A new corneal epithelial biomimetic 3D model for in vitro eye toxicity assessment: Development, characterization and applicability

• Published in: Toxicology in vitro 2020-02, Vol.62, p.104666, Article 104666
• Main Authors: da Silva, Artur C.G., Chialchia, Adrienny R., de Castro, Elisandra G., e Silva, Marlos R.L., Arantes, Diego A.C., Batista, Aline C., Kitten, Gregory T., Valadares, Marize C.
• Format: Article
• Language: English
• Subjects: 3D corneal epithelial model; Animal Testing Alternatives; Biomarkers; Biomimetics; CD44 antigen; Cell Culture Techniques; Cell Line; Cell lineage; Chemicals; Classification; Cornea; Cultivation; Cytokeratin; Cytology; Epithelium, Corneal - drug effects; Evaluation; Experimentation; Eye irritation assessment; Eyebrow henna; Human corneal epithelium; Human tissues; Humans; Irritants - toxicity; Irritation; Models, Biological; Morphology; Organic chemistry; Three dimensional models; Tissues; Toxicity; Toxicity Tests - methods; Viability
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2019.104666

In vitro eye toxicity assessment using reconstructed corneal epithelial models has emerged highlighting its applicability domain for Classification and Labeling of products and chemicals. However, due to bureaucratic issues, such models are not commercially available in Brazil and Latin America. In this work, we developed, characterized and evaluated the applicability of a new corneal epithelial biomimetic model using a cell lineage for in vitro eye toxicity assessment. The reconstructed tissue was obtained through the cultivation of HaCaT cells in an air-liquid interface, which presented morphology and biomarkers expression such as cytokeratin, CD44, and Ki-67 similar to human tissue. Furthermore, tissue viability was evaluated after exposure of the epithelial model to isolated chemicals from different Globally Harmonized System (GHS) eye irritation categories, and it has been demonstrated to be a suitable endpoint for classification of test materials, allowing discrimination between irritant and non-irritant chemicals. Furthermore, the model showed suitability for testing “real-life mixtures”, once it identified irritant products between the analyzed eyebrow henna samples commercially labeled as non-irritants. This reproducible and low-cost epithelial corneal model presents features very important for Brazil and South America for R&D&I with no unnecessary animal experimentation. •A new corneal epithelial biomimetic model was reconstructed using the HaCaT human keratinocyte cell line.•The obtained tissue resembles the human corneal epithelium regarding tissue morphology and biomarkers expression.•The proposed model allowed differentiation between eye irritant and non-irritant isolated chemicals.•The epithelial model allowed eye toxicity assessment of “real-life mixtures”, represented by eyebrow henna samples.