Controlling the fate of stem cells through two-and three-dimensional scaffolds based on bioresorbable polymers and graphenen derivatives: a study towards nerve tissue regeneration.

184 p.Neurological disorders are the major cause of long-term impairment and the second largest cause of death worldwide. Hence, there is an urgent need for new treatments that allow the functional recovery of damaged tissue. Among the experimental treatments, bioresorbable polyesters are showing great results in preclinical and clinical trials due to their biocompatibility, tunable degradability, versatility and physicochemical and mechanical properties. In this PhD thesis nanostructured scaffolds based on bioresorbable polymers and graphene oxide were developed to study the attachment, aligned growth and migration of both murine and human stem cells, avoiding the use of extracellular matrix-like compounds coatings. The use of murine neural stem cells allowed to study the differentiation pattern of the cells over the nanostructuredscaffolds, focusing on the achievement of a balanced neuronal and glial support, for a long-term survival of the cultures in vitro. The use of a relatively new source of stem cells, now considered clinical waste, like the dental pulp stem cells, allowed to minimize the ethical concerns and provide an actual alternative for personalized medicine in future therapies. To test this alternative, the regeneration capabilities of the nanostructured scaffolds were studied after the impairment of the rostral migratory stream in a rodent model in vivo. And with the aim of addressing the enhanced restoration capabilities of the personalized advanced medical products combining polymeric materials and human stem cells, the regeneration of the rostral migratory stream was compared when grafting the dental pulp stem cells, alone or in combination with our nanostructured scaffolds.Finally, to better resemble the neural niche in vitro graphene derivatives-based three-dimensional scaffolds with tunable geometry, mechanical and electrical conductive features were fabricated and their effect studied on cell survival and differentiation. Afterward, cerium oxide nanoparticles were incorporated to provide enhanced antioxidant and neuroprotective features and their effect on the establishment of balanced neuronal and glial co-cultures studied.Overall, this thesis gives new insights into the design of polymeric materials based on graphene derivatives for future personalized advanced medical products in combination with human stem cells for the restoration of the nervous syste

Lactide and Ethylene Brassylate-Based Thermoplastic Elastomers and Their Nanocomposites with Carbon Nanotubes: Synthesis, Mechanical Properties and Interaction with Astrocytes

Polylactide (PLA) is among the most commonly used polymers for biomedical applications thanks to its biodegradability and cytocompatibility. However, its inherent stiffness and brittleness are clearly inappropriate for the regeneration of soft tissues (e.g., neural tissue), which demands biomaterials with soft and elastomeric behavior capable of resembling the mechanical properties of the native tissue. In this work, both L- and D,L-lactide were copolymerized with ethylene brassylate, a macrolactone that represents a promising alternative to previously studied comonomers (e.g., caprolactone) due to its natural origin. The resulting copolymers showed an elastomeric behavior characterized by relatively low Young’s modulus, high elongation at break and high strain recovery capacity. The thermoplastic nature of the resulting copolymers allows the incorporation of nanofillers (i.e., carbon nanotubes) that further enable the modulation of their mechanical properties. Additionally, nanostructured scaffolds were easily fabricated through a thermo-pressing process with the aid of a commercially available silicon stamp, providing geometrical cues for the adhesion and elongation of cells representative of the nervous system (i.e., astrocytes). Accordingly, the lactide and ethylene brassylate-based copolymers synthesized herein represent an interesting formulation for the development of polymeric scaffolds intended to be used in the regeneration of soft tissues, thanks to their adjustable mechanical properties, thermoplastic nature and observed cytocompatibility.Grant PID2019-106236GB-I00 funded by MCIN/AEI/10.13039/501100011033. The authors are also thankful for funds from the Basque Government, Department of Education (IT-1766-22). C.B.-Á.: acknowledges the predoctoral grant funded by the UPV/EHU. Polimerbio and Y.P. have a Bikaintek Ph.D. grant (20-AF-W2-2018-00001)

Advances and Perspectives in Dental Pulp Stem Cell Based Neuroregeneration Therapies

Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types for regenerative therapies given their ability to grow in the absence of serum and their realistic possibility to be used in autologous grafts. In this review, we describe the particular advantages of hDPSCs for neuroregenerative cell therapies. We thoroughly discuss the knowledge about their embryonic origin and characteristics of their postnatal niche, as well as the current status of cell culture protocols to maximize their multilineage differentiation potential, highlighting some common issues when assessing neuronal differentiation fates of hDPSCs. We also review the recent progress on neuroprotective and immunomodulatory capacity of hDPSCs and their secreted extracellular vesicles, as well as their combination with scaffold materials to improve their functional integration on the injured central nervous system (CNS) and peripheral nervous system (PNS). Finally, we offer some perspectives on the current and possible future applications of hDPSCs in neuroregenerative cell therapies.This research was supported by MICINN retos I+D+i (PID2019-104766RB-C21 and RYC-2013-13450, to J.R.P.) and UPV/EHU (GIU16/66 and PPGA20/22, to F.U., G.I.; and COLAB19/03 and IKERTU-2020.0155, to F.U., J.R.S.). Y.P. was funded by a Bikaintek PhD grant from the Basque Government (20-AF-W2-2018-00001)