Electrical Stimulation of Human Mesenchymal Stem Cells on Conductive Chitosan-Polyaniline Substrates Promotes Neural Priming

Abstract Electrical stimulation (ES) within conductive polymer substrates has been suggested to promote the differentiation of stem cells toward a neuronal phenotype. The use of conductive scaffolds in tissue regeneration provides a unique and attractive new option to control the amount and location of ES delivery. Scaffold stiffness has also been shown to be an important regulator of stem cells’ behavior and fate. Therefore, to improve stem cell-based regenerative therapies, it is essential to characterize the simultaneous effects of electroconductive substrate stiffness and electric field stimuli on stem cell fate processes. In this study, biodegradable electroconductive substrates based on chitosan-polyaniline (CS-g-PANI) were fabricated with different stiffnesses. Human mesenchymal stem cells (hMSCs) seeded on these scaffolds were electrically stimulated for 14 days with 100 mV/ cm (20 min every day). For hMSCs cultured on soft conductive scaffolds, a morphological change with significant filopodial elongation was observed after 2 weeks of electrically stimulated culture. Compared with stiff conductive CS-g-PANI scaffolds and non-conductive CS scaffolds, for soft conductive CS-g-PANI scaffolds microtubule-associated protein 2 (MAP2) and neurofilament (NF-H) expression increased after application of ES. At the same time, there was a decrease in the expression of the glial markers glial fibrillary acidic protein (GFAP) and vimentin after ES. Furthermore, the elevation of intracellular calcium [Ca 2+ ] during spontaneous, cell-generated Ca 2+ transients further suggested that electric field stimulation of hMSCs cultured on conductive CS-g-PANI substrates can promote a neural-like phenotype. Our findings propose that the combination of the soft conductive CS-g-PANI substrate and ES is a promising new tool for enhancing nerve tissue engineering outcomes.