Predictive performance of the Vitrigel-eye irritancy test method using 118 chemicals

J Appl Toxicol. 2016 Aug;36(8):1025-37. doi: 10.1002/jat.3254. Epub 2015 Oct 15.

Abstract

We recently developed a novel Vitrigel-eye irritancy test (EIT) method. The Vitrigel-EIT method is composed of two parts, i.e., the construction of a human corneal epithelium (HCE) model in a collagen vitrigel membrane chamber and the prediction of eye irritancy by analyzing the time-dependent profile of transepithelial electrical resistance values for 3 min after exposing a chemical to the HCE model. In this study, we estimated the predictive performance of Vitrigel-EIT method by testing a total of 118 chemicals. The category determined by the Vitrigel-EIT method in comparison to the globally harmonized system classification revealed that the sensitivity, specificity and accuracy were 90.1%, 65.9% and 80.5%, respectively. Here, five of seven false-negative chemicals were acidic chemicals inducing the irregular rising of transepithelial electrical resistance values. In case of eliminating the test chemical solutions showing pH 5 or lower, the sensitivity, specificity and accuracy were improved to 96.8%, 67.4% and 84.4%, respectively. Meanwhile, nine of 16 false-positive chemicals were classified irritant by the US Environmental Protection Agency. In addition, the disappearance of ZO-1, a tight junction-associated protein and MUC1, a cell membrane-spanning mucin was immunohistologically confirmed in the HCE models after exposing not only eye irritant chemicals but also false-positive chemicals, suggesting that such false-positive chemicals have an eye irritant potential. These data demonstrated that the Vitrigel-EIT method could provide excellent predictive performance to judge the widespread eye irritancy, including very mild irritant chemicals. We hope that the Vitrigel-EIT method contributes to the development of safe commodity chemicals. Copyright © 2015 The Authors. Journal of Applied Toxicology published by John Wiley & Sons Ltd.

Keywords: HCE T cells; collagen vitrigel membrane; corneal epithelium; eye irritation test; predictive performance; transepithelial electrical resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Testing Alternatives / methods*
  • Cells, Cultured
  • Collagen / chemistry
  • Electric Impedance
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Epithelium, Corneal / cytology
  • Epithelium, Corneal / drug effects
  • Epithelium, Corneal / metabolism
  • Eye / drug effects*
  • Eye / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Irritants / toxicity*
  • Models, Biological
  • Mucin-1 / genetics
  • Mucin-1 / metabolism
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tight Junctions / drug effects
  • Tight Junctions / metabolism
  • Toxicity Tests / methods*
  • United States
  • United States Environmental Protection Agency

Substances

  • Irritants
  • MUC1 protein, human
  • Mucin-1
  • Collagen