Electrospun conductive nanofiber yarns for accelerating mesenchymal stem cells differentiation and maturation into Schwann cell-like cells under a combination of electrical stimulation and chemical induction

Abstract

Development of multifunctional tube-filling materials is required to improve the performances of the existing nerve guidance conduits (NGCs) in the repair of long-gap peripheral nerve (PN) injuries. In this study, composite nanofiber yarns (NYs) based on poly(p-dioxanone) (PPDO) biopolymer and different concentrations of carbon nanotubes (CNTs) were manufactured by utilizing a modified electrospinning apparatus. We confirmed the successful incorporation of CNTs into the PPDO nanofibers of as-fabricated composite NYs. The PPDO/CNT NYs exhibited similar morphology and structure in comparison with pure PPDO NYs. However, the PPDO/CNT NYs showed obviously enhanced mechanical properties and electrical conductivity compared to PPDO NYs. The biological tests revealed that the addition of CNTs had no negative effects on the cell growth, and proliferation of rabbit Schwann cells (rSCs), but it better maintained the phenotype of rSCs. We also demonstrated that the electrical stimulation (ES) significantly enhanced the differentiation capability of human adipose-derived mesenchymal stem cells (hADMSCs) into SC-like cells (SCLCs) on the PPDO/CNT NYs. More importantly, a unique combination of ES and chemical induction was found to further enhance the maturation of hADMSC-SCLCs on the PPDO/CNT NYs by notably upregulating the expression levels of SC myelination-associated gene markers and increasing the growth factor secretion. Overall, this study showed that our electrically conductive PPDO/CNT composite NYs could provide a beneficial microenvironment for various cell activities, making them an attractive candidate as NGC-infilling substrates for PN regeneration applications. STATEMENT OF SIGNIFICANCE: The morphology, microstructure, and bioelectrical properties of conductive PPDO/CNT NYs have been explored for guiding or controlling cell behaviors. The PPDO/CNT NYs exhibited improved mechanical properties and increased electrical conductivity compared to the CNT-free PPDO NYs. They also presented an obviously enhanced biocompatibility by effectively maintaining the phenotype of rSCs. In addition, when hADMSCs were seeded and cultured on the conductive PPDO/CNT NYs, CI was demonstrated to promote the SC-related growth factor secretion of hADMSCs, and ES was demonstrated to improve the phenotypic maturation of hADMSCs into myelinating SCLCs. Moreover, the combination of CI and ES was found to further synergistically enhance the maturation of hADMSC-SCLCs. The achievement of conductive PPDO/CNT NYs shows potential for application as NGC-infilling substrates for PN regeneration.

Keywords: Cell differentiation; Electrical stimulation; Electrospinning; Myelination; Nerve regeneration.