In order to investigate the neural behaviors on nanostructure, we proposed the nanostructured substrates coated with different extracellular matrix proteins for culturing neuron cells and induced the neural differentiation by nerve growth factor (NGF). The artificial nanostructured surface is the nanorods fabricated on the silicon surface by the chemical etching method. Three kinds of substrates: cover glass slide, flat silicon wafer, and nanostructured wafer were treated with oxygen plasma to increase hydrophilic property, and coated with the 1% ECM gel, laminin-1 peptides, and IKVAV peptide respectively. Neuron PC12 cells, were cultured on the processed substrates and were induced to differentiate neurites with NGF. Distribution of cytoskeleton, cellular attachment, body morphology, and neuritogenesis were studied by scanning electron microscopy (SEM) and immunofluorescence assay (IFA). The results reveal that different substrates will influence the behavior of neuron differentiation.