Flash photo stimulation of human neural stem cells on graphene/TiO2 heterojunction for differentiation into neurons

Abstract

For the application of human neural stem cells (hNSCs) in neural regeneration and brain repair, it is necessary to stimulate hNSC differentiation towards neurons rather than glia. Due to the unique properties of graphene in stem cell differentiation, here we introduce reduced graphene oxide (rGO)/TiO₂ heterojunction film as a biocompatible flash photo stimulator for effective differentiation of hNSCs into neurons. Using the stimulation, the number of cell nuclei on rGO/TiO₂ increased by a factor of ∼1.5, while on GO/TiO₂ and TiO₂ it increased only ∼48 and 24%, respectively. Moreover, under optimum conditions of flash photo stimulation (10 mW cm⁻² flash intensity and 15.0 mM ascorbic acid in cell culture medium) not only did the number of cell nuclei and neurons differentiated on rGO/TiO₂ significantly increase (by factors of ∼2.5 and 3.6), but also the number of glial cells decreased (by a factor of ∼0.28). This resulted in a ∼23-fold increase in the neural to glial cell ratio. Such highly accelerated differentiation was assigned to electron injection from the photoexcited TiO₂ into the cells on the rGO through Ti–C and Ti–O–C bonds. The role of ascorbic acid, as a scavenger of the photoexcited holes, in flash photo stimulation was studied at various concentrations and flash intensities.