Magnetic chitosan hydrogel induces neuronal differentiation of neural stem cells by activating RAS-dependent signal cascade

Our aim was to modulate magnetic cues to influence the differentiation of neural stem cell (NSC) into neuron during nerve repair and to explore corresponding mechanisms. Here, a magnetic hydrogel composed of chitosan matrices and magnetic nanoparticles (MNPs) with different content was prepared as the magnetic-stimulation platform to apply intrinsically-present magnetic cue and externally-applied magnetic field to NSC grown on the hydrogel. The MNP content had regulatory effects on neuronal differentiation and the MNPs-50 samples exhibited the best neuronal potential and appropriate biocompatibility in vitro, as well as accelerated the subsequent neuronal regeneration in vivo. Remarkably, the use of proteomics analysis parsed the underlying mechanism of magnetic cue-mediated neuronal differentiation form the perspective of protein corona and intracellular signal transduction. The intrinsically-present magnetic cues in hydrogel contributed to the activation of intracellular RAS-dependent signal cascades, thus facilitating neuronal differentiation. Magnetic cue-dependent changes in NSCs benefited from the upregulation of adsorbed proteins related to "neuronal differentiation", "cell-cell interaction", "receptor", "protein activation cascade", and "protein kinase activity" in the protein corona. Additionally, magnetic hydrogel acted cooperatively with the exterior magnetic field, showing further improving neurogenesis. The findings clarified the mechanism for magnetic cue-mediated neuronal differentiation, coupling protein corona and intracellular signal transduction.