scholarly journals Improved Formazan Dissolution for Bacterial MTT Assay

2021 ◽  
Vol 9 (3) ◽  
Author(s):  
Ludmil Benov
Keyword(s):  
Cell Viability ◽  
Mtt Assay ◽  
Water Soluble ◽  
Tetrazolium Salt ◽  
Spectrophotometric Detection ◽  
Tetrazolium Bromide ◽  
Diphenyl Tetrazolium Bromide

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.

scholarly journals Comparative study of hyperpure chlorine dioxide with two other irrigants regarding the viability of periodontal ligament stem cells

2020 ◽  
Author(s):  
Orsolya Láng ◽  
Krisztina S. Nagy ◽  
Julia Láng ◽  
Katalin Perczel-Kovách ◽  
Anna Herczegh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Viability ◽  
Chlorine Dioxide ◽  
Periodontal Ligament ◽  
Water Soluble ◽  
Tetrazolium Salt ◽  
Stem Cell Markers ◽  
Periodontal Ligament Stem Cells ◽  
Cell Markers

Abstract Objectives Periodontal ligament stem cells (PDLSCs) have an underlined significance as their high proliferative capacity and multipotent differentiation provide an important therapeutic potential. The integrity of these cells is frequently disturbed by the routinely used irrigative compounds applied as periodontal or endodontic disinfectants (e.g., hydrogen peroxide (H2O2) and chlorhexidine (CHX)). Our objectives were (i) to monitor the cytotoxic effect of a novel dental irrigative compound, chlorine dioxide (ClO2), compared to two traditional agents (H2O2, CHX) on PDLSCs and (ii) to test whether the aging factor of PDLSC cultures determines cellular responsiveness to the chemicals tested. Methods Impedimetry (concentration-response study), WST-1 assays (WST = water soluble tetrazolium salt), and morphology analysis were performed to measure changes in cell viability induced by the 3 disinfectants; immunocytochemistry of stem cell markers (STRO-1, CD90, and CD105) measured the induced mesenchymal characteristics. Results Cell viability experiments demonstrated that the application of ClO2 does not lead to a significant decrease in viability of PLDSCs in concentrations used to kill microbes. On the contrary, traditional irrigants, H2O2, and CHX are highly toxic on PDLSCs. Aging of PLDSC cultures (passages 3 vs. 7) has characteristic effects on their responsiveness to these agents as the increased expression of mesenchymal stem cell markers turns to decreased. Conclusions and clinical relevance While the active ingredients of mouthwash (H2O2, CHX) applied in endodontic or periodontitis management have a serious toxic effect on PDLSCs, the novel hyperpure ClO2 is less toxic providing an environment favoring dental structure regenerations during disinfectant interventions.

A water-soluble tetrazolium salt useful for colorimetric cell viability assay

1999 ◽  
Vol 36 (2) ◽  
pp. 47-50 ◽  
Author(s):  
Hideyuki Tominaga ◽  
Munetaka Ishiyama ◽  
Fumio Ohseto ◽  
Kazumi Sasamoto ◽  
Tomoyuki Hamamoto ◽  
...  
Keyword(s):  
Cell Viability ◽  
Water Soluble ◽  
Tetrazolium Salt ◽  
Cell Viability Assay ◽  
Viability Assay

scholarly journals Interference of Antioxidant Flavonoids with MTT Tetrazolium Assay in a Cell Free System

2021 ◽  
pp. 76-83
Author(s):  
Ashwini Somayaji ◽  
C. S. Shastry
Keyword(s):  
Cell Viability ◽  
Mtt Assay ◽  
Mammalian Cells ◽  
Suitable Method ◽  
Radical Scavenger ◽  
Tetrazolium Salt ◽  
Srb Assay ◽  
Free System ◽  
Cell Free System ◽  
A Cell

Introduction: The tetrazolium salt 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) used extensively to measure the quantitative survival and proliferation of mammalian cells. The analysis is based upon the reduction of MTT by metabolically active cells to insoluble formazan crystals. Flavonoids are a large group of natural compounds found in plants with variable phenolic structures.  Flavonoids, as they are potential reducing agents, they act as a free radical scavenger. The aim of the study is to assess the reducing effect of some of the flavonoids on tetrazolium salt and their interference with the colorimetric analysis of MTT. The cell viability obtained from the MTT assay was compared with that of SRB assay in the determination of flavonoids cytotoxicity.  Materials and Methods: The present study examined the effect of few bio-flavonoids like Quercetin, EGCG, Rutin and Resveratrol to reduce MTT in the absence of cells under different experimental conditions such as concentration of flavonoids, incubation time and results were compared with SRB assay findings. The study also involves the analysis of flavonoid cytotoxicity on lung cancer cells NCIH-460 and NCIH-522 by MTT and SRB assay to establish the suitable cell viability assay for  flavonoids. Results: All the flavonoids showed the instant formation of the dark blue formazan salt in the absence of the cells with MTT assay. Whereas SRB assay of flavonoids in the absence of cells, results showed the absorbance similar to that of the blank, indicating that SRB did not interfere with flavonoids in a cell-free system. Conclusion: From the results, it is evident that MTT is not a suitable method to determine the effect of flavonoids on cell viability and proliferation as flavonoids itself reduces the MTT to formazan crystals. Study also suggests that SRB assay is more suitable method to determine the effect of flavonoids on cell viability.

Utility of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to measure mitochondrial activity in K+- and ATP-stimulated rodent cortex synaptosomes

2000 ◽  
Vol 27 (2) ◽  
pp. 103-107 ◽  
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
Keyword(s):  
Mtt Assay ◽  
Mitochondrial Activity ◽  
Tetrazolium Bromide ◽  
Diphenyl Tetrazolium Bromide

scholarly journals The use of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide–based colorimetric assay in the viability analysis of the filamentous cyanobacterium Arthrospira platensis

2020 ◽  
Vol 85 (3) ◽  
pp. 739-742
Author(s):  
Hideaki Shiraishi ◽  
Akiko Toyoda
Keyword(s):  
Mtt Assay ◽  
Colorimetric Assay ◽  
Arthrospira Platensis ◽  
Filamentous Cyanobacterium ◽  
Experimental Conditions ◽  
Viability Analysis ◽  
Tetrazolium Bromide ◽  
Diphenyl Tetrazolium Bromide

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.

scholarly journals Evaluation of 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide method for assessing biofilm formation in vitro by Trichosporon spp.

2018 ◽  
Vol 10 (04) ◽  
pp. 380-386 ◽  
Author(s):  
Thayanidhi Premamalini ◽  
Subramanian Anitha ◽  
Kanniappan Mohanapriya ◽  
Anupma Jyoti Kindo
Keyword(s):  
Mtt Assay ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Test Tube ◽  
Tetrazolium Bromide ◽  
Biofilm Production ◽  
Invasive Infections ◽  
Polymerase Chain ◽  
Diphenyl Tetrazolium Bromide

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.

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