Creating substrates mimicking the extracellular matrix (ECM) is a promising approach to better control cell-material interactions. The aim of this research is to use collagen, a major component of the ECM, to elaborate nanostructured biointerfaces. Different strategies were implemented and are reviewed here. First, chemically patterned substrates were used to confine collagen assembly in given domains. Then, layer-by-layer (LbL) assembly was used to incorporate collagen in thin multilayered films, using assembly with either poly(styrene sulfonate) or fibronectin, another ECM component involved in cell adhesion. Finally, collagen-based nanotubes were synthesized using LbL assembly within the pores of a membrane. Electrophoretic deposition was used to immobilize these nanotubes at the surface of indium-tin oxide-coated glass, and preosteoblast cells were shown to interact with the nanotubes. Taken together, these methods improve our ability to direct collagen adsorption, thereby producing tailored biointerfaces for applications in biomaterials science and tissue engineering.