Prevalidation trial for a novel in vitro eye irritation test using the reconstructed human cornea-like epithelial model, MCTT HCE

• Published in: Toxicology in vitro Vol. 39; pp. 58 - 67
• Main Authors: Yang, Hyeri, Kim, Da-eun, Jang, Won-Hee, An, Susun, Cho, Sun-A, Jung, Mi-Sook, Lee, Ji Eun, Yeo, Kyung-Wook, Koh, Sang Bum, Jeong, Tae-Cheon, Kang, Mi-Jeong, Chun, Young-Jin, Lee, Su-Hyon, Lim, Kyung-Min, Bae, SeungJin
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.03.2017; Elsevier Science Ltd
• Subjects: Animal Testing Alternatives; Cell Survival - drug effects; Cornea; Epithelium; Epithelium, Corneal - drug effects; Eye irritation test; Eyes & eyesight; Humans; In vitro methods and tests; In Vitro Techniques; Irritants - toxicity; Irritation; MCTT HCE; Predictive capacity; Reproducibility; Reproducibility of Results; Sensitivity; Sensitivity and Specificity; Statistical analysis; Toxicology; Validation studies; Viability; Predictive capacity; MCTT HCE; PS; Statistical analysis; OECD TG; ROC; ICC; WL/BL; AUC; Eye irritation test; EIT; SD; GHS; Reproducibility; RhCE
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2016.11.010

Here, we report the results of a prevalidation trial for an in vitro eye irritation test (EIT) using the reconstructed human cornea-like epithelium, MCTT HCE™. The optimal cutoff to determine irritation in the prediction model was established at 35% with the receiver operation characteristics(ROC) curve for 126 substances. Within-lab(WL) and between-lab(BL) reproducibility was tested for 20 reference substances by 3 participating laboratories. Viability data described by mean±SD or ±1/2 difference between duplicate wells, and scatter plots, demonstrated the WL/BL consistency. WL/BL concordance with the binary decision, whether non-irritant or irritant was estimated to be 85–95% and 95%, respectively. WL/BL reproducibility of viability data was further supported by a strong correlation(ICC, r>0.9). WL/BL agreement of binary decisions was also examined by Fleiss' Kappa statistics, which showed a strong level of agreement (>0.78), nevertheless weaker than the reproducibility of the viability. The EIT with MCTT HCE™ exhibited a sensitivity of 82.2% (60/73), a specificity of 81.1% (43/53), and an accuracy of 81.8% (103/126) for 126 reference substances (for liquids; a sensitivity of 100% (47/47), a specificity of 70.6% (24/34), and an accuracy of 87.7% (71/81), and for solids, a sensitivity of 50% (13/26), a specificity of 100% (19/19), and an accuracy of 71.1% (32/45), suggesting that the accuracy is satisfactory but the sensitivity needs improvement, which shall be addressed through correcting the poor sensitivity for solid substances in future full validation trials. •MCTT™ HCE EIT employing water-soluble formazan WST-1, demonstrated a performance comparable to other RhCEs.•Within- and between-lab reproducibility for 3 labs with 20 chemicals were 85–95% and 95% respectively.•Predictive capacity for 126 chemicals was sensitivity 82.2%, specificity 81.1% and accuracy 81.8%