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.

Replacing the Draize eye test: Impedance spectroscopy as a 3R method to discriminate between all GHS categories for eye irritation

“…Although toluene and citrate are partly classified as no category substances, this test suggests a much stronger irritative capacity of both substances. This finding is also supported by several studies that show strong irritative effects of toluene and citrate 23 – 25 . In addition, toluene is categorized as a skin irritant by the European Chemcials Agency.…”

“…The European Union for example did not include the subcategories. TEER measurements at 12.5 Hz have shown to be a profound method to evaluate 118 chemicals for eye irritation 23 . In this study, however only three minutes after the application were measured not allowing for further evaluation of the reversibility of effects.…”

Collagen vitrigel promotes hepatocytic differentiation of induced pluripotent stem cells into functional hepatocyte-like cells

“…A collagen vitrigel (CV) membrane is composed of high-density collagen fibrils equivalent to connective tissues in vivo and possesses excellent transparency and permeability of protein with high molecular weight, which has been described to be utilized as a cell culture scaffolds (Takezawa et al, 2007; Wang and Takezawa, 2005). Collagen vitrigel membrane was reported to facilitate three dimensional formation of human corneal epithelial cells, and was useful to induce HepG2 cells to form hepatic structures that exhibited CYP3A4 activity and supported liver-specific functions of hepatocytes in long-term culture (Oshikata-Miyazaki and Takezawa, 2016; Watari et al, 2018; Yamaguchi et al, 2013, 2016).…”

A New in Vitro Model of GDLD by Knocking OutTACSTD2and Its Paralogous GeneEpCAMin Human Corneal Epithelial Cells

“…First, HCE-T cells have been utilized in numerous studies as a reliable in vitro model of corneal epithelial cells. 26,27 Second, HCE-T cells express major components of tight junction proteins, including ZO-1, occludin, and CLDN. 28 Third, HCE-T cells express the same set of CLDN proteins (CLDN1, 4, and 7) as in vivo corneal epithelial cells 29 ; this suggests that HCE-T cells still maintain their original cellular properties, at least with respect to epithelial barrier function.…”