{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T08:01:14Z","timestamp":1781164874594,"version":"3.54.1"},"reference-count":33,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T00:00:00Z","timestamp":1778457600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>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\u2013Liquid 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\u2019s 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.<\/jats:p>","DOI":"10.3390\/bioengineering13050545","type":"journal-article","created":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T16:07:26Z","timestamp":1779206846000},"page":"545","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Validation of an In Vitro Ocular Irritation Test for Ophthalmic Medical Devices with a Novel Reconstructed Human Corneal Epithelium Model"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-7271-2758","authenticated-orcid":false,"given":"Payal","family":"Rawat","sequence":"first","affiliation":[{"name":"Research and Development, AL.CHI.MI.A. S.R.L., Moria Group, 35020 Ponte San Nicol\u00f2, Italy"},{"name":"Department of Biology, University of Padova, 35131 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4910-1617","authenticated-orcid":false,"given":"Umberto","family":"Rodella","sequence":"additional","affiliation":[{"name":"Research and Development, AL.CHI.MI.A. S.R.L., Moria Group, 35020 Ponte San Nicol\u00f2, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6938-6851","authenticated-orcid":false,"given":"Stefania","family":"D\u2019Agostino","sequence":"additional","affiliation":[{"name":"Fondazione Banca Degli Occhi del Veneto ETS (FBOV), 30174 Venice, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0390-6823","authenticated-orcid":false,"given":"Eugenio","family":"Ragazzi","sequence":"additional","affiliation":[{"name":"Studium Patavinum, University of Padova, 35122 Padova, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0760-3723","authenticated-orcid":false,"given":"Orietta","family":"Rossi","sequence":"additional","affiliation":[{"name":"Quality Control, AL.CHI.MI.A. S.R.L., Moria Group, 35020 Ponte San Nicol\u00f2, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4960-3614","authenticated-orcid":false,"given":"Claudio","family":"Gatto","sequence":"additional","affiliation":[{"name":"Research and Development, AL.CHI.MI.A. S.R.L., Moria Group, 35020 Ponte San Nicol\u00f2, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1949-5213","authenticated-orcid":false,"given":"Laura","family":"Giurgola","sequence":"additional","affiliation":[{"name":"Research and Development, AL.CHI.MI.A. S.R.L., Moria Group, 35020 Ponte San Nicol\u00f2, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1498-615X","authenticated-orcid":false,"given":"Jana","family":"D\u2019Amato T\u00f3thov\u00e1","sequence":"additional","affiliation":[{"name":"Research and Development, AL.CHI.MI.A. S.R.L., Moria Group, 35020 Ponte San Nicol\u00f2, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1002\/jbm.a.31619","article-title":"A methodology based on the \u201canterior chamber of rabbit eyes\u201d model for noninvasively determining the biocompatibility of biomaterials in an immune privileged site","volume":"86A","author":"Lu","year":"2008","journal-title":"J. Biomed. Mater. Res. Part A"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"105371","DOI":"10.1016\/j.tiv.2022.105371","article-title":"ISO 10993-23 In vitro irritation testing for medical devices: Substantiating applicability to mild irritants and non-extractables","volume":"82","author":"Pellevoisin","year":"2022","journal-title":"Toxicol. In Vitro"},{"key":"ref_3","unstructured":"Ludwig, P.E., Lopez, M.J., and Sevensma, K.E. (2025, December 31). 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