Transplantation of a tissue-engineered corneal endothelium reconstructed on a devitalized carrier in the feline model.
OBJECTIVE
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 alpha1, Na(+)/HCO(3), 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.
Proulx
Stéphanie
S
Laboratory of Experimental Organogenesis, CHA Universitaire de Québec, Quebec, QC, Canada.
Bensaoula
Thouria
T
Nada
Ossama
O
Audet
Caroline
C
d'Arc Uwamaliya
Jeanne
J
Devaux
Angèle
A
Allaire
Guy
G
Germain
Lucie
L
Brunette
Isabelle
I
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002415:Cats; D002452:Cell Count; D002478:Cells, Cultured; D003319:Corneal Stroma / Q000166:cytology; D016039:Corneal Transplantation; D003886:Descemet Membrane / Q000166:cytology; D004728:Endothelium, Corneal / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010750:Phosphoproteins / Q000378:metabolism; D027982:Sodium-Bicarbonate Symporters / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D041623:Tomography, Optical Coherence; D014182:Transplantation, Autologous; D014183:Transplantation, Heterologous; D014184:Transplantation, Homologous; D062826:Zonula Occludens-1 Protein
D008565:Membrane Proteins; D010750:Phosphoproteins; D027982:Sodium-Bicarbonate Symporters; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D000254:Sodium-Potassium-Exchanging ATPase