Development of Respiratory Mucosal Irritation and Toxicity Screening Methods: Comparison of MTT and Colorimetric Resazurin-Based Assay in Juniper Publishers

JUNIPER PUBLISHERS-OPEN ACCESS JOURNAL OF TOXICOLOGY



Development of Respiratory Mucosal Irritation and Toxicity Screening Methods: Comparison of MTT and Colorimetric Resazurin-Based Assay


Authored by Remigius Uchenna Agu


Abstract
Availability of reliable and reproducible assay methods that use simple tools will facilitate research especially in resource-poor settings. The purpose of this study is to evaluate colorimetric resazurin reduction assay (CRA) as a possible tool in respiratory mucosal cell irritation and toxicity screening. It compared CRA with another well-accepted colorimetric cell viability assay method (3- [4, 5-Dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolinium bromide {MTT} assay). Cell viability results of treated cells were obtained using both assay methods and used to determine Z' factor, signal-to-background (S/B) ratio and IC50 values. The results show all the Z ' values obtained were positive except values from CRA for cells treated with triton-X 100 incubated for 30min and 2h. All the Z ' values for MTT shows the method is optimized (0.5< Z'<1.0) while most values for CRA were outside the acceptable range (Z'<0, Z'>1). MTT demonstrated large S/B ratio at both 30min and 2h for all the compounds. The range of values obtained for the S/B ratio is 12-57 for MTT and 1-3 for CRA. Furthermore, CRA failed to estimate IC50 for some of the compounds (BKC, 2h; SDS, 2h; BKC, 30min). The values obtained for Z' factor, S/B ratio and IC50 for both assays were at variance. This result demonstrates that CRA performed below acceptable assay standard and therefore may not be an appropriate method for human respiratory cell viability studies. Consequently, we conclude that CRA is not useful in human respiratory mucosal irritation and toxicity studies.

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