title: Human Bone Derived Collagen for the Development of an Artificial Corneal Endothelial Graft. In Vivo Results in a Rabbit Model.
abstract: Corneal keratoplasty (penetrating or lamellar) using cadaveric human tissue, is nowadays the main treatment for corneal endotelial dysfunctions. However, there is a worldwide shortage of donor corneas available for transplantation and about 53% of the world's population have no access to corneal transplantation. Generating a complete cornea by tissue engineering is still a tough goal, but an endothelial lamellar graft might be an easier task. In this study, we developed a tissue engineered corneal endothelium by culturing human corneal endothelial cells on a human purified type I collagen membrane. Human corneal endothelial cells were cultured from corneal rims after corneal penetrating keratoplasty and type I collagen was isolated from remnant cancellous bone chips. Isolated type I collagen was analyzed by western blot, liquid chromatography -mass spectrometry and quantified using the exponentially modified protein abundance index. Later on, collagen solution was casted at room temperature obtaining an optically transparent and mechanically manageable membrane that supports the growth of human and rabbit corneal endothelial cells which expressed characteristic markers of corneal endothelium: zonula ocluddens-1 and Na+/K+ ATPase. To evaluate the therapeutic efficiency of our artificial endothelial grafts, human purified type I collagen membranes cultured with rabbit corneal endothelial cells were transplanted in New Zealand white rabbits that were kept under a minimal immunosuppression regimen. Transplanted corneas maintained transparency for as long as 6 weeks without obvious edema or immune rejection and maintaining the same endothelial markers that in a healthy cornea. In conclusion, it is possible to develop an artificial human corneal endothelial graft using remnant tissues that are not employed in transplant procedures. This artificial endothelial graft can restore the integrality of corneal endothelium in an experimental model of endothelial dysfunction. This strategy could supply extra endothelial tissue and compensate the deficit of cadaveric grafts for corneal endothelial transplantation.
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Natalia
authors.initials: N
authors.lastname: Vázquez
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Manuel
authors.initials: M
authors.lastname: Chacón
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Rodríguez-Barrientos
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Jesús
authors.initials: J
authors.lastname: Merayo-Lloves
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Miguel
authors.initials: M
authors.lastname: Naveiras
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).|Hospital Universitario Central de Asturias (Spain).
authors.forename: Begoña
authors.initials: B
authors.lastname: Baamonde
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Jose F
authors.initials: JF
authors.lastname: Alfonso
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Iriana
authors.initials: I
authors.lastname: Zambrano-Andazol
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).
authors.forename: Ana C
authors.initials: AC
authors.lastname: Riestra
authors.affiliation: Instituto Universitario Fernández-Vega. Universidad de Oviedo (Spain).|CIBER on rare disease-CIBERer. Instituto de Investigaciones Sanitarias de la Fundación Jiménez Díaz-IIS-FJD (Spain).
authors.forename: Álvaro
authors.initials: Á
authors.lastname: Meana
mesh_terms: D000818:Animals; D024042:Collagen Type I / Q000378:metabolism / Q000627:therapeutic use; D003315:Cornea / Q000473:pathology; D003317:Corneal Dystrophies, Hereditary / Q000503:physiopathology / Q000628:therapy*; D016039:Corneal Transplantation; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D006801:Humans; D015948:Keratoplasty, Penetrating / Q000379:methods; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D023822:Tissue Engineering; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D062826:Zonula Occludens-1 Protein; D000254:Sodium-Potassium-Exchanging ATPase