Development of human corneal epithelium on organized fibrillated transparent collagen matrices synthesized at high concentration

• Published in: Acta biomaterialia 2015-08, Vol.22, p.50-58
• Main Authors: Tidu, Aurélien, Ghoubay-Benallaoua, Djida, Lynch, Barbara, Haye, Bernard, Illoul, Corinne, Allain, Jean-Marc, Borderie, Vincent M., Mosser, Gervaise
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Aged; Aged, 80 and over; Animals; Biochemistry, Molecular Biology; Cells, Cultured; Chemical Sciences; Collagen; Collagens; Diseases; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - ultrastructure; Epithelium; Epithelium, Corneal - cytology; Extracellular Matrix - drug effects; Extracellular Matrix - metabolism; Extracellular Matrix - ultrastructure; Fibrillar Collagens - pharmacology; Formations; Human; Human corneal epithelium; Humans; Immunohistochemistry; Life Sciences; Limbal stem cell; Liquid-crystal; Material chemistry; Materials Testing; Mice; Organizations; Rats, Sprague-Dawley; Rats, Wistar; Real-Time Polymerase Chain Reaction; Restoration; Scaffolds; Tissue Engineering - methods; Transparent matrices
• ISSN: 1742-7061; 1878-7568
• DOI: 10.1016/j.actbio.2015.04.018

[Display omitted] 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 90mg/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 90mg/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 570kPa and acellular scaffolds had a transparency of 87% in the 380–780nm 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%).