Gelatine hydrogels exhibit potential as biomaterials such as wound-healing materials, artificial organs, scaffolds for cell culture and drug delivery systems because of their good biocompatibility. However, their practical applications are limited by their poor mechanical properties and high degradability. In this study, mechanically fibrillated silk fibroin (fibroin nanofibers; FNF) was used to reinforce gelatine hydrogels. The resulting gelatine hydrogels with FNF exhibited enhanced toughness compared to those reinforced with conventional aqueous regenerated fibroin (RF), which were prepared by treatment with a highly concentrated LiBr solvent or a neat gelatine hydrogel while retaining their softness. The average pore size of the gelatine hydrogel was 2.2 μm, while the gelatine hydrogel containing 25 % FNF expanded to 6.7 μm. A web-like network was formed between the pores. The addition of FNF increased the relative β-sheet contents in the hydrogels to 60.3 %, suggesting that this may have caused structural changes such as increased crystallinity for gelatine-derived proteins. Furthermore, the addition of FNF inhibited the rapid enzymatic degradation of gelatine hydrogels. FNF, which can be easily prepared in water, is a safe material for both the environment and living organisms and holds promise as a biomaterial in the future.