A proposed source of stem cells for nerve regeneration are dental pulp stem cells (DPSCs), based on their close embryonic origin to neurons and the ease with which DPSCs can be obtained from a donor. This study evaluated the response of human DPSCs to spider dragline silk fibers, a potential substrate material for tissue regeneration. The DPSCs' morphology and spread pattern were characterized after these cells were plated onto Nephila clavipes dragline fibers in media. In addition, the responses of two other well established cell lines, osteoblasts (7F2s), and fibroblasts (3T3s), were also studied under identical conditions. The inclusion of 3T3s and 7F2s in this study allowed for both direct comparisons to prior published work and a qualitative comparison to the morphology of the DPSCs. After twelve days, the DPSCs exhibited greater relative alignment and adherence to the spider dragline fibers than the 3T3s and 7F2s. The impact of a common sterilization method (ultraviolet light) on the spider dragline fiber surface and subsequent cell response to this modified surface was also characterized. Exposure of the silk to ultraviolet light did not have a measureable effect on cell alignment, but it did eliminate bacterial growth and changed fiber surface roughness. Spiders' exposure to stressful environments did not have an effect on silk to impair cell alignment or adhesion. Synthetic recombinant protein silk did not act as a substrate for cell adhesion or alignment but hydrogels with similar composition supported cell attachment, growth and proliferation. In all cases, natural drawn spider silk acted as an effective substrate for cellular adhesion and alignment of DPSCs and could be used in neural differentiation applications.