The present work describes the development of biomimetic materials for osteochondral tissue substitution and repair, which can be the start for a revolution in the classical procedures of orthopedic surgery. Performing biomineralization process, we succeeded to prove that biological system store and process information at the molecular level. The substitute consist in three layers: the lower layer, mimicking bone tissue, is formed by HA/Collagen (70/30)wt; the intermediate layer, mimicking the tade-mark, is formed by HA/Collagen (40/60)wt; and the upper layer, mimicking the cartilagineous layer, is formed by Collagen containing Hyaluronic Acid. In vivo tests performed on small and large size animals should that: the bony portion well integrate and gradually are reabsorbed in contact with femoral bone in rabbit. The osteochondral substitute showed the ability to repair defect in osteochondral lesion opened in horses knee and differently stimulate cells thanks to the different chemico-morphological-microstructural features of the scaffold layer.