Differentiation of human dental pulp stem cells into cholinergic neurons using nanofibrous mesh

Introduction Cell transplantation is a viable strategy for treatment of neurodegenerative disorders. Combination of stem cells and nanobiomaterials for guiding neural lineage differentiation is expected to be a viable strategy in this regard. Objectives Nevertheless, providing a suitable cell source and an ideal bio-scaffold to support cell adhesion, migration and differentiation has remained challenging in neurodegenerative diseases cell therapy. MethodsIn this study, the dental pulp mesenchymal stem cells (DPSCs) were isolated from human third molar and characterized by flow cytometry and their differentiation capacity. The cells were seeded on a nanofibrous mesh made from 5% chitosan-montmorillonite/poly(vinyl alcohol) (CS-MMT/PVA) and then induced with neural lineage specific differentiation media. The cells cultured in cell culture plate served as negative control. The differentiation of the DPSCs before and after neural differentiation induction was studied by determining the expression level of stemness marker (Oct4: octamer-binding transcription factor 4), neuroprogenitor markers (nestin), cholinergic neurons (ChAT:choline o-acetyltransferase ) and mature neuronal markers (Map2: microtubule associated protein 2) using real-time PCR. The differentiation capacity of the DPSCs was further determined by immunostaining of nestin and ChAT.ResultsThe results indicated that the cell cultured on electrospun mesh significantly up-regulated the cholinergic markers of the cells after induction with neural lineage differentiation in comparison with negative control. ConclusionThe results strongly suggest the potential of DPSCs as an excellent cell source for neurodegenerative disorder cell transplantation and the CS-MMT/PVA as a promising supporter for DPSCs adhesion and differentiation toward cholinergic neuron

Keywords:

Dental pulp stem cells , Chitosan , Montmorillonite , Electrospinning , Cholinergic