Many studies have shown that substrate topography and mechanical strains affect various cellular activities in most cell types. However, very few studies have shown the comparative analysis of the substrate topography features with different shape and size or the combined effect of substrate topography and mechanical strain on cellular activities in neuron cells. This study investigates the effect of micro/nanopillar and pore substrate topography features and the combined effect of substrate topography and mechanical strain on neurite development in PC12 (neuron) cells. Cells were cultured on substrates with nanopillars, nanopores and micro-island features to determine the effect of substrate topography on neurite development. Similarly, cells were cultured on deformable micro-textured substrates under no strain (static) and strain levels of 4%, 8% and 16% at strain rates of 0.1Hz, 0.5Hz and 1.0Hz, to determine the effect of substrate topography and mechanical strain on neurite development. It was found that micro-islands enhanced neurite development while nanopillars enhanced proliferation compared with smooth substrates. Cells on nanopores had intermediate neurite development and proliferation. These findings suggest that the dimension of the substrate topography did affect neurite development. Under a mechanical environment, micro-textured substrates enhanced neurite development only in the static and 4% at 0.1Hz conditions. Also, on smooth and micro-textured substrates, at a lower strain level increasing the strain rate promoted enhanced neurite development while at a higher strain level high strain rate inhibited neurite development compared to their static counterparts. These findings suggest that strain level and strain rate have a combined effect on neurite development, and the effect of micro-texture on neurite development is more prominent in static and lower strain conditions than in higher strain conditions. The results of this study suggest that substrate topography and mechanical strain can be used to control neurite development in PC12 cells. These findings suggest using substrate topography and mechanical strain to control differentiation and proliferation in other neuron cell lines and neural progenitor cells. This will be therapeutically significant for treating degenerative diseases like Parkinsons disease, multiple sclerosis, spinal injury and other neuron related diseases.