title: Characterization of a corneal endothelium engineered on a self-assembled stromal substitute.
abstract: Endothelial dysfunctions are the first indication for allogeneic corneal transplantation. Development of a tissue-engineered posterior cornea could be an alternative to the use of native allogeneic tissues. In this paper, we used the self-assembly approach to form a cellularized stromal substitute that served as a carrier for the engineering of an endothelium. This endothelialized stromal substitute was then characterized using alizarin red staining, histology, scanning and transmission electron microscopy, as well as mass spectrometry and immunodetection of collagens and function-related proteins. We report the engineering of a monolayer of flattened endothelial cells with a cell density of 966 ± 242 cells/mm(2) (mean ± SD). Endothelial interdigitations were present between cells. The stromal fibroblasts deposited a dense and cohesive collagenous matrix. Collagen fibrils had a diameter of 39.1 ± 11.3 nm, and a mean center to center interfibrillar space of 50.9 ± 10.9 nm. The stromal substitute was composed of collagen types I, V, VI and XII, as well as lumican and decorin. Type IV collagen was also present underneath the endothelium. The endothelium expressed both the sodium/potassium (Na(+)/K(-)) ATPase and sodium/bicarbonate (Na(+)/ [Formula: see text] ) cotransporter pumps. These results indicate that the self-assembled stromal substitute is able to support the expression of endothelial cell functionality markers and therefore, is a suitable carrier for the engineering of an endothelium that could be used for the treatment of endothelial dysfunctions.
authors.affiliation: Centre de recherche du Centre hospitalier universitaire (CHU) de Québec, Université Laval, axe médecine régénératrice, Hôpital du Saint-Sacrement, 1050 chemin Ste-Foy, G1S 4L8, Québec, QC, Canada; Centre de recherche de l'hôpital Maisonneuve-Rosemont, 5415 boulevard de l'Assomption, H1T 2M4, Montréal, QC, Canada; Département d'ophtalmologie, Université de Montréal, Montréal, QC, Canada. Electronic address: Jean-michel.bourget.1@ulaval.ca.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Bourget
authors.affiliation: Centre de recherche du Centre hospitalier universitaire (CHU) de Québec, Université Laval, axe médecine régénératrice, Hôpital du Saint-Sacrement, 1050 chemin Ste-Foy, G1S 4L8, Québec, QC, Canada; Département d'Ophtalmologie, Faculté de médecine, Université Laval, Québec, QC, Canada; Centre de recherche en organogénèse expérimentale de l'Université Laval, LOEX, 1401 18(ème) rue, G1J 1Z4, Québec, QC, Canada. Electronic address: Stephanie.Proulx@fmed.ulaval.ca.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Proulx
mesh_terms: D000818:Animals; D016039:Corneal Transplantation; D004728:Endothelium, Corneal / Q000637:transplantation / Q000648:ultrastructure*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D023822:Tissue Engineering / Q000379:methods*
keywords: Corneal endothelium; Endothelial cells; Living stromal carrier; Posterior cornea; Self-assembly; Tissue engineering