A human hemi-cornea model for eye irritation testing: quality control of production, reliability and predictive capacity

TY - JOUR

T1 - A human hemi-cornea model for eye irritation testing: quality control of production, reliability and predictive capacity

AU - Engelke, M

AU - Zorn-Kruppa, M

AU - Gabel, D

AU - Reisinger, K

AU - Rusche, B

AU - Mewes, K R

N1 - Copyright © 2012 Elsevier Ltd. All rights reserved.

PY - 2013/2/1

Y1 - 2013/2/1

N2 - We have developed a 3-dimensional human hemi-cornea which comprises an immortalized epithelial cell line and keratocytes embedded in a collagen stroma. In the present study, we have used MTT reduction of the whole tissue to clarify whether the production of this complex 3-D-model is transferable into other laboratories and whether these tissues can be constructed reproducibly. Our results demonstrate the reproducible production of the hemi-cornea model according to standard operation procedures using 15 independent batches of reconstructed hemi-cornea models in two independent laboratories each. Furthermore, the hemi-cornea tissues have been treated with 20 chemicals of different eye-irritating potential under blind conditions to assess the performance and limitations of our test system comparing three different prediction models. The most suitable prediction model revealed an overall in vitro-in vivo concordance of 80% and 70% in the participating laboratories, respectively, and an inter-laboratory concordance of 80%. Sensitivity of the test was 77% and specificity was between 57% and 86% to discriminate classified from non-classified chemicals. We conclude that additional physiologically relevant endpoints in both epithelium and stroma have to be developed for the reliable prediction of all GHS classes of eye irritation in one stand alone test system.

AB - We have developed a 3-dimensional human hemi-cornea which comprises an immortalized epithelial cell line and keratocytes embedded in a collagen stroma. In the present study, we have used MTT reduction of the whole tissue to clarify whether the production of this complex 3-D-model is transferable into other laboratories and whether these tissues can be constructed reproducibly. Our results demonstrate the reproducible production of the hemi-cornea model according to standard operation procedures using 15 independent batches of reconstructed hemi-cornea models in two independent laboratories each. Furthermore, the hemi-cornea tissues have been treated with 20 chemicals of different eye-irritating potential under blind conditions to assess the performance and limitations of our test system comparing three different prediction models. The most suitable prediction model revealed an overall in vitro-in vivo concordance of 80% and 70% in the participating laboratories, respectively, and an inter-laboratory concordance of 80%. Sensitivity of the test was 77% and specificity was between 57% and 86% to discriminate classified from non-classified chemicals. We conclude that additional physiologically relevant endpoints in both epithelium and stroma have to be developed for the reliable prediction of all GHS classes of eye irritation in one stand alone test system.

KW - Animal Testing Alternatives

KW - Cell Line

KW - Cell Survival

KW - Cornea

KW - Humans

KW - Irritants

KW - Models, Biological

KW - Quality Control

KW - Reproducibility of Results

U2 - 10.1016/j.tiv.2012.07.011

DO - 10.1016/j.tiv.2012.07.011

M3 - SCORING: Journal article

C2 - 22902307

VL - 27

SP - 458

EP - 468

JO - TOXICOL IN VITRO

JF - TOXICOL IN VITRO

SN - 0887-2333

IS - 1

ER -