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Cristina Bostan, Mathieu Thériault, Karolyn J. Forget, Christelle Doyon, J. Douglas Cameron, Stéphanie Proulx, Isabelle Brunette; In Vivo Functionality of a Corneal Endothelium Transplanted by Cell-Injection Therapy in a Feline Model. Invest. Ophthalmol. Vis. Sci. 2016;57(4):1620-1634. doi: https://doi.org/10.1167/iovs.15-17625.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the functionality of a corneal endothelium reconstituted by injection of corneal endothelial cells (CEC) in the anterior chamber of a feline model.
We operated the right eyes of 16 animals. Eight underwent central endothelial scraping and injection with 2 × 105 (n = 4) or 1 × 106 (n = 4) feline CEC supplemented with Y-27632 and labeled with 3,3′-Dioctadecyl-5,5′-Di(4-Sulfophenyl)Oxacarbocyanine (SP-DiOC18 or DiOC). After total endothelial scraping, two eyes were injected with 1 × 106 labeled CEC and Y-27632. The central (n = 3) or entire (n = 3) endothelium was scraped in six eyes followed by Y-27632 injection without CEC. Subjects were positioned eyes down for 3 hours. Outcomes included graft transparency, pachymetry, CEC morphometry, histology, electron microscopy, and function and wound healing–related protein immunostaining.
Postoperatively, corneas grafted with 2 × 105 CEC and centrally scraped controls displayed the best transparency and pachymetry. Corneas grafted with 1 × 106 CEC yielded intermediate results. Entirely scraped controls remained hazy and thick. Histopathology revealed a confluent endothelial monolayer expressing sodium-potassium adenosine triphosphatase (Na+/K+-ATPase) and zonula occludens-1 (ZO-1) in corneas grafted with 2 × 105 CEC and centrally scraped controls, a nonuniform endothelial multilayer without expression of functional proteins in centrally scraped corneas grafted with 1 × 106 CEC, and a nonfunctional fibrotic endothelium in entirely scraped grafts and controls. Expression of DiOC in grafts was scarce.
Injected CEC contributed little to the incompletely functional endothelium of grafted corneas. Y-27632 injection without CEC following scraping reconstituted the healthiest endothelium. Further studies investigating the therapeutic effect of Y-27632 alone are needed to validate these conclusions.
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