Selenium nanoparticles inclusion into chitosan hydrogels act as a chemical inducer for differentiation of PC12 cells into neuronal cells

Background and Objective: Biomaterials and nanomaterials have generated a great opportunity in regenerative medicine. Neurological disorders can result in permanent and severe derangement in motor and sensory functions. This study was conducted to examine the effects of selenium nanoparticles (Se NPs) as a chemical inducer for differentiation of PC12 cells into sympathetic-like neurons characterized by neurite outgrowth.Materials and Methods: Size, surface charge, the shape of Se NPs and the morphology of hydrogels were characterized by dynamic light scattering (DLS), zeta sizer, transmission electron microscopy (TEM) and scanning electron microscopy (SEM), respectively. DAPI staining, RT-PCR and western blot assays were used to evaluate cell attachment and mRNA and protein levels of neuronal markers, respectively.Results: The hydrodynamic size of Se NPs was about 33.55 nm and their surface charge was shifted from -24 to +3.4 mV. The morphological characterization demonstrated monodisperse spherical particles after coating with BSA. SEM images demonstrated that chitosan hydrogel containing Se NPs has suitable pore sizes for penetration of cells. DAPI staining and live/dead assay demonstrated the ability of cell attachment and biocompatibility of hydrogel, respectively. RT-PCR and western blot assays showed that neurite extension of differentiated PC12 cells can be linked to significantly increased mRNA levels of Map2, β-tubulin, increased protein levels of neurofilament-200 (NF200) as neuronal markers and decreased protein levels of ki67 protein as a proliferation marker.Conclusion: Collectively, our findings show that Se NPs can act as a chemical inducer for the differentiation of PC12 cells into sympathetic-like