Evaluation of Small Intestine Grafts Decellularization Methods for Corneal Tissue Engineering
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AutorXimenes Oliveira, Ana Celeste; Garzón Bello, Ingrid Johanna; Ionescu, Ana Maria; Carriel Araya, Víctor; Cruz Cardona, Juan de la; González Andrades, Miguel; Pérez, María del Mar; Alaminos Mingorance, Miguel; Campos Muñoz, Antonio
Public Library of Science (PLOS)
Extracellular-matrixStroma substituteScaffold materialCollagen-fibersCross-linkingSubmucosaFibrinOrganReconstructionRegeneration
Oliveira, A.C.; et al. Evaluation of Small Intestine Grafts Decellularization Methods for Corneal Tissue Engineering. Plos One, 8(6): e66538 (2013). [http://hdl.handle.net/10481/30758]
PatrocinadorThis study was supported by the Spanish Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica (I+D+I), Instituto de Salud Carlos III (ISCIII), grant FIS PI11/2680, and by the Regional Ministry of Health, grant SAS PI-0462-2010, Junta de Andalucía Spain. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Advances in the development of cornea substitutes by tissue engineering techniques have focused on the use of decellularized tissue scaffolds. In this work, we evaluated different chemical and physical decellularization methods on small intestine tissues to determine the most appropriate decellularization protocols for corneal applications. Our results revealed that the most efficient decellularization agents were the SDS and triton X-100 detergents, which were able to efficiently remove most cell nuclei and residual DNA. Histological and histochemical analyses revealed that collagen fibers were preserved upon decellularization with triton X-100, NaCl and sonication, whereas reticular fibers were properly preserved by decellularization with UV exposure. Extracellular matrix glycoproteins were preserved after decellularization with SDS, triton X-100 and sonication, whereas proteoglycans were not affected by any of the decellularization protocols. Tissue transparency was significantly higher than control non-decellularized tissues for all protocols, although the best light transmittance results were found in tissues decellularized with SDS and triton X-100. In conclusion, our results suggest that decellularized intestinal grafts could be used as biological scaffolds for cornea tissue engineering. Decellularization with triton X-100 was able to efficiently remove all cells from the tissues while preserving tissue structure and most fibrillar and non-fibrillar extracellular matrix components, suggesting that this specific decellularization agent could be safely used for efficient decellularization of SI tissues for cornea TE applications.