Treatment of corneal endothelial damage in a rabbit model with a bioengineered graft using human decellularized corneal lamina and cultured human corneal endothelium.

Francisco Arnalich-Montiel,Marta Cadenas Martin,Sherezade Fuentes-Julián,Ane Pérez-Sarriegui,Gary Peh,Khadijah Adnan,Jodhbir S Mehta,Adrian Moratilla,Laura Porrua,Maria P De Miguel,Veronica Aparicio,Che J Connon

doi:10.1371/journal.pone.0225480

Abstract

We aimed to investigate the functionality of human decellularized stromal laminas seeded with cultured human corneal endothelial cells as a tissue engineered endothelial graft (TEEK) construct to perform endothelial keratoplasty in an animal model of corneal endothelial damage. Engineered corneal endothelial grafts were constructed by seeding cultured human corneal endothelial cell (hCEC) suspensions onto decellularized human corneal stromal laminas with various coatings. The functionality and survival of these grafts with cultured hCECs was examined in a rabbit model of corneal endothelial damage after central descemetorhexis. Rabbits received laminas with and without hCECs (TEEK and control group, respectively). hCEC seeding over fibronectin-coated laminas provided an optimal and consistent endothelial cell count density and polygonal shape on the decellularized laminas, showing active pump fuction. Surgery was performed uneventfully as standard Descemet stripping automated endothelial keratoplasty (DSAEK). Corneal transparency gradually recovered in the TEEK group, whereas haze and edema persisted for up to 4 weeks in the controls. Histologic examination showed endothelial cells of human origin covering the posterior surface of the graft in the TEEK group. Grafting of decellularized stroma carriers re-surfaced with human corneal endothelial cells ex vivo can be a readily translatable method to improve visual quality in corneal endothelial diseases.

Highlights

Corneas are the most commonly transplanted tissue worldwide; in 2012, 184,577 corneal transplants were performed in 116 countries but tissue was only procured in 82 countries [1]
Grafting of decellularized stroma carriers re-surfaced with human corneal endothelial cells ex vivo can be a readily translatable method to improve visual quality in corneal endothelial diseases
The FNC coating maintained the human corneal endothelial cell (hCEC) natural flat morphology; their embedding into the lamina collagen fibers could be observed, and it showed the best adhesion and morphology (Fig 2B)

Summary

Introduction

Corneas are the most commonly transplanted tissue worldwide; in 2012, 184,577 corneal transplants were performed in 116 countries but tissue was only procured in 82 countries [1]. Despite these high numbers, eye banks cannot match the demand worldwide and it is estimated that there is 1 cornea available per 70 needed [1]. Corneal bioengineering using expanded human corneal endothelial cells (hCECs) appears to be a feasible and viable technique in the short term for supplying extra tissue for endothelial keratoplasty (EK)[4,5]. Human endothelial cells are quiescent in vivo, and had proven extremely difficult to expand in vitro until recently [6,7,8,9,10,11]

Methods

Results

Conclusion