Regulatory Compliant Tissue-Engineered Human Corneal Endothelial Grafts Restore Corneal Function of Rabbits with Bullous Keratopathy

Shu-Jun Lin,Maninder Bhogal,Khadijah Adnan,Chan N. Lwin,Xin-Yi Seah,Gary S. L. Peh,Benjamin L. George,Donald T. Tan,Heng-Pei Ang,Jodhbir S. Mehta,Yu-Chi Liu

doi:10.1038/s41598-017-14723-z

Abstract

Corneal transplantation is the only treatment available to restore vision for individuals with blindness due to corneal endothelial dysfunction. However, severe shortage of available donor corneas remains a global challenge. Functional regulatory compliant tissue-engineered corneal endothelial graft substitute can alleviate this reliance on cadaveric corneal graft material. Here, isolated primary human corneal endothelial cells (CEnCs) propagated using a dual media approach refined towards regulatory compliance showed expression of markers indicative of the human corneal endothelium, and can be tissue-engineered onto thin corneal stromal carriers. Both cellular function and clinical adaptability was demonstrated in a pre-clinical rabbit model of bullous keratopathy using a tissue-engineered endothelial keratoplasty (TE-EK) approach, adapted from routine endothelial keratoplasty procedure for corneal transplantation in human patients. Cornea thickness of rabbits receiving TE-EK graft gradually reduced over the first two weeks, and completely recovered to a thickness of approximately 400 µm by the third week of transplantation, whereas corneas of control rabbits remained significantly thicker over 1,000 µm (p < 0.05) throughout the course of the study. This study showed convincing evidence of the adaptability of the propagated CEnCs and their functionality via a TE-EK approach, which holds great promises in translating the use of cultured CEnCs into the clinic.

Highlights

Performed in 116 countries in 2012 alone, but a conservative estimate made in the same reports puts demand for corneal transplantation at around 12.7 million[7], providing the heightened impetus for the development of alternate treatment strategies through scalable cell-based regenerative therapeutics
We have described an approach for the isolation and propagation of primary human corneal endothelial cells (CEnCs) using a robust dual media culture system, where the isolated CEnCs were cultured in a proliferative medium until they are near confluence before being switched into a maintenance medium[11]
Even though 184,576 corneal transplants were performed in 2012 worldwide[7], there is a global shortage of donor corneas far in excess of this number, and the demand for corneal transplantation will undoubtedly increase with an aging global population[31]

Summary

Introduction

Performed in 116 countries in 2012 alone, but a conservative estimate made in the same reports puts demand for corneal transplantation at around 12.7 million[7], providing the heightened impetus for the development of alternate treatment strategies through scalable cell-based regenerative therapeutics. We have described an approach for the isolation and propagation of primary human CEnCs using a robust dual media culture system, where the isolated CEnCs were cultured in a proliferative medium until they are near confluence before being switched into a maintenance medium[11]. From start to finish, this study shows the robustness and adaptability of propagating human CEnCs using the dual media approach, and most importantly, provides an insight into the functional translation of tissue-engineered graft material generated using these CEnCs for the replacement of a damaged CE layer through current corneal transplant surgical technique

Methods

Results

Conclusion