Electrical stimulation of neural-differentiating iPSCs on novel coaxial electroconductive nanofibers

• Published in: Biomaterials science Vol. 9; no. 15; pp. 5359 - 5382
• Main Authors: Garrudo, Fábio F. F, Nogueira, Diogo E. S, Rodrigues, Carlos A. V, Ferreira, Flávio A, Paradiso, Patrizia, Colaço, Rogério, Marques, Ana C, Cabral, Joaquim M. S, Morgado, Jorge, Linhardt, Robert J, Ferreira, Frederico Castelo
• Format: Journal Article
• Language: English
• Published: Cambridge Royal Society of Chemistry 27.07.2021
• Subjects: Biodegradability; Biomarkers; Cells (biology); Differentiation (biology); Finite element method; Mechanical properties; Nanofibers; Neurological diseases; Neurons; Stimulation; Tissue engineering
• ISSN: 2047-4830; 2047-4849
• DOI: 10.1039/d1bm00503k

Neural tissue engineering strategies are paramount to create fully mature neurons, necessary for new therapeutic strategies for neurological diseases or the creation of reliable in vitro models. Scaffolds can provide physical support for these neurons and enable cues for enhancing neural cell differentiation, such as electrical current. Coaxial electrospinning fibers, designed to fulfill neural cell needs, bring together an electroconductive shell layer (PCL-PANI), able to mediate electrical stimulation of cells cultivated on fibers mesh surface, and a soft core layer (PGS), used to finetune fiber diameter (951 ± 465 nm) and mechanical properties (1.3 ± 0.2 MPa). Those dual functional coaxial fibers are electroconductive (0.063 ± 0.029 S cm −1 , stable over 21 days) and biodegradable (72% weigh loss in 12 hours upon human lipase accelerated assay). For the first time, the long-term effects of electrical stimulation on induced neural progenitor cells were studied using such fibers. The results show increase in neural maturation (upregulation of MAP2, NEF-H and SYP), up-regulation of glutamatergic marker genes (VGLUT1 - 15-fold) and voltage-sensitive channels (SCN1α - 12-fold, CACNA1C - 32-fold), and a down-regulation of GABAergic marker (GAD67 - 0.09-fold), as detected by qRT-PCR. Therefore, this study suggest a shift from an inhibitory to an excitatory neural cell profile. This work shows that the PGS/PCL-PANI coaxial fibers here developed have potential applications in neural tissue engineering. Coaxial PCL-PANI/PGS fibers are biodegradable and electroconductive (0.063 ± 0.029 S cm −1 ) for 21 days in PBS. Electrical stimulation of cultured iNPCs (30 days) lead to a dominant excitatory (VGLUT1) neural profile over an inhibitory (GAD67) one.