title: Acellular human corneal matrix sheets seeded with human adipose-derived mesenchymal stem cells integrate functionally in an experimental animal model.
abstract: OBJECTIVE
  To evaluate the in vivo biocompatibility of grafts composed of sheets of decellularized human corneal stroma with or without the recellularization of human adipose derived adult stem cells (h-ADASC) into the rabbit cornea.
  
  
  METHODS
  Sheets of human corneal stroma of 90 μm thickness were decellularized, and their lack of cytotoxicity was assayed. The recellularization was achieved by the injection of 2 × 10(5) labeled h-ADASC in the graft followed by five days of cell culture. The grafts were implanted in vivo into a stromal pocket at 50% depth. After a triple-masked three-month follow-up, the animals were euthanized and the biointegration of the graft, the viability of the stem cells and the expression of keratocan (human keratocyte-specific protein) were assessed.
  
  
  RESULTS
  The decellularized stromal sheets showed an intact extracellular matrix with a decellularization rate of 92.8% and an excellent recellularization capacity in vitro with h-ADASC. A complete and stable graft transparency was observed during the full follow-up, with absence of any clinical sign of rejection. The postmortem analysis demonstrated the survival of the transplanted human stem cells inside the graft and their differentiation into functional keratocytes, as assessed by the expression of human keratocan.
  
  
  CONCLUSIONS
  We report a new model of lamellar keratoplasty that requires only a simple and safe procedure of liposuction and a donor allogeneic cornea to provide an optically transparent autologous stromal graft with excellent biocompatibility and integration into the host tissue in a rabbit model.
authors.affiliation: Ophthalmology Department, Ramon y Cajal Hospital, Madrid, Spain.
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Alio del Barrio
authors.affiliation: Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain.
authors.forename: Massimo
authors.initials: M
authors.lastname: Chiesa
authors.affiliation: Tecnalia Research & Innovation, Health Division-Biomaterials Area, Mikeletegi Pasealekua 2, 20009 Donostia-San Sebastian, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: Garagorri
authors.affiliation: Tecnalia Research & Innovation, Health Division-Biomaterials Area, Mikeletegi Pasealekua 2, 20009 Donostia-San Sebastian, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: Garcia-Urquia
authors.affiliation: Department of Plastic and Reconstructive Surgery, Santa Cristina Hospital and Centrocim, Madrid, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Fernandez-Delgado
authors.affiliation: Vissum Ophthalmological Institute and Miguel Hernandez University, Alicante, Spain.
authors.forename: Laurent
authors.initials: L
authors.lastname: Bataille
authors.affiliation: Vissum Ophthalmological Institute and Miguel Hernandez University, Alicante, Spain.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Rodriguez
authors.affiliation: Ophthalmology Department, Ramon y Cajal Hospital, Madrid, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Arnalich-Montiel
authors.affiliation: Lublin Eye Bank, Lublin, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Zarnowski
authors.affiliation: Eye Bank of Barraquer Ophthalmology Center, Barcelona, Spain.
authors.forename: Juan P
authors.initials: JP
authors.lastname: Álvarez de Toledo
authors.affiliation: Vissum Ophthalmological Institute and Miguel Hernandez University, Alicante, Spain.
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Alio
authors.affiliation: Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain. Electronic address: mariapdemiguel@gmail.com.
authors.forename: Maria P
authors.initials: MP
authors.lastname: De Miguel
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D003316:Corneal Diseases / Q000601:surgery*; D003319:Corneal Stroma / Q000166:cytology / Q000637:transplantation*; D016039:Corneal Transplantation / Q000379:methods*; D004195:Disease Models, Animal; D005109:Extracellular Matrix; D006801:Humans; D059630:Mesenchymal Stem Cells; D011817:Rabbits; D033581:Stem Cell Transplantation; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds; D014184:Transplantation, Homologous
keywords: Adipose derived stem cells; Cornea; Corneal stroma regeneration; Decellularization; Lamellar corneal transplant; Recellularization; Tissue engineering