Due to excellent mechanical properties and biocompatibility, materials based on silk fibroin are increasingly included in advanced biomedical research and applications. However, their poor supporting properties for cell adhesion and proliferation represent limiting factors of the utilization. To eliminate this deficiency, we developed a series of phase-separation approaches allowing for tunable texturing of planar and 3D printed fibroin surfaces from nano to macro levels. The formation of surface structures presented is based on a combination of good and poor solvents, whereas no potentially problematic templates or additives, diminishing biocompatibility of the resulting material, are required. A critical factor in obtaining and scaling of the textures is control over the degree of transformation of fibroin secondary structures between prevalently amorphous Silk I and semicrystalline Silk II forms before and during surface treatment. Employing a set of optimized procedures, selectively or hierarchically structured fibroin surfaces can be prepared at the nano, micro, and macro level, which are characterized by long-term stability in physiological environments, allowing enhanced adhesion and proliferation of human keratinocytes as well as skin fibroblast cultivations.