title: Predictive performance of the Vitrigel-eye irritancy test method using 118 chemicals.
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.
authors.affiliation: Division of Animal Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.|Isehara Research Laboratory, Technology and Development Division, Kanto Chemical Co., Inc., Isehara, Kanagawa, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamaguchi
authors.affiliation: Japanese Center for the Validation of Alternative Methods (JaCVAM), Biological Safety Research Center, National Institute of Hearth Sciences, Setagaya, Tokyo, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Kojima
authors.affiliation: Division of Animal Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Takezawa
mesh_terms: D000826:Animal Testing Alternatives / Q000379:methods*; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D017097:Electric Impedance; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D019573:Epithelium, Corneal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005123:Eye / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007509:Irritants / Q000633:toxicity*; D008954:Models, Biological; D018396:Mucin-1 / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods*; D014481:United States; D014484:United States Environmental Protection Agency
chemical_list: D007509:Irritants; C060192:MUC1 protein, human; D018396:Mucin-1; D003094:Collagen
keywords: HCE T cells; collagen vitrigel membrane; corneal epithelium; eye irritation test; predictive performance; transepithelial electrical resistance