Jorge L Alio del Barrio1, Massimo Chiesa2, Nerea Garagorri3, Nerea Garcia-Urquia3, Jorge Fernandez-Delgado4, Laurent Bataille5, Alejandra Rodriguez5, Francisco Arnalich-Montiel1, Tomasz Zarnowski6, Juan P Álvarez de Toledo7, Jorge L Alio5, Maria P De Miguel8. 1. Ophthalmology Department, Ramon y Cajal Hospital, Madrid, Spain. 2. Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain. 3. Tecnalia Research & Innovation, Health Division-Biomaterials Area, Mikeletegi Pasealekua 2, 20009 Donostia-San Sebastian, Spain. 4. Department of Plastic and Reconstructive Surgery, Santa Cristina Hospital and Centrocim, Madrid, Spain. 5. Vissum Ophthalmological Institute and Miguel Hernandez University, Alicante, Spain. 6. Lublin Eye Bank, Lublin, Poland. 7. Eye Bank of Barraquer Ophthalmology Center, Barcelona, Spain. 8. Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain. Electronic address: mariapdemiguel@gmail.com.
Abstract
PURPOSE: 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.
PURPOSE: To evaluate the in vivo biocompatibility of grafts composed of sheets of decellularized humancorneal stroma with or without the recellularization of human adipose derived adult stem cells (h-ADASC) into the rabbit cornea. METHODS: Sheets of humancorneal 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: Marcus Ang; Damien Gatinel; Dan Z Reinstein; Erik Mertens; Jorge L Alió Del Barrio; Jorge L Alió Journal: Eye (Lond) Date: 2020-07-24 Impact factor: 3.775
Authors: Christopher J Rogers; Robert J Harman; Bruce A Bunnell; Martin A Schreiber; Charlie Xiang; Fu-Sheng Wang; Antonio F Santidrian; Boris R Minev Journal: J Transl Med Date: 2020-05-18 Impact factor: 5.531