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Originally published In Press as doi:10.1167/iovs.08-2793 on January 17, 2009
 (Investigative Ophthalmology and Visual Science. 2009;50:2686-2694.)
© 2009 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.08-2793
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Transplantation of a Tissue-Engineered Corneal Endothelium Reconstructed on a Devitalized Carrier in the Feline Model

Stéphanie Proulx,1,2,3 Thouria Bensaoula,4 Ossama Nada,4 Caroline Audet,1,2,3 Jeanne d'Arc Uwamaliya,1,2,3 Angèle Devaux,5 Guy Allaire,5,6 Lucie Germain,1,2,3 and Isabelle Brunette4,5

1From the Laboratory of Experimental Organogenesis, CHA Universitaire de Québec, Quebec, QC, Canada; the Departments of 2Oto-Rhino-Laryngology and Ophthalmology and 3Surgery, Laval University, Quebec, QC, Canada; 4Maisonneuve-Rosemont Hospital Research Center, Montreal, QC, Canada; the 5Department of Ophthalmology, University of Montreal, Montreal, QC, Canada; and the 6Department of Pathology, CHUM-Notre Dame, Montreal, QC, Canada.

PURPOSE. To evaluate the functional outcome of tissue-engineered corneal endothelium reconstructed on a devitalized carrier and transplanted in the living feline model.

METHODS. Eighteen healthy adult cats underwent full-thickness corneal transplantation. In 11 animals, the donor cornea was reconstructed from cultured allogeneic feline corneal endothelial cells seeded on the denuded Descemet’s membrane of a devitalized human cornea. The reconstructed corneal endothelium was cultured for 2 weeks before transplantation. Five control animals received autologous (n = 1), allogeneic (n = 3), or human xenogeneic (n = 1) native cornea. Two other control animals were grafted with the devitalized carrier only (no cells). Animals were observed daily by slit lamp until euthanatization on day 7. Postmortem analysis included optical coherence tomography (OCT), alizarin red staining, histology, fluorescence microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM).

RESULTS. Nine of the 11 reconstructed corneal endothelial grafts and all five native (autologous, allogeneic, xenogeneic) control grafts were clear and thin 7 days after grafting. In contrast, the two control grafts consisting of the carrier only (without endothelium) remained thick and opaque. Alizarin red staining, histology, SEM, and TEM showed that the transplanted reconstructed endothelium maintained a normal morphology and ultrastructure and expressed the function-related proteins Na+/K+-ATPase {alpha}1, Na+/HCO3, and ZO-1.

CONCLUSIONS. This study provides evidence for the short-term (7-day) anatomic and functional success of corneal transplantation with a tissue-engineered corneal endothelium reconstructed on a devitalized carrier.






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