biofilm producers by CV method. Furthermore, MTT assay could differentiate better between weak and moderate biofilm producers as compared to CV method. CONCLUSION: Hence, MTT assay is a reliable method for quantification of biofilm produced by Trichosporon spp. using 96-well microtiter plate.


2015 ◽  
Vol 754-755 ◽  
pp. 979-984
Author(s):  
Siti Mazatul Azwa Bt Saiyed Mohd Nurddin ◽  
Malek B. Selamat

The objective of the study was to determine the degree of biocompatibility of leucite glass-ceramics that have been produced from local high grade silica sand in terms of cytotoxicity and mutagenicity assays. In the present study, the cyctotoxicity and mutagenicity were studied using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay (MTT) and Ames Reverse Mutation. In the MTT assay, a dose response cytotoxicity of leucite sample was evaluated in L929 cells. The cells were treated with the concentrations of 6.25, 12.5, 25.0, 50.00, 100.00 and 200.00 mg/ml of the leucite sample for 24 hours. The cytotoxicity was determined by assessing the cell viability through the reduction of tetrazolium salts (MTT). The mutagenenicity of leucite sample was evaluated inS. typhiriumTA98. TA100, TA1535, TA1537 andE. coliWP2 in the Ames Reverse Mutation assay. Mutagenic effects were evaluated by comparing the mean number of revertant colonies of each extract concentraction with mean number of revertant colonies of the negative control. In results of MTT assay evaluated that the leucite did not show a cytotoxic effect at all concentrations under the condition of the study. Ames Reverse Mutation assay result proven that the leucite sample did not demonstrate a mutagenic effect under the condition of this study withSalmonella typhimuriumandEscherichia coli.


Author(s):  
Jos� Manuel Mart�nez-Martos ◽  
Mar�a Jes�s Ram�rez-Exp�sito ◽  
Mar�a Dolores Mayas-Torres ◽  
Mar�a Jes�s Garc�a-L�pez ◽  
Manuel Ram�rez-S�nchez

2020 ◽  
Vol 85 (3) ◽  
pp. 739-742
Author(s):  
Hideaki Shiraishi ◽  
Akiko Toyoda

ABSTRACT The applicability of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to an industrially valuable filamentous cyanobacterium Arthrospira platensis was examined. When it was applied to A. platensis NIES-39, as few as 10 viable trichomes were quantitatively detected. However, depending on the experimental conditions, it also generated artifactual viability signals. The results should help clarify the scope and limits of the MTT assay in viability analysis.


2021 ◽  
Vol 9 (3) ◽  
Author(s):  
Ludmil Benov

Reduction of the water-soluble tetrazolium salt 3-(4,5-dimethylthiazol)-2,5 diphenyl-tetrazolium bromide (MTT) to purple, water-insoluble formazan is commonly used for assessment of cell viability and proliferation. Spectrophotometric detection of formazan requires its solubilization.


2018 ◽  
Vol 13 (2) ◽  
pp. 101 ◽  
Author(s):  
Ernie Halimatushadyah ◽  
Muhammad Da’i ◽  
Muhammad Nursid

Teripang Holothuria atra merupakan biota laut yang banyak ditemukan di perairan Indonesia yang termasuk dalam filum Echinodermata dan berpotensi sebagai antikanker. Penelitian ini bertujuan untuk mengetahui sitotoksisitas dan induksi apoptosis ekstrak etanol teripang H. atra secara in vitro terhadap beberapa sel lestari. Pengujian sitotoksisitas dilakukan dengan metode MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) menggunakan sel HeLa, T47D, WiDr dan sel normal Vero, sedangkan uji induksi apoptosis dilakukan terhadap sel dengan hasil uji sitotoksisitas terbaik menggunakan metode flowcytometry dan double staining. Hasil penelitian menunjukkan bahwa ekstrak etanol  teripang H. atra mampu menghambat pertumbuhan sel kanker HeLa, T47D dan WiDr. Nilai IC50 ekstrak H. atra terhadap ketiga sel tersebut masing-masing sebesar 41,06±4,21; 20,89±1,55; 26,50±4,43 µg/ml tetapi esktrak tersebut memiliki sitotoksisitas  yang lebih rendah terhadap sel Vero (IC50 sebesar 128,00). Analisis flowcytometry dan double staining pada sel T47D memperlihatkan bahwa ekstrak etanol teripang H.atra mampu menginduksi apoptosis pada sel tersebut.


Author(s):  
Muhammad Arfat Yameen ◽  
Amir Zeb ◽  
Raza E Mustafa ◽  
Sana Mushtaq ◽  
Nargis Aman ◽  
...  

Background: Incoherent use of antibiotics has led toward resistance in MRSA, which is becoming multidrugresistant with high rate of virulence in the community and hospital settings. Objective: Synergistic anti-MRSA activity was investigated in this study for hybrid material composite spheres of amoxicillin, Ag nanoparticles and chitosan which were prepared by one-step synthesis method and various characterizations were performed. Methods: Antimicrobial-susceptibility assay on MRSA was achieved by disc diffusion and agar dilution techniques while agar well diffusion was used for hybrid composite spheres. The in vitro and cytotoxicity studies was done by skin abrasion mouse model and MTT assay on RD cell respectively. Results: All isolates were resistant with the tested antibiotics except vancomycin. MIC against MRSA showed high resistance with amoxicillin from 4 to 128 mg L-1. The mean diameter of chitosan spheres and Ag nanoparticles was 02 mm and 277 nm respectively. Morphology of spheres was uneven, varied, porous and irregular in SEM and Ag nanoparticles presence and formation was also seen in micrograph. No substantial interface among drug, nanoparticles and polymer was found in XRD and IR showed characteristic peaks of all compound in the formulation. The in vitro assay showed augmented anti-MRSA activity with amoxicillin loaded hybrid composite spheres (22-29 mm). A significant reduction in microbial burden (~6.5 log10 CFU ml-1) was seen in vivo with loaded hybrid composite spheres formulation. The MTT assay indicated no potential cytotoxicity with hybrid composite spheres. Conclusion: Synergistic effect, amoxicillin, new hybrid formulation, anti-MRSA activity, composite spheres. nanoparticles.


1991 ◽  
Vol 19 (3) ◽  
pp. 352-360
Author(s):  
Kazuhiko Otoguro ◽  
Kanki Komiyama ◽  
Satoshi Ωmura ◽  
Charles A. Tyson

Isolated hepatocytes from male Sprague-Dawley rats suspended in culture medium supplemented with either 0.2 or 2% bovine serum albumin (BSA) were allowed to attach to collagen coated 96-well dishes. Ten test chemicals from the MEIC list and salicylic acid were added individually to the dishes, and at the end of 24 and 48 hours, cytotoxicity was determined by measuring MTT (tetrazolium salt) reduction (mitochondrial integrity) and total cellular protein using Coomassie blue dye (reflecting cell number). Total cellular lactate dehydrogenase activity was also determined in some experiments, as an indicator of plasma membrane integrity. The relative toxicities of the test chemicals were quantified by the estimation of EC10, EC20 and EC50 values for each parameter. Except for one chemical, digoxin, in the MTT assay, cytotoxic potency increased with incubation time. The hepatocytes tended to be more sensitive to the chemicals in medium containing 0.2% BSA than in medium containing 2% BSA. Simple linear regression analyses of the log transformed data from the MTT assay versus log oral LD50 in rats for the test chemicals gave the best results using EC10 at 24 hours (r2 = 0.86). With protein as the cytotoxic indicator, the best results were obtained with EC values in the medium containing 2% BSA, again at 24 hours (r2 = 0.83). These results suggest that the MTT and Coomassie blue dye assays could be useful indicators for testing the cytotoxic potential of chemicals in rat hepatocyte cultures.


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