Development and Validation of an In Vitro Ocular Irritation Test for Ophthalmic Medical Devices with a Novel Reconstructed Human Corneal Epithelium Model

For testing ocular irritation, 3D corneal models mimicking the corneal epithelium are considered reliable eye irritation tests and are detailed in regulatory guideline OECD Test Guideline (TG) 492. The aim of the present study was to develop and validate a Reconstructed human Cornea-like Epithelium (RhCE) in vitro irritation test method for ophthalmic medical devices according to OECD TG 492. Immortalized Human Primary Corneal Epithelium Cells (IM-HCEpiCs) were cultured on microporous inserts and exposed to an Air-Liquid Interface (ALI). Morphology was examined using standard (immuno-) histological techniques. Viability was quantified with MTT assay. Barrier integrity and function were monitored by trans-epithelial electrical resistance (TEER) and determination of IC50 using MTT assay. Reproducibility was evaluated by calculating the inter-batch coefficient of variation (CV %) of the absorbance values of negative control-treated RhCE model replicates by MTT assay. Technical proficiency was verified using reference chemicals. Irritancy of ophthalmic medical devices was assessed. IM-HCEpiCs developed an epithelium-like barrier under the ALI. TEER increased after ALI introduction, and the obtained IC50 value showed concordance with the guideline's reference ranges. The developed RhCE test method demonstrated technical proficiency and correctly identified medical devices as non-irritants. A novel RhCE model was developed and validated according to OECD TG 492.