Recent osteochondral scaffold strategies are required to mimic native tissue, which contains a calcified layer separated non-vascular cartilage tissue from bone tissue. It provides the optimal environment to individual tissue. One of these important roles of calcified layer is to inhibit an invasion of blood vessels from bone into cartilage. Vascular invasion could affect regenerated cartilage to be replaced with bone during repair. Therefore, it is required to prevent this process using a physical barrier in osteochondral scaffold. In this study, a triphasic scaffold with a barrier layer was developed. The barrier layer was designed to function to inhibit vascular invasion and improve cartilage repair as a calcified layer. The triphasic scaffold with a barrier layer was implanted into osteochondral defect of rabbit model. After 8 and 16 weeks, we observed that the triphasic scaffold with a barrier layer could inhibit the invasion of blood vessels from bone tissue into cartilage layer during development. However, stable hyaline cartilage failed to form. This research should be improved to investigate the implications of barrier layer after confirming to form stable cartilage first as a further work.