The aim of the study is to investigate the effect of mechanical stress on the enthesis. An original mechanical stress model showed the structure of the enthesis was maintained in the mild stress model. The no stress model had ill-defined borders between fibrocartilage and mineralized fibrocartilage. Calcification of mineralized fibrocartilage occurred in the strong stress model. Expression of sex-determining region Y-box 9 and scleraxis was upregulated by mild stress, whereas Runt-related transcription factor 2 and matrix metalloproteinase 13 were upregulated by strong stress. Apoptosis was inhibited by mild stress.