Characterization of tissue-engineered posterior corneas using second- and third-harmonic generation microscopy.
Three-dimensional tissues, such as the cornea, are now being engineered as substitutes for the rehabilitation of vision in patients with blinding corneal diseases. Engineering of tissues for translational purposes requires a non-invasive monitoring to control the quality of the resulting biomaterial. Unfortunately, most current methods still imply invasive steps, such as fixation and staining, to clearly observe the tissue-engineered cornea, a transparent tissue with weak natural contrast. Second- and third-harmonic generation imaging are well known to provide high-contrast, high spatial resolution images of such tissues, by taking advantage of the endogenous contrast agents of the tissue itself. In this article, we imaged tissue-engineered corneal substitutes using both harmonic microscopy and classic histopathology techniques. We demonstrate that second- and third-harmonic imaging can non-invasively provide important information regarding the quality and the integrity of these partial-thickness posterior corneal substitutes (observation of collagen network, fibroblasts and endothelial cells). These two nonlinear imaging modalities offer the new opportunity of monitoring the engineered corneas during the entire process of production.
Centre Énergie Matériaux Télécommunications, Institut National de la Recherche Scientifique, Varennes, Quebec, Canada; Centre de recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Quebec, Canada and Département d'ophtalmologie, Université de Montréal, Montréal, Quebec, Canada.
Louis
L
Jay
Centre de recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Quebec, Canada and Département d'ophtalmologie, Université de Montréal, Montréal, Quebec, Canada.
Jean-Michel
JM
Bourget
Axe médecine régénératrice, Hôpital du Saint-Sacrement, Centre de recherche du CHU de Québec, Québec, Quebec, Canada and Centre de recherche en organogénèse expérimentale de l'Université Laval / LOEX, Québec, Quebec, Canada.
Benjamin
B
Goyer
Centre Énergie Matériaux Télécommunications, Institut National de la Recherche Scientifique, Varennes, Quebec, Canada; Centre de recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Quebec, Canada and Département d'ophtalmologie, Université de Montréal, Montréal, Quebec, Canada.
Kanwarpal
K
Singh
Centre de recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, Quebec, Canada and Département d'ophtalmologie, Université de Montréal, Montréal, Quebec, Canada.
Isabelle
I
Brunette
Centre Énergie Matériaux Télécommunications, Institut National de la Recherche Scientifique, Varennes, Quebec, Canada.
Tsuneyuki
T
Ozaki
Axe médecine régénératrice, Hôpital du Saint-Sacrement, Centre de recherche du CHU de Québec, Québec, Quebec, Canada and Centre de recherche en organogénèse expérimentale de l'Université Laval / LOEX, Québec, Quebec, Canada; Département d'ophtalmologie et d'oto-rhino-laryngologie, Faculté de médecine, Université Laval, Québec, Quebec, Canada.
Stéphanie
S
Proulx
D000328:Adult; D003094:Collagen / Q000378:metabolism; D003315:Cornea / Q000502:physiology*; D003319:Corneal Stroma / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D004728:Endothelium, Corneal / Q000166:cytology; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008853:Microscopy / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods*
D003094:Collagen