Abstract: The invention relates to a double-microenvironment three-layer bionic hydrogel scaffold and a preparation method and application thereof, which are characterized in that includes a cartilage layer GL-HPKGN, an intermediate layer GL-GMA, and a bone layer GL-HP/GMAAT; by an enzymatic crosslinking reaction based on hydroxyphenylpropionic acid (HPA), kartogenin (KGN) is grafted onto gelatin to form a cartilage-specific microenvironment in GL-HPKGN; Based on hydroxyphenylpropionic acid (HPA), a dual crosslinking network was formed by enzyme crosslinking reaction and methacryloyl-glycidyl dimethicone (GMA) photo-crosslinking reaction, which was used to graft atorvastatin (AT) onto gelatin, forming GL-HP/GMAAT with bone-specific microenvironment; the intermediate layer GL-GMA was beneficial for forming a clear and defined cartilage-bone integrated structure. The introduction of the tide-line intermediate layer GL-GMA facilitated the formation of well-defined chondro-bone integrated structures.