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dc.creatorSilva J-
dc.creatorBento AR-
dc.creatorBarros D-
dc.creatorLaundos TL-
dc.creatorSousa SR-
dc.creatorQuelhas P-
dc.creatorSousa MM-
dc.creatorPêgo AP-
dc.creatorAmaral IF-
dc.description.abstractTo enhance fibrin hydrogel affinity towards pluripotent stem cell-derived neural stem/progenitor cells (NSPCs) and its capacity to support NSPC migration and neurite extension, we explored the tethering of synthetic peptides engaging integrin a6ß1, a cell receptor enriched in NSPCs. Six a6ß1 integrin ligands were tested for their ability to support integrin a6ß1-mediated adhesion of embryonic stem cell-derived NSPCs (ES-NSPs) and sustain ES-NSPC viability, migration, and neuronal differentiation. Due to their better performance, peptides T1, HYD1, and A5G81 were immobilized into fibrin and functionalized gels characterized in terms of peptide binding efficiency, structure and viscoelastic properties. Tethering of T1 or HYD1 successfully enhanced cell outgrowth from ES-NSPC neurospheres (up to 2.4-fold increase), which exhibited a biphasic response to peptide concentration. Inhibition assays evidenced the involvement of a6ß1 and a3ß1 integrins in mediating radial outgrowth on T1-/HYD1-functionalized gels. Fibrin functionalization also promoted neurite extension of single ES-NSPCs in fibrin, without affecting cell proliferation and neuronal differentiation. Finally, HYD1-functionalized gels were found to provide a permissive environment for axonal regeneration, leading up to a 2.0-fold increase in neurite extension from rat dorsal root ganglia explants as compared to unmodified fibrin, and to significant improved locomotor function after spinal cord injury (complete transection), along with a trend toward a higher area positive for growth associated protein 43 (marker for axonal growth cone formation). Our results suggest that conjugation of a6ß1 integrin-binding motifs is of interest to increase the biofunctionality of hydrogels used in 3D platforms for ES-NSPC culture and potentially, in matrix-assisted ES-NSPC transplantation. Statement of Significance Impact statement: The transplantation of NSPCs derived from pluripotent stem cells holds much promise for the treatment of central nervous system disorders. Moreover, the combinatorial use of biodegradable hydrogels with NSPCs was shown to contribute to the establishment of a more permissive environment for survival and integration of transplanted cells. In this study, fibrin hydrogels functionalized with a synthetic peptide engaging integrin a6ß1 (HYD1) were shown to promote neurite extension of ES-NSPCs, which is fundamental for the formation of functional neuronal relay circuits after NSPC transplantation. Notably, HYD1-functionalized fibrin per se led to enhanced axonal growth ex vivo and to an improvement in locomotor function after implantation in a rat model of spinal cord injury. Conjugation of a6ß1 integrin-binding motifs may therefore be of interest to confer bioactivity to NSPC hydrogel vehicles.-
dc.description.sponsorshipThe authors would like to acknowledge Prof. Domingos Henrique (Instituto de Medicina Molecular, Lisbon) for providing theES 46C cell line. This work was supported by FEDER funds throughthe Programa Operacional Factores de Competitividade – COMPETE(FCOMP-01-0124-FEDER-021125) and by National Funds throughFCT – Fundação para a Ciência e a Tecnologia (PTDC/SAU-BMA/118869/2010). A.R.B. and D.B. are supported by FCT (SFRH/BD/86200/2012; PD/BD/105953/2014).-
dc.relation.ispartofActa Biomaterialia, vol.59, p. 243-256-
dc.subject.meshCell Line, Tumor-
dc.subject.meshEmbryonic Stem Cells-
dc.subject.meshIntegrin alpha6beta1-
dc.subject.meshNeural Stem Cells-
dc.subject.meshRats, Wistar-
dc.titleFibrin functionalization with synthetic adhesive ligands interacting with a6ß1 integrin receptor enhance neurite outgrowth of embryonic stem cell-derived neural stem/progenitors-
dc.typeArtigo em Revista Científica Internacional-
dc.contributor.uportoInstituto de Investigação e Inovação em Saúde-
Appears in Collections:I3S - Artigo em Revista Científica Internacional

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