Development of a Convenient and Quantitative Method for Evaluating Photosensitizing Activity Using Thiazolyl Blue Formazan Dye

Overview

Journal Molecules

Publisher MDPI

Specialty Biology

Date 2024 Jun 19

PMID 38893346

Authors

Smee Kang

Yeong Ji Oh

Mi-Ri Kim

Yu Na Jung

Eiseul Song

Hyowon Lee

Jungil Hong

Affiliations

Soon will be listed here.

Abstract

Photosensitizers cause oxidative damages in various biological systems under light. In this study, the method for analyzing photosensitizing activity of various dietary and medicinal sources was developed using 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (thiazolyl blue formazan; MTT-F) as a probe. Significant and quantitative decolorization of MTT-F was observed in the presence of photosensitizers used in this study under light but not under dark conditions. The decolorization of MTT-F occurred irradiation time-, light intensity-, and photosensitizer concentration-dependently. The decolorized MTT-F was reversibly reduced by living cells; the LC-MS/MS results indicated the formation of oxidized products with -1 / of base peak from MTT-F, suggesting that MTT-F decolorized by photosensitizers was its corresponding tetrazolium. The present results indicate that MTT-F is a reliable probe for the quantitative analysis of photosensitizing activities, and the MTT-F-based method can be an useful tool for screening and evaluating photosensitizing properties of various compounds used in many industrial purposes.

References

Scotter M, Castle L . Chemical interactions between additives in foodstuffs: a review. Food Addit Contam. 2004; 21(2):93-124. DOI: 10.1080/02652030310001636912. View

Minnock A, Vernon D, Schofield J, Griffiths J, Parish J, Brown S . Photoinactivation of bacteria. Use of a cationic water-soluble zinc phthalocyanine to photoinactivate both gram-negative and gram-positive bacteria. J Photochem Photobiol B. 1996; 32(3):159-64. DOI: 10.1016/1011-1344(95)07148-2. View

Twentyman P, LUSCOMBE M . A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Br J Cancer. 1987; 56(3):279-85. PMC: 2002206. DOI: 10.1038/bjc.1987.190. View

Alloy M, Finch B, Ward C, Redman A, Bejarano A, Barron M . Recommendations for advancing test protocols examining the photo-induced toxicity of petroleum and polycyclic aromatic compounds. Aquat Toxicol. 2023; 256:106390. PMC: 10519366. DOI: 10.1016/j.aquatox.2022.106390. View

Rosenkranz A, Schmaldienst S, Stuhlmeier K, Chen W, Knapp W, Zlabinger G . A microplate assay for the detection of oxidative products using 2',7'-dichlorofluorescin-diacetate. J Immunol Methods. 1992; 156(1):39-45. DOI: 10.1016/0022-1759(92)90008-h. View

LABBE R, Vreman H, Stevenson D . Zinc protoporphyrin: A metabolite with a mission. Clin Chem. 1999; 45(12):2060-72. View

Sladowski D, Steer S, Clothier R, Balls M . An improved MTT assay. J Immunol Methods. 1993; 157(1-2):203-7. DOI: 10.1016/0022-1759(93)90088-o. View

Morgan D . Tetrazolium (MTT) assay for cellular viability and activity. Methods Mol Biol. 1998; 79:179-83. DOI: 10.1385/0-89603-448-8:179. View

Pushpan S, Venkatraman S, Anand V, Sankar J, Parmeswaran D, Ganesan S . Porphyrins in photodynamic therapy - a search for ideal photosensitizers. Curr Med Chem Anticancer Agents. 2003; 2(2):187-207. DOI: 10.2174/1568011023354137. View

10.

Wang Y, Wang Q, Zhan X, Wang F, Safdar M, He J . Visible light driven type II heterostructures and their enhanced photocatalysis properties: a review. Nanoscale. 2013; 5(18):8326-39. DOI: 10.1039/c3nr01577g. View

11.

Cardoso D, Libardi S, Skibsted L . Riboflavin as a photosensitizer. Effects on human health and food quality. Food Funct. 2012; 3(5):487-502. DOI: 10.1039/c2fo10246c. View

12.

Jin Y, Zhang X, Zhang B, Kang H, Du L, Li M . Nanostructures of an amphiphilic zinc phthalocyanine polymer conjugate for photodynamic therapy of psoriasis. Colloids Surf B Biointerfaces. 2015; 128:405-409. DOI: 10.1016/j.colsurfb.2015.02.038. View

13.

Lee J, Kim J, Ye Y, Kim T . Materials and device design for advanced phototherapy systems. Adv Drug Deliv Rev. 2022; 186:114339. DOI: 10.1016/j.addr.2022.114339. View

14.

Holder A, Goth-Goldstein R, Lucas D, Koshland C . Particle-induced artifacts in the MTT and LDH viability assays. Chem Res Toxicol. 2012; 25(9):1885-92. PMC: 3446248. DOI: 10.1021/tx3001708. View

15.

Buchczyk D, Klotz L, Lang K, Fritsch C, Sies H . High efficiency of 5-aminolevulinate-photodynamic treatment using UVA irradiation. Carcinogenesis. 2001; 22(6):879-83. DOI: 10.1093/carcin/22.6.879. View

16.

Konings E . Committee on food nutrition. Water-soluble vitamins. J AOAC Int. 2003; 86(1):147-9. View

17.

Ochsner M . Photophysical and photobiological processes in the photodynamic therapy of tumours. J Photochem Photobiol B. 1997; 39(1):1-18. DOI: 10.1016/s1011-1344(96)07428-3. View

18.

Luis C, Castano-Guerrero Y, Soares R, Sales G, Fernandes R . Avoiding the Interference of Doxorubicin with MTT Measurements on the MCF-7 Breast Cancer Cell Line. Methods Protoc. 2019; 2(2). PMC: 6632110. DOI: 10.3390/mps2020029. View

19.

Petty R, Sutherland L, Hunter E, Cree I . Comparison of MTT and ATP-based assays for the measurement of viable cell number. J Biolumin Chemilumin. 1995; 10(1):29-34. DOI: 10.1002/bio.1170100105. View

20.

Landi M . Commentary to: "Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds" by Hodges et al., Planta (1999) 207:604-611. Planta. 2017; 245(6):1067. DOI: 10.1007/s00425-017-2699-3. View