Browsing by Author "Garagorri, Nerea"
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Item Acellular human corneal matrix sheets seeded with human adipose-derived mesenchymal stem cells integrate functionally in an experimental animal model(2015-03-01) Alio del Barrio, Jorge L.; Chiesa, Massimo; Garagorri, Nerea; Garcia-Urquia, Nerea; Fernandez-Delgado, Jorge; Bataille, Laurent; Rodriguez, Alejandra; Arnalich-Montiel, Francisco; Zarnowski, Tomasz; Álvarez de Toledo, Juan P.; Alio, Jorge L.; De Miguel, Maria P.; Tecnalia Research & Innovation; BiomaterialesPurpose: To evaluate the invivo 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 90μm thickness were decellularized, and their lack of cytotoxicity was assayed. The recellularization was achieved by the injection of 2×105 labeled h-ADASC in the graft followed by five days of cell culture. The grafts were implanted invivo 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 invitro 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.Item Biointegration of corneal macroporous membranes based on poly(ethyl acrylate) copolymers in an experimental animal model(2015-03-01) Del Barrio, Jorge L.Alió; Chiesa, Massimo; Ferrer, Gloria Gallego; Garagorri, Nerea; Briz, Nerea; Fernandez-Delgado, Jorge; Valls, Maria Sancho Tello; Botella, Carmen Carda; García-Tuñón, Ignacio; Bataille, Laurent; Rodriguez, Alejandra; Arnalich-Montiel, Francisco; Ribelles, Jose L.Gómez; Antolinos-Turpín, Carmen M.; Gómez-Tejedor, Jose A.; Alió, Jorge L.; De Miguel, Maria P.; Tecnalia Research & Innovation; SGCurrently available keratoprosthesis models (non-biological corneal substitutes) have a less than 75% graft survival rate at 2 years. We aimed at developing a model for keratoprosthesis based on the use of poly(ethyl acrylate) (PEA)-based copolymers, extracellular matrix-protein coating and colonization with adipose-derived mesenchymal stem cells. Human adipose tissue derived mesenchymal stem cells (h-ADASC) colonization efficiency of seven PEA-based copolymers in combination with four extracellular matrix coatings were evaluated in vitro. Then, macroporous membranes composed of the optimal PEA subtypes and coating proteins were implanted inside rabbit cornea. After a 3-month follow-up, the animals were euthanized, and the clinical and histological biointegration of the implanted material were assessed. h-ADASC adhered and survived when cultured in all PEA-based macroporous membranes. The addition of high hydrophilicity to PEA membranes decreased h-ADASC colonization in vitro. PEA-based copolymer containing 10% hydroxyethyl acrylate (PEA-HEA10) or 10% acrylic acid (PEA-AAc10) monomeric units showed the best cellular colonization rates. Collagen plus keratan sulfate-coated polymers demonstrated enhanced cellular colonization respect to fibronectin, collagen, or uncoated PEAs. In vivo implantation of membranes resulted in an extrusion rate of 72% for PEA, 50% for PEA-AAc10, but remarkably of 0% for PEA-HEA10. h-ADASC survival was demonstrated in all the membranes after 3 months follow-up. A slight reduction in the extrusion rate of h-ADASC colonized materials was observed. No significant differences between the groups with and without h-ADASC were detected respect to transparency or neovascularization. We propose PEA with low hydroxylation as a scaffold for the anchoring ring of future keratoprosthesis.Item Confocal and Histological Features After Poly(Ethylene Glycol) Diacrylate Corneal Inlay Implantation(2019-11) Bidaguren, Aritz; Mendicute, Javier; Madarieta, Iratxe; Garagorri, Nerea; Biomateriales; Tecnalia Research & InnovationPurpose: To evaluate the in vivo biocompatibility of photopolymerized poly(ethylene glycol) diacrylate (PEGDA) intrastromal inlays in rabbit corneas. Methods: Sixty-three eyes of 42 New Zealand rabbits were included. Manual intrastromal pockets were dissected in 42 eyes. PEGDA inlays were obtained using a specifically designed photomask and were inserted in the intrastromal pocket of 21 eyes (inlay group); the remaining 21 right eyes did not receive any implant (pocket-only group). Twenty-one eyes with no intervention were used as controls. In vivo confocal microscopy (IVCM) was performed at every visit. After 2 months, rabbits were sacrificed and corneas removed for histological analysis. Results: Corneas remained clear in all but two animals, and five cases of corneal neovascularization were seen (P = 0.2). Inlays remained stable without evidence of lateral or anterior migration, and no other complications were observed. No changes in anterior and posterior keratocyte density (P = 0.3 and P = 0.1, respectively) or endothelial cell density (P = 0.23) were observed between groups during the study time by IVCM. On pathology samples, thinning of the epithelium over the inlay area and epithelial hyperplasia over the edges were observed. A polygonal empty space with no evidence of PEGDA hydrogel within the midstroma was seen in the inlay group. Keratocytes were normal in shape and number in the vicinity of the PEGDA implant area. Conclusions: Photopolymerized PEGDA intrastromal inlays have shown relatively good safety and stability in rabbit corneas. Inlays were biostable in the corneal environment and remained transparent during follow up. Translational Relevance: The investigated PEGDA is promising for the development of biocompatible intrastromal implants.