Gelatinan oinarritutako hiru dimentsiotako sistema egokituak hezur ehunen ingeniaritzarako
28 visitas recibidas
Autoría:
Maria del Carmen Echave Otaño
Director/a:
Gorka Orive Arroyo, José Luis Pedraz Muñoz
Universidad:
Universidad del País Vasco
Facultad:
Facultad de Farmacia
Departamento:
Farmacia y Ciencias de los Alimentos
Área:
Medicina
Año:
2019
Idioma:
Inglés
|Doktorego-tesi honen helburua da hiru dimentsioko sistemak garatzea hezur-ehunen ingeniaritzarako.Gelatina aukeratu da material nagusi gisa, eta harekin garatu dira hazkunde-faktoreen eramaile eta terapia zelularrerako euskarri gisa baliatzeko aldamio-egiturak.Sarreran, jatorri naturaleko polimero horren ezaugarri fisiko-kimiko eta biologiko nagusiak aurkezten dira, bai eta hezur-ehunaren ingeniaritzarako gelatinan oinarritutako ikuspegietan berriki izandako aurrerapenak ere.Lehen lan esperimentalean, gelatinaren entzima-erretikulazioa optimizatu da mikrobio-transglutaminasa entzimarekin, eta VEGF faktorearen eta BMP-2 proteinaren in vitro askapenaren ezaugarriak zehaztu dira.Zelula-kultiboaren ondoren, formulazio horien errendimendu biologikoa ebaluatu zen.Hurrengo azterketan, hidroxiapatitarekin eta kaltzio-sulfatoarekin indartu genituen gelatinaz eginiko aldamio-egiturok.In vitro karakterizazioa eta hezurrak in vivo birsortzeko gaitasuna ebaluatu ziren, sagu osteoporotikoen eredua baliatuz.BMP-2 proteinaz aberastutako gelatina-sistema indartuek arrakastaz konpondu zituzten hezurretako akats kritikoak.Azkenik, gelatina mineralizatuz osatutako konposatu bat eta gelatina anisotropiko bioinspiratuzko hidrogel bat integratu genituen hiru dimentsioko sistema gradual bakarrean, interfaze muskuloeskeletikoetan dauden konposizio, arkitektura eta antolamendu zelularrak emulatzeko.In vitro ebaluazio biologikoa, berriz, hiru dimentsioko hidrogel bifasikoaren barruan kapsulatutako gantz-ehunetik eratorritako giza zelula amekin egin zen.The current Doctoral Thesis is focused on the development of three-dimensional systems for bone tissue engineering purposes. Gelatin was chosen as the main material to manufacture scaffolds capable for serving as both growth factors delivery carrier and cell-therapy support. The main physic-chemical and biological properties of this natural origin polymer, together with the recent advances in gelatin-based approaches for bone tissue engineering are presented in the Introduction part. In the first experimental work, enzymatic crosslinking of gelatin with microbial transglutaminase enzyme has been optimized and in vitro release of VEGF and BMP-2 were characterized. Biological performance of those formulations was evaluated after cell seeding. In the following study, we reinforced the gelatin-based scaffolds with hydroxyapatite and calcium sulfate. In vitro characterization and in vivo bone regeneration capacity was assessed with osteoporotic mice animal model. Reinforced gelatin systems loaded with BMP-2 promoted successful bone repair in the critical sized defects. Finally, we integrated mineralized gelatin composite and bioinspired anisotropic gelatin hydrogel in a single gradual 3D system, in order to emulate the dissimilar composition, architecture and cellular organization found in the musculoskeletal interfaces. The in vitro biological assessment was performed with human adipose-derived stem cell encapsulated within the 3D biphasic hydrogel.
