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

Agnė Žiniauskaitė,Romuald Eimont,Artūras Ulčinas,Jenni J Hakkarainen,Ramūnas Valiokas,Rūta Aldonytė,Vytautas Cėpla,Tadas Jelinskas,Giedrius Kalesnykas

doi:10.3390/sci3030030

Abstract

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 (Papp) values of a representative set of ophthalmic drugs were measured and correlated to rabbit cornea Papp 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.

Highlights

Introduction published maps and institutional affilThe topical administration of ophthalmic drugs remains the preferred route of drug delivery to ocular tissues
In this paper, we presented an in vitro corneal mimetic where a chemically crosslinked collagen-based, self-supporting hydrogel is employed as a matrix for a human corneal epithelial multilayer
We report on the engineering of a human cornea-mimicking device compatible with established drug permeability analysis techniques

Summary

Introduction

Introduction published maps and institutional affilThe topical administration of ophthalmic drugs remains the preferred route of drug delivery to ocular tissues. The superficial apical layer of the epithelial cells contains tight junctions that seal the intercellular spaces between them, making the corneal epithelium into an efficient diffusion barrier. The corneal epithelium represents a ratelimiting barrier for topically administered hydrophilic drugs and macromolecules. The corneal stroma consists mostly of water (~80%) and type I collagen fibrils and makes up to 90% of the thickness of the cornea [1,2]. With these properties, the corneal stroma restricts the penetration of hydrophobic drug molecules effectively [1]

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