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Here, an automated electrospinning method is used to make fibers with different diameters by changing the concentrations of polycaprolactone\/gelatine solutions. The impact of fiber diameter, orientation, and the inclusion of superparamagnetic iron oxide nanoparticles on neural stem cell (NSC) behavior and differentiation is investigated. Fibers with an average diameter of 1189\u2009\u00b1\u2009274\u2009nm greatly improve neuronal differentiation. The highest relative expression of the neuronal marker Tuj\u20101 and the formation of interconnected neuronal networks with neurites aligned along the fiber axis demonstrate this. Incorporating SPIONS into the fibers do not damage the cells, and aligned fibers do much better than random fibers at cell proliferation, migration, and neurite alignment. 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