Limitation of the MTT and XTT assays for measuring cell viability due to superoxide formation induced by nano-scale TiO 2

Toxicology in Vitro

Volumn 25, Issue 8, 2011, Pages 2147-2151

  Wang, Shuguang ^a   Yu, Hongtao ^b   Wickliffe, Jeffrey K ^a  

^a TULANE UNIVERSITY   (United States)

^b JACKSON STATE UNIVERSITY   (United States)

Author keywords

Cell viability; MTT XTT; Nano TiO 2; Superoxide; Tetrazolium

Indexed keywords

2,3 BIS(2 METHOXY 4 NITRO 2H TETRAZOLIUM BROMIDE); 3 (4,5 DIMETHYL 2 THIAZOLYL) 2,5 DIPHENYLTETRAZOLIUM BROMIDE; DYE; SUPEROXIDE; TETRAZOLIUM; TITANIUM DIOXIDE; TRYPTAN BLUE; UNCLASSIFIED DRUG;

ANIMAL CELL; ARTICLE; CELL CULTURE; CELL PROLIFERATION; CELL VIABILITY; COMPARATIVE EFFECTIVENESS; CONTROLLED STUDY; CYTOLOGY; CYTOTOXICITY; FEMALE; INTERMETHOD COMPARISON; NANOANALYSIS; NONHUMAN; PREDICTION;

ANIMALS; CELL SURVIVAL; CHO CELLS; COLORING AGENTS; CRICETINAE; CRICETULUS; NANOPARTICLES; SUPEROXIDES; TETRAZOLIUM SALTS; THIAZOLES; TITANIUM; TOXICITY TESTS;

CRICETULUS GRISEUS; MAMMALIA;

EID: 82655172470     PISSN: 08872333     EISSN: 18793177     Source Type: Journal    
DOI: 10.1016/j.tiv.2011.07.007     Document Type: Article

References (37)

1. Abe K., Matsuki K. Measurement of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction activity and lactate dehydrogenase release using MTT. Neurosci. Res. 2000, 38:325-329.
2. Altman S.A., Randers L., Rao G. Comparison of trypan blue dye exclusion and fluorometric assays for mammalian cell viability determinations. Biotechnol. Progress. 1993, 9(6):671-674.
3. Belyanskaya L., Manser P., Spohn P., Bruinink A., Wick P. The reliability and limits of the MTT reduction assay for carbon nanotubes-cell interaction. Carbon 2007, 45(13):2643-2648.
4. Berridge M.V., Herst P.M., Tan A.S. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol. Annu. Rev. 2005, 11:127-152.
5. Bielski B.H.J., Shiue G.G., Bajuk S. Reduction of nitro blue tetrazolium by CO2- and O2- radicals. J. Phys. Chem. 1980, 84:830-833.
6. Ganapathy-Kanniappan S., Geschwind J.F.H., Kunjithapatham R., Buijs M., Syed L.H., Rao P.P., Ota S., Vali M. The pyruvic acid analog 3-bromopyruvate interferes with the tetrazolium reagent MTS in the evaluation of cytotoxicity. Assay Drug Dev. Tech. 2010, 8(2):258-262.
7. Gormley A.J., Ghandehari H. Evaluation of Toxicity of Nanostructures in Biological Systems. Nanotoxicity Chichester 2009, 115-159. John Wiley & Sons Ltd, UK. S.C.C. Sahu, A. Daniel (Eds.).
8. Gurr J.R., Wang A.S., Chen C.H., Jan K.Y. Ultrafine titanium dioxide particles in the absence of photoactivation can induce oxidative damage to human bronchial epithelial cells. Toxicol. 2005, 213:66-73.
9. Han M., Li J.F., Tan Q., Sun Y.Y., Wang Y.Y. Limitations of the use of MTT assay for screening in drug discovery. J. Chin. Pharm. Sci. 2010, 19(3):195-200.
10. Hasen J., Bross P. A cellular viability assay to monitor drug toxicity. Meth. Mol. Biol. 2010, 648:303-311.
11. Heinrich P., Loffler G., Petrides P.E. Biochemie und Pathobiochemie (Springer-Lehrbuch) 2006, Springer, Berlin.
12. 2 nanoparticles in anatase and rutile crystal phase. Biol. Rrace Elem. Res. 2010, 10.1007/s12011-010-8707-0.
13. Johno H., Takahashi S., Kitamura M. Influences of acidic conditions on formatzan assay: a cautionary note. Appl. Biochem. Biotechnol. 2010, 162(6):1529-1535.
14. Kang S.-G., Lee D.-Y., Kang M.-L., Yoo H.-S. Biological characteristics of Chinese hamster ovary cells transfected with bovine Prnp. J. Vet. Sci. 2007, 8:131-137.
15. Kato T., Loo B.H., Yokomaku M., Butsugan Y., Sim K.Y., Fujishima A. Photoelectrochemical reduction of tetrazolium salts to formazans on surfaces of semiconductor powders in alcohol solutions. Spectr. Lett. 1995, 28(6):849-859.
16. Kroll A., Pillukat M.H., Hahn D., Schnekenburger J. Current in vitro methods in nanoparticle risk assessment: limitations and challenges. Eur. J. Pharm. Biopharm. 2009, 72(2):370-377.
17. Lipovsky A., Nitzan Y., Gedanken A., Lubart R. Antifungal activity of ZnO nanoparticles-the role of ROS mediated cell injury. Nanotechnology 2011, 22(10):105101.
18. Long T.C., Saleh N., Tilton R.D., Lowry G.V., Veronesi B. Titanium dioxide (P25) produces reactive oxygen species in immortalized brain microglia (BV2): implications for nanoparticle neurotoxicity. Environ. Sci. Technol. 2006, 40:4346-4352.
19. Long T.C., Tajuba J., Sama P., Saleh N., Swartz C., Parker J., Hester S., Lowry G.V., Veronesi B. Nanosize titanium dioxide stimulates reactive oxygen species in brain microglia and damages neurons in vitro. Environ. Health Perspect. 2007, 115:1631-1637.
20. Madesh M., Balasubramanian K.A. A microtiter plate assay for superoxide using MTT reduction method. Indian J. Biochem. Biophys. 1997, 34:535-539.
21. Madesh M., Balasubramanian K.A. Microtiter plate assay for superoxide dismutase using MTT reduction by superoxide. Indian J. Biochem. Biophys 1998, 35:184-188.
22. Marques E.P., Zhang J., Tse Y.H., Metcalfe R.A., Pietro W.J., Lever A.B.P. Surface electrochemistry of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTTBr) absorbed on a graphite electrode. J. Electroanal. Chem. 1995, 395:133-142.
23. Oritani T., Fukuhara N., Okajima T., Kitamura F., Ohsaka T. Electrochemical and spectroscopic studies on electron-transfer reaction between novel water-soluble tetrazolium salts and a superoxide ion. Inorg. Chim. Acta. 2004, 357:436-442.
24. Park M.V.D.Z., Lankveld D.P.K., van Loveren H., de Jong W.H. The status of in vitro toxicity studies in the risk assessment of nanomaterials. Nanomedicine 2009, 4(6):669-685.
25. Plumb J.A. Cell Sensitivity Assay: The MTT Assay. Cancer Cell Culture: Methods and Protocols 1999, 163-169. Humana Press, Inc, Totwa, NJ. S.P. Langdon (Ed.).
26. Qu Q.L., Zhang Y.G. Cytotoxic effects of activated carbon nanoparticles, silicon dioxide nanoparticles and titanium dioxide nanoparticles on human gastric carcinoma cell line BGC-823. Chin. J. Pharmacol. Toxicol. 2010, 24(6):481-487.
27. Roehm N.W., Rodgers G.H., Hatfield S.M., Glasebrook A.L. An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J. Immunol. Methods 1991, 142:257-265.
28. Shimamura T., Takamori A., Ukeda H., Nagata S., Sawamura M. Relationship between the reduction of tetrazolium salt XTT and DNA strand breakage with aminosugars. J. Agric. Food Chem. 2000, 48:1204-1209.
29. Shukla R.K., Sharma V., Pandey A.K., Singh S., Sultana S., Dhawan A. ROS-mediated genotoxicity induced by titanium dioxide nanoparticles in human epidermal cells. Toxicol. in vitro 2011, 25(1):231-241.
30. Singh S., Shi T., Duffin R., Albrecht C., van Berlo D., Höhr D., Fubini B., Martra G., Fenoglio I., Borm P.J.A., Schins R.P.F. Endocytosis, oxidative stress and IL-8 expression in human lung epithelial cells upon treatment with fine and ultrafine TiO2: Role of the specific surface area and of surface methylation of the particles. Toxicol. Appl. Pharmacol. 2007, 222(2):141-151.
31. Stewart R.S., Harris D.A. Mutational analysis of topological determinants in prion protein (PrP) and measurement of transmembrane and cytosolic PrP during prion infection. J. Biol. Chem. 2003, 278:45960-45968.
32. Sutherland M.W., Learmonth B.A. The tetrazolium dyes MTS and XTT provide new quantitative assays for superoxide and superoxide dismutase. Free Radic. Res. 1997, 27:283-289.
33. Tsukatani T., Higuchi T., Suenaga H., Akao T., Ishiyama M., Ezoe T., Matsumoto K. Colorimetric microbial viability assay based on reduction of water-soluble tetrazolium salts for antimicrobial susceptibility testing and screening of antimicrobial substances. Anal. Biochem. 2009, 393(1):117-125.
34. Voinov M.A., Pagán J.O.S., Morrison E., Smirnova T.I., Smirnov A.I. Surface-mediated production of hydroxyl radicals as a mechanism of iron oxide nanoparticle biotoxicity. J. Am. Chem. Soc. 2011, 133(1):35-41.
35. Wang H., Cheng H., Wang F., Wei D., Wang X. An improved 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay for evaluating the viability of Escherichia coli cells. J. Microbiol. Meth. 2010, 82(3):330-333.
36. Xue C., Liu W., Wu J., Yang X., Xu H. Chemoprotective effect of N-acetylcysteine (NAC) on cellular oxidative damages and apoptosis induced by nano titanium dioxide under UVA irradiation. Toxicol. in Vitro 2010, 25:110-116.
37. Zhu X., Chen J., Scheideler L., Reichl R., Geis-Gerstorfer J. Effects of topography and composition of titanium surface oxides on osteoblast responses. Biomaterials 2004, 25:4087-4103.