Development of human corneal epithelium on organized fibrillated transparent collagen matrices synthesized at high concentration.
Several diseases can lead to opacification of cornea requiring transplantation of donor tissue to restore vision. In this context, transparent collagen I fibrillated matrices have been synthesized at 15, 30, 60 and 90 mg/mL. The matrices were evaluated for fibril organizations, transparency, mechanical properties and ability to support corneal epithelial cell culture. The best results were obtained with 90 mg/mL scaffolds. At this concentration, the fibril organization presented some similarities to that found in corneal stroma. Matrices had a mean Young's modulus of 570 kPa and acellular scaffolds had a transparency of 87% in the 380-780 nm wavelength range. Human corneal epithelial cells successfully colonized the surface of the scaffolds and generated an epithelium with characteristics of corneal epithelial cells (i.e. expression of cytokeratin 3 and presence of desmosomes) and maintenance of stemness during culture (i.e. expression of ΔNp63α and formation of holoclones in colony formation assay). Presence of cultured epithelium on the matrices was associated with increased transparency (89%).
Sorbonne Universités, UPMC Univ Paris 06, CNRS, Collège de France, UMR 7574, Chimie de la Matière Condensée de Paris, F-75005 Paris, France.
Aurélien
A
Tidu
Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F75012, France; INSERM, U968, Paris F75012, France; CNRS, UMR_7210, Paris F75012, France; Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, DHU View Maintain, INSERM-DHOS CIC 1423, Paris F-75012, France.
Djida
D
Ghoubay-Benallaoua
Solids Mechanics Laboratory, Ecole Polytechnique, Centre National de la Recherche Scientifique, Palaiseau, France.
Barbara
B
Lynch
Sorbonne Universités, UPMC Univ Paris 06, CNRS, Collège de France, UMR 7574, Chimie de la Matière Condensée de Paris, F-75005 Paris, France.
Bernard
B
Haye
Sorbonne Universités, UPMC Univ Paris 06, CNRS, Collège de France, UMR 7574, Chimie de la Matière Condensée de Paris, F-75005 Paris, France.
Corinne
C
Illoul
Solids Mechanics Laboratory, Ecole Polytechnique, Centre National de la Recherche Scientifique, Palaiseau, France.
Jean-Marc
JM
Allain
Sorbonne Universités, UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F75012, France; INSERM, U968, Paris F75012, France; CNRS, UMR_7210, Paris F75012, France; Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, DHU View Maintain, INSERM-DHOS CIC 1423, Paris F-75012, France.
Vincent M
VM
Borderie
Sorbonne Universités, UPMC Univ Paris 06, CNRS, Collège de France, UMR 7574, Chimie de la Matière Condensée de Paris, F-75005 Paris, France. Electronic address: Gervaise.Mosser@upmc.fr.
Gervaise
G
Mosser
D016475:3T3 Cells; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D019573:Epithelium, Corneal / Q000166:cytology*; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D024022:Fibrillar Collagens / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D008422:Materials Testing; D051379:Mice; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods*
D024022:Fibrillar Collagens
Collagen; Human corneal epithelium; Limbal stem cell; Liquid-crystal; Transparent matrices