Clay/chitosan/poly(vinyl alcohol) nanofiber scaffold promotes differentiation of human dental pulp stem cells toward cholinergic and motor neuron subtypes.

This scientific research aimed to study the cholinergic and motor neuron-like cells differentiation potential of human dental pulp stem cells (hDPSCs) within a novel scaffold montmorillonite/poly (vinyl alcohol) (OMMT/PVA) nanofibrous mesh. The chitosan combined with different concentration of OMMT in PVA was synthesized and the main functional groups within OMMT/PVA nanofibers structure were then identified using ATR-FTIR. The probable cytotoxicity of the scaffolds was tested using MTT and LDH assays. hDPSCs underwent the differentiation process within the nanofibers with different concentrations of OMMT (1%, 3%, and 5%) in two separated phases. The differentiation capability of the cells was examined by analyzing the expression profiling of OCT-4, Nestin, GFAP, MBP, ChAT, MAP2, and Islet1, and Cresyl violet staining. Our findings proved that the scaffold could improve the viability and the proliferation rate of all experimental cell groups over the whole period of preinduction and induction phases. Moreover, the expression profile of genes indicated that the number of cells expressing neural-specific markers for those groups in the induction phase was substantially higher than that of the control group. The percentage of neural-specific markers positive cells increased in induced cells within 5% OMMT in PVA, which indicates that scaffolds provide hDPSCs with a suitable microenvironment for growing and differentiating toward desired neural cell lineages as well. Our finding shows that the OMMT/PVA fibrous scaffolds have a promising neuroprotective property and suitable potential for guiding of differentiation in the hDPSCs into motor neuron and cholinergic subtypes.