Establishment of an Ex Vivo Human Corneal Endothelium Wound Model.

Abstract

Purpose: A human model able to simulate the manifestation of corneal endothelium decompensation could be advantageous for wound healing and future cell therapy assessment. The study aimed to establish an ex vivo human cornea endothelium wound model where endothelium function can be evaluated by measuring corneal thickness changes.

Methods: The human cornea was maintained in an artificial anterior chamber, with a continuous culture medium infusion system designed to sustain corneal endothelium and epithelium simultaneously. The corneal thickness was used to assess corneal endothelial cell function. Immunostaining was used to evaluate cell viability and endothelial cell marker expression, ZO-1 and Na/K ATPase.

Results: Human corneas with intact corneal endothelium were maintained in the ex vivo model for 28 days, showing normal corneal thickness with a clear and transparent appearance. Corneal endothelial cells were alive and expressed ZO-1 and Na/K ATPase at the end of the organ culture. The endothelium wounded corneas showed persistent corneal edema with an increase in corneal thickness at 654.6 ± 31.7 µm. Staining results showed that no cells migrated to cover the wound and no expression of ZO-1 and Na/K ATPase on the posterior surface of the cornea was found.

Conclusions: This study provided a novel method to establish an ex vivo human cornea organ culture model, where corneal endothelium function can be evaluated by accessing the corneal thickness.

Translational relevance: The ex vivo model established in this study can provide an alternative to the animal model in studying corneal endothelium decompensation.