title: In Vivo Functionality of a Corneal Endothelium Transplanted by Cell-Injection Therapy in a Feline Model.
abstract: OBJECTIVE
  To evaluate the functionality of a corneal endothelium reconstituted by injection of corneal endothelial cells (CEC) in the anterior chamber of a feline model.
  
  
  METHODS
  We operated the right eyes of 16 animals. Eight underwent central endothelial scraping and injection with 2 × 10(5) (n = 4) or 1 × 10(6) (n = 4) feline CEC supplemented with Y-27632 and labeled with 3,3'-Dioctadecyl-5,5'-Di(4-Sulfophenyl)Oxacarbocyanine (SP-DiOC18[3] or DiOC). After total endothelial scraping, two eyes were injected with 1 × 10(6) 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.
  
  
  RESULTS
  Postoperatively, corneas grafted with 2 × 10(5) CEC and centrally scraped controls displayed the best transparency and pachymetry. Corneas grafted with 1 × 10(6) 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 × 10(5) CEC and centrally scraped controls, a nonuniform endothelial multilayer without expression of functional proteins in centrally scraped corneas grafted with 1 × 10(6) CEC, and a nonfunctional fibrotic endothelium in entirely scraped grafts and controls. Expression of DiOC in grafts was scarce.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Maisonneuve-Rosemont Hospital Research Center, Montreal, Quebec, Canada 2Department of Ophthalmology, University of Montreal, Montreal, Quebec, Canada.
authors.forename: Cristina
authors.initials: C
authors.lastname: Bostan
authors.affiliation: Centre d'organogénèse expérimentale de l'Université Laval/LOEX, Québec City, Quebec, Canada, and Centre de recherche du CHU de Québec-UL, Axe Médecine régénératrice, Québec City, Quebec, Canada.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Thériault
authors.affiliation: Maisonneuve-Rosemont Hospital Research Center, Montreal, Quebec, Canada.
authors.forename: Karolyn J
authors.initials: KJ
authors.lastname: Forget
authors.affiliation: Maisonneuve-Rosemont Hospital Research Center, Montreal, Quebec, Canada 2Department of Ophthalmology, University of Montreal, Montreal, Quebec, Canada.
authors.forename: Christelle
authors.initials: C
authors.lastname: Doyon
authors.affiliation: Ophthalmology and Visual Neurosciences and Laboratory Medicine and Pathology, University of Minnesota School of Medicine, Minneapolis, Minnesota, United States.
authors.forename: J Douglas
authors.initials: JD
authors.lastname: Cameron
authors.affiliation: Centre d'organogénèse expérimentale de l'Université Laval/LOEX, Québec City, Quebec, Canada, and Centre de recherche du CHU de Québec-UL, Axe Médecine régénératrice, Québec City, Quebec, Canada 5Department of Ophthalmology and ENT-Head and Neck Surgery, U.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Proulx
authors.affiliation: Maisonneuve-Rosemont Hospital Research Center, Montreal, Quebec, Canada 2Department of Ophthalmology, University of Montreal, Montreal, Quebec, Canada.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Brunette
mesh_terms: D000577:Amides / Q000494:pharmacology; D000818:Animals; D002415:Cats; D002452:Cell Count; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D063171:Corneal Pachymetry; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000502:physiology* / Q000637:transplantation*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006085:Graft Survival / Q000502:physiology; D056965:Injections, Intraocular; D008854:Microscopy, Electron; D011725:Pyridines / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014184:Transplantation, Homologous; D014945:Wound Healing / Q000502:physiology; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D000577:Amides; D004791:Enzyme Inhibitors; D011725:Pyridines; D062826:Zonula Occludens-1 Protein; C108830:Y 27632; D000254:Sodium-Potassium-Exchanging ATPase