Lactate-releasing PLA scaffolds for brain regeneration
Stroke and traumatic brain injuries are common causes of disability, with loss of nerve tissue due to secondary degeneration, gliosis, and often the formation of cavities that inhibit neural cell growth. Recent attempts at neural cell regeneration have therefore focused on the use of engineering mat...
| Autor: | |
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2014 |
| País: | España |
| Institución: | CBUC, CESCA |
| Repositorio: | TDR. Tesis Doctorales en Red |
| OAI Identifier: | oai:www.tdx.cat:10803/283142 |
| Acceso en línea: | http://hdl.handle.net/10803/283142 https://dx.doi.org/10.5821/dissertation-2117-95483 |
| Access Level: | acceso abierto |
| Palabra clave: | 576 616.8 620 |
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Lactate-releasing PLA scaffolds for brain regeneration |
| title |
Lactate-releasing PLA scaffolds for brain regeneration |
| spellingShingle |
Lactate-releasing PLA scaffolds for brain regeneration Álvarez Pinto, Zaida 576 616.8 620 |
| title_short |
Lactate-releasing PLA scaffolds for brain regeneration |
| title_full |
Lactate-releasing PLA scaffolds for brain regeneration |
| title_fullStr |
Lactate-releasing PLA scaffolds for brain regeneration |
| title_full_unstemmed |
Lactate-releasing PLA scaffolds for brain regeneration |
| title_sort |
Lactate-releasing PLA scaffolds for brain regeneration |
| dc.creator.none.fl_str_mv |
Álvarez Pinto, Zaida |
| author |
Álvarez Pinto, Zaida |
| author_facet |
Álvarez Pinto, Zaida |
| author_role |
author |
| dc.contributor.none.fl_str_mv |
Engel López, Elisabeth Alcántara Horrillo, Soledad Universitat Politècnica de Catalunya. Departament d'Enginyeria de Sistemes, Automàtica i Informàtica Industrial |
| dc.subject.none.fl_str_mv |
576 616.8 620 |
| topic |
576 616.8 620 |
| description |
Stroke and traumatic brain injuries are common causes of disability, with loss of nerve tissue due to secondary degeneration, gliosis, and often the formation of cavities that inhibit neural cell growth. Recent attempts at neural cell regeneration have therefore focused on the use of engineering materials that mimic the adult neural stem cell (NSC) niche, in order to establish an adequate environment for neurogenesis and differentiation. However, because the adult mammalian NSC niche has limited regenerative capacities, effective regeneration of the central nervous system (CNS) requires the reconstitution of its embryonic counterpart. Radial glia are bipolar cells with 1-2 µm-thick shafts that form a palisade and span the entire CNS parenchyma serving as substrates for neuronal migration. They contain high levels of glycogen and release L-lactate. Cerebral energy metabolism is a highly compartmentalized and complex process. The adult brain normally uses glucose as its primary energy source. However, before and immediately after birth, lactate is also an important energy source because at this time the level of glucose is low. Over the past decade, a role for lactate in fuelling the energetic requirements of neurons has emerged, not only during the perinatal period but also in adulthood. Initial evidence suggests that the metabolisms of NSC, neurons and astrocytes differ and that energy-dependent processes may influence the balance between NSC self-renewal and differentiation. The main goal of this thesis was to design an implantable biomaterial scaffold that reproduces the organization and supportive function of embryonic radial glia. Here we tested two types of poly L/DL lactic acid (PLA95/5 and PLA70/30), a biodegradable material permissive for neural cell adhesion and growth, as materials for nerve regeneration. PLA95/5 films were highly crystalline, stiff (GPa), and did not degrade significantly in the period analyzed in culture. In contrast, PLA70/30 films were more amorphous, softer (MPa) and degraded faster, releasing significant amounts of lactate into the medium. PLA70/30 performed better than PLA95/5 for primary cortical neural cell adhesion, proliferation and differentiation, maintaining the pools of neuronal and glial progenitor cells in vitro. Finally, for in vivo studies, we designed 3D cell-free biomimetic scaffolds consisting of electrospun PLA70/30 nanofibers. Radially aligned scaffolds released L-lactate and reproduced the 3D organization and supportive function of radial glia. These scaffolds implanted into cavities made in mouse brain fostered complete implant vascularization, sustained neurogenesis, and allowed the long-term survival and integration of the newly generated neurons. Our results suggest that PLA70/30 scaffolds mimic some of the physical and biochemical characteristics of the NSC niche. Overall, our results show that the endogenous CNS is capable of regeneration through the in vivo dedifferentiation induced by biophysical and metabolic cues, with no need for exogenous cells, growth factors, or genetic manipulation. |
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2014 |
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2014 2014 2014 |
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info:eu-repo/semantics/doctoralThesis info:eu-repo/semantics/publishedVersion |
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doctoralThesis |
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publishedVersion |
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http://hdl.handle.net/10803/283142 https://dx.doi.org/10.5821/dissertation-2117-95483 |
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http://hdl.handle.net/10803/283142 https://dx.doi.org/10.5821/dissertation-2117-95483 |
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Inglés |
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Inglés |
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http://creativecommons.org/licenses/by-nc-nd/3.0/es/ info:eu-repo/semantics/openAccess |
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http://creativecommons.org/licenses/by-nc-nd/3.0/es/ |
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openAccess |
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330 p. application/pdf application/pdf |
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Universitat Politècnica de Catalunya |
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Universitat Politècnica de Catalunya |
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TDX (Tesis Doctorals en Xarxa) reponame:TDR. Tesis Doctorales en Red instname:CBUC, CESCA |
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CBUC, CESCA |
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TDR. Tesis Doctorales en Red |
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TDR. Tesis Doctorales en Red |
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1869419578515259392 |
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Lactate-releasing PLA scaffolds for brain regenerationÁlvarez Pinto, Zaida576616.8620Stroke and traumatic brain injuries are common causes of disability, with loss of nerve tissue due to secondary degeneration, gliosis, and often the formation of cavities that inhibit neural cell growth. Recent attempts at neural cell regeneration have therefore focused on the use of engineering materials that mimic the adult neural stem cell (NSC) niche, in order to establish an adequate environment for neurogenesis and differentiation. However, because the adult mammalian NSC niche has limited regenerative capacities, effective regeneration of the central nervous system (CNS) requires the reconstitution of its embryonic counterpart. Radial glia are bipolar cells with 1-2 µm-thick shafts that form a palisade and span the entire CNS parenchyma serving as substrates for neuronal migration. They contain high levels of glycogen and release L-lactate. Cerebral energy metabolism is a highly compartmentalized and complex process. The adult brain normally uses glucose as its primary energy source. However, before and immediately after birth, lactate is also an important energy source because at this time the level of glucose is low. Over the past decade, a role for lactate in fuelling the energetic requirements of neurons has emerged, not only during the perinatal period but also in adulthood. Initial evidence suggests that the metabolisms of NSC, neurons and astrocytes differ and that energy-dependent processes may influence the balance between NSC self-renewal and differentiation. The main goal of this thesis was to design an implantable biomaterial scaffold that reproduces the organization and supportive function of embryonic radial glia. Here we tested two types of poly L/DL lactic acid (PLA95/5 and PLA70/30), a biodegradable material permissive for neural cell adhesion and growth, as materials for nerve regeneration. PLA95/5 films were highly crystalline, stiff (GPa), and did not degrade significantly in the period analyzed in culture. In contrast, PLA70/30 films were more amorphous, softer (MPa) and degraded faster, releasing significant amounts of lactate into the medium. PLA70/30 performed better than PLA95/5 for primary cortical neural cell adhesion, proliferation and differentiation, maintaining the pools of neuronal and glial progenitor cells in vitro. Finally, for in vivo studies, we designed 3D cell-free biomimetic scaffolds consisting of electrospun PLA70/30 nanofibers. Radially aligned scaffolds released L-lactate and reproduced the 3D organization and supportive function of radial glia. These scaffolds implanted into cavities made in mouse brain fostered complete implant vascularization, sustained neurogenesis, and allowed the long-term survival and integration of the newly generated neurons. Our results suggest that PLA70/30 scaffolds mimic some of the physical and biochemical characteristics of the NSC niche. Overall, our results show that the endogenous CNS is capable of regeneration through the in vivo dedifferentiation induced by biophysical and metabolic cues, with no need for exogenous cells, growth factors, or genetic manipulation.Las lesiones cerebrales son causas comunes de discapacidad que conllevan pérdida de tejido nervioso debido a la degeneración secundaria, la gliosis, y con frecuencia la formación de cavidades que inhiben el crecimiento neuronal. Recientemente, las terapias neuroregenerativas se han centrado en el uso de la ingeniería de materiales. Durante el desarrollo del SNC, las células de glia radial son las principales CMN que generan neuronas y glía y son retenidas en el cerebro adulto de especies que regeneran. Durante la neurogénesis temprana, los vasos sanguíneos invaden el SNC e interactúan con las CMN, dando lugar al nicho neurovascular. En el cerebro adulto, la glia radial puede ser recuperada al menos en cierta medida después de una lesión, lo que indica un intento endógeno en la reconstitución del nicho embrionario. Las células de glía radial son bipolares con 1-2 micras de espesor que forman una empalizada que abarca todo el parénquima del SNC sirviendo como sustrato para la migración neuronal. Esta glía radial contiene altos niveles de glucógeno liberando al exterior L-lactato. El metabolismo energético cerebral es un proceso altamente compartimentado y complejo. El cerebro adulto normalmente usa glucosa como fuente de energía primaria. Sin embargo, antes e inmediatamente después del nacimiento, el lactato es también una fuente de energía importante, debido a que los niveles de glucosa son bajos. En la última década, se ha visto que el lactato juega un papel importante como factor energético para las neuronas, no sólo durante el período perinatal sino también en la edad adulta. Nuevas evidencias sugieren que el metabolismo de las CMN, neuronas y astrocitos son diferentes y que los procesos dependientes de energía pueden influir en el equilibrio entre la auto-renovación y diferenciación de las CMN. El objetivo principal de esta tesis ha sido diseñar un andamio implantable que reprodujera la organización y función de soporte de la glía radial embrionaria. Para ello, hemos probado dos tipos de ácido poli L/DL láctico (PLA95/5 y PLA70/30), material biodegradable y permisivo para la adhesión y el crecimiento neuronal. Las películas de PLA95/5 eran cristalinas, rígidas (GPa), y no se degradaban significativamente en el período analizado durante el cultivo in vitro. Sin embargo, las películas de PLA70/30 eran más amorfas, menos rígidas (MPa) y se degradaban más rápido, liberando cantidades significativas de lactato en el medio. A diferencia del PLA95/5, en el 70/30 había una mejor adhesión neuronal, así como la proliferación y el mantenimiento de los progenitores neuronales y gliales in vitro. Finalmente, para los estudios in vivo, diseñamos andamios biomiméticos en 3D libres de células que consistían en nanofibras de PLA70/30. Los andamios radialmente alineados reproducían la organización 3D y la función de apoyo de la glía radial. Estos andamios implantados en la corteza cerebral del ratón ayudaron a la completa vascularización del implante, permitiendo la supervivencia y la integración a largo plazo de las neuronas recién generadas. Nuestros resultados sugieren que los andamios de PLA70/30 imitan algunas de las características físicas y bioquímicas del nicho neurovascular. En conclusión, nuestros resultados muestran que el SNC es capaz de regenerar de manera endógena a través de la des-diferenciación in vivo inducida por las señales biofísicas y metabólicas, sin necesidad de células exógenas, factores de crecimiento o manipulación genéticaDOCTORAT EN ENGINYERIA BIOMÈDICA (Pla 2007)Universitat Politècnica de CatalunyaEngel López, ElisabethAlcántara Horrillo, SoledadUniversitat Politècnica de Catalunya. Departament d'Enginyeria de Sistemes, Automàtica i Informàtica Industrial201420142014info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersion330 p.application/pdfapplication/pdfhttp://hdl.handle.net/10803/283142https://dx.doi.org/10.5821/dissertation-2117-95483TDX (Tesis Doctorals en Xarxa)reponame:TDR. Tesis Doctorales en Redinstname:CBUC, CESCAInglésL'accés als continguts d'aquesta tesi queda condicionat a l'acceptació de les condicions d'ús establertes per la següent llicència Creative Commons: http://creativecommons.org/licenses/by-nc-nd/3.0/es/http://creativecommons.org/licenses/by-nc-nd/3.0/es/info:eu-repo/semantics/openAccessoai:www.tdx.cat:10803/2831422026-06-14T12:46:07Z |
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15.300719 |