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      Functional Evaluation of Two Corneal Endothelial Cell-Based Therapies: Tissue-Engineered Construct and Cell Injection

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          Abstract

          Restoration of vision due to corneal blindness from corneal endothelial dysfunction can be achieved via a corneal transplantation. However, global shortage of donor tissues has driven the development cell-based therapeutics. With the capacity to propagate regulatory compliant human corneal endothelial cells (CEnCs), this study evaluated the functionality of propagated CEnCs delivered via tissue-engineered endothelial keratoplasty (TE-EK) or corneal endothelial cell injection (CE-CI) within a rabbit model of bullous keratopathy. For animals with TE-EK grafts, central corneal thickness (CCT) increased to >1000 μm post-operatively. Gradual thinning with improvements in corneal clarity was observed from week 1. CCT at week 3 was 484.3 ± 73.7 μm. In rabbits with CE-CI, corneal clarity was maintained throughout, and CCT at week 3 was 582.5 ± 171.5 μm. Control corneas remained significantly edematous throughout the study period compared to their respective experimental groups ( p < 0.05). Characterization of excised corneas showed a monolayer with heterogeneously shaped CEnCs in both TE-EK and CE-CI groups. Immunohistochemistry demonstrated reactivity to anti-human specific nuclei antibody attributing corneal recovery to the functional human CEnCs. This study showed that regulatory compliant cell-based therapy for corneal endothelial dysfunction can be delivered by both TE-EK and CE-CI, and holds great promise as an alternative to traditional corneal transplantation.

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          Descemet-stripping automated endothelial keratoplasty.

          Alexey Gorovoy (2006)
          To evaluate the speed of visual recovery in 16 consecutive patients with corneal endothelial dysfunction who received Descemet-stripping automated endothelial keratoplasty (DSAEK). This is a retrospective study of a novel method for small-incision endothelial transplantation (DSAEK). Endothelial replacement was accomplished with Descemet stripping of the recipient and insertion of a posterior donor tissue that had been prepared with a microkeratome. Best spectacle-corrected visual acuity (BSCVA) by manifest refraction, endothelial counts, and dislocation rates were measured up to 12 months after DSAEK. Sixteen consecutive patients underwent uncomplicated DSAEK. Three patients had known optic nerve or macular disease precluding vision better than 20/200. Of the remaining 14 patients, 11 had BSCVA of 20/40 by postoperative week 12 (7 by week 6). The remaining 2 were 20/50 by weeks 6 and 12. All 14 patients were 20/40 or better at 1 year. One patient had a primary graft failure, and surgery was repeated with 20/40 BSCVA at 1 year. The dislocation rate was 25%. The average cell count between 7 and 10 months was 1714. The average pachymetry was 682. DSAEK surgery allows rapid, excellent BSCVA visual recovery. The rate of visual recovery is more rapid than usually found with penetrating keratoplasty.
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            Proliferative capacity of corneal endothelial cells.

            Jeffrey Joyce (2012)
            The corneal endothelial monolayer helps maintain corneal transparency through its barrier and ionic "pump" functions. This transparency function can become compromised, resulting in a critical loss in endothelial cell density (ECD), corneal edema, bullous keratopathy, and loss of visual acuity. Although penetrating keratoplasty and various forms of endothelial keratoplasty are capable of restoring corneal clarity, they can also have complications requiring re-grafting or other treatments. With the increasing worldwide shortage of donor corneas to be used for keratoplasty, there is a greater need to find new therapies to restore corneal clarity that is lost due to endothelial dysfunction. As a result, researchers have been exploring alternative approaches that could result in the in vivo induction of transient corneal endothelial cell division or the in vitro expansion of healthy endothelial cells for corneal bioengineering as treatments to increase ECD and restore visual acuity. This review presents current information regarding the ability of human corneal endothelial cells (HCEC) to divide as a basis for the development of new therapies. Information will be presented on the positive and negative regulation of the cell cycle as background for the studies to be discussed. Results of studies exploring the proliferative capacity of HCEC will be presented and specific conditions that affect the ability of HCEC to divide will be discussed. Methods that have been tested to induce transient proliferation of HCEC will also be presented. This review will discuss the effect of donor age and endothelial topography on relative proliferative capacity of HCEC, as well as explore the role of nuclear oxidative DNA damage in decreasing the relative proliferative capacity of HCEC. Finally, potential new research directions will be discussed that could take advantage of and/or improve the proliferative capacity of these physiologically important cells in order to develop new treatments to restore corneal clarity. Copyright © 2011 Elsevier Ltd. All rights reserved.
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              The corneal endothelium.

              D. Coster, S Tuft (1989)
              The endothelium is a monolayer of cells on the posterior corneal surface that transports water from the stroma into the anterior chamber. This movement of water counters a natural tendency for the stroma to swell and is necessary to maintain a transparent cornea. Embryologic studies, in particular the demonstration of the derivation of the endothelium from the neural crest, have provided insight into the factors that govern the response of this tissue to disease. In some species the endothelium can regenerate after injury, but in man cellular enlargement is the main mechanism of repair after cell loss. A clinical estimate of endothelial cell density and function is provided by specular microscopy, fluorophotometry and pachymetry. In this paper we review the development, structure and function of the corneal endothelium, and then consider the pathological processes that can affect this tissue.
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                Author and article information

                Contributors
                Gary S. L. Peh:
                ORCID: http://orcid.org/0000-0002-5387-4209
                garypeh@gmail.com
                Jodhbir S. Mehta: jodmehta@gmail.com
                Journal
                Journal ID (nlm-ta): Sci Rep
                Journal ID (iso-abbrev): Sci Rep
                Title: Scientific Reports
                Publisher: Nature Publishing Group UK (London )
                ISSN (Electronic): 2045-2322
                Publication date (Electronic): 15 April 2019
                Publication date PMC-release: 15 April 2019
                Publication date Collection: 2019
                Volume: 9
                Electronic Location Identifier: 6087
                Affiliations
                [1 ]ISNI 0000 0001 0706 4670, GRID grid.272555.2, Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, ; Singapore, Singapore
                [2 ]ISNI 0000 0004 0385 0924, GRID grid.428397.3, Duke-NUS Graduate Medical School, ; Singapore, Singapore
                [3 ]ISNI 0000 0000 9960 1711, GRID grid.419272.b, Singapore National Eye Centre, ; Singapore, Singapore
                [4 ]ISNI 0000 0001 2224 0361, GRID grid.59025.3b, School of Material Science and Engineering, , Nanyang Technological University, ; Singapore, Singapore
                Author information
                http://orcid.org/0000-0002-5387-4209
                http://orcid.org/0000-0001-5475-8712
                Article
                Publisher ID: 42493
                DOI: 10.1038/s41598-019-42493-3
                PMC ID: 6465252
                PubMed ID: 30988373
                SO-VID: 372344c6-daa7-4b47-91c0-be497d132666
                Copyright © © The Author(s) 2019
                License:

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                Date received : 22 June 2018
                Date accepted : 2 April 2019
                Funding
                Funded by: National Research Foundation Translational and Clinical Research (TCR) Programme (NMRC/TCR/008-SERI/2013)
                Categories
                Subject: Article
                Custom metadata
                issue-copyright-statement © The Author(s) 2019

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