Introducing an Efficient In Vitro Cornea Mimetic Model for Testing Drug Permeability

• Main Authors: Agnė Žiniauskaitė, Vytautas Cėpla, Tadas Jelinskas, Romuald Eimont, Artūras Ulčinas, Rūta Aldonytė, Ramūnas Valiokas, Giedrius Kalesnykas, Jenni J. Hakkarainen
• Format: Article
• Language: English
• Published: MDPI AG 2021-06-01
• Series: Sci
• Subjects: in vitro corneal model; collagen hydrogel; permeability
• Online Access: https://www.mdpi.com/2413-4155/3/3/30

There is a growing need for novel in vitro corneal models to replace animal-based ex vivo tests in drug permeability studies. In this study, we demonstrated a corneal mimetic that models the stromal and epithelial compartments of the human cornea. Human corneal epithelial cells (HCE-T) were grown on top of a self-supporting porcine collagen-based hydrogel. Cross-sections of the multi-layers were characterized by histological staining and immunocytochemistry of zonula oc-cludens-1 protein (ZO-1) and occludin. Furthermore, water content and bssic elastic properties of the synthetized collagen type I-based hydrogels were measured. The apparent permeability coefficient (P_app) values of a representative set of ophthalmic drugs were measured and correlated to rabbit cornea P_app values found in the literature. A multilayered structure of HCE-T cells and the expression of ZO-1 and occludin in the full thickness of the multilayer were observed. The hydrogel-based corneal model exhibited an excellent correlation to rabbit corneal permeability (r = 0.96), whereas the insert-grown HCE-T multilayer was more permeable and the correlation to the rabbit corneal permeability was lower (r = 0.89). The hydrogel-based human corneal model predicts the rabbit corneal permeability more reliably in comparison to HCE-T cells grown in inserts. This in vitro human corneal model can be successfully employed for drug permeability tests whilst avoiding ethical issues and reducing costs.