Abstract
Purpose :
To replace the Draize eye irritation test (OECD Test Guideline 404), several test methods based on reconstructed cornea-like epithelium (RhCE) have been developed and adopted in the OECD TG 492. The objective of this study was to define the essential structural, functional and procedural elements and to stablish the experimental procedures and evaluate the performance assessment of QobuR-RhCE, an in-house cruelty-free RhCE model to be used for the evaluation of eye hazard
Methods :
QobuR-RhCE consists in normal human corneal epithelial cells cultured at the air-liquid interface forming a stratified squamous epithelium resembling a native human corneal epithelium
Essential test method components consisting of structural, functional and procedural elements were evaluated and standardized. These components included unique characteristics of the test method (i.e. histology), critical procedural details (i.e. barrier function) and quality control measures
Performance assessment was evaluated in accordance with the revised performance standards for the assessment of proposed similar or modified in vitro reconstructed human cornea-like epithelium and the minimum list of reference chemicals was evaluated on QobuR-RhCE. In addition to standard cell viability evaluation, we describe a secondary endpoint based on Transepithelial Electrical Resistance (TEER) that could be of use to better discriminate between irritants and non-irritants
Accuracy, specificity, sensitivity and within-laboratory reproducibility (WLR) were calculated according to the OECD Performance Standards for the assessment of proposed similar or modified in vitro RhCE test methods for Eye Hazard
Results :
As result, the proposed method scored 93.3% sensibility, 60% specificity, 76.7% accuracy and 96.7% WLR, providing a similar performance in comparison to the validated reference methods. The inclusion of TEER as a barrier disruption predictor provided additional information for safety assessment that could be of interest during the evaluation of particular chemical products such as borderline chemical products, colored compounds or direct MTT reducers
Conclusions :
The results of this study suggest that QobuR-RhCE test method is capable of fulfilling the performance standards as described in OECD TG 492 for similar in vitro RhCE test methods for eye hazard, suggesting that QobuR-RhCE is a reliable method for predicting eye irritation
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.