Purpose Rotator cuf tendon-bone healing often leads to scarring and low biomechanical strength, resulting in a tendency to re-tear. This study examined whether combining autologous osteochondral transplantation and periosteum transplantation increases ibrocartilage transition zone regeneration and improves biomechanical ixation. Methods A total of 48 New Zealand white rabbits were divided into the periosteum, autologous osteochondral, combination of autologous osteochondral and periosteum, and control groups. The supraspinatus tendon was cut from the greater tuberosity and repaired by diferent transplants. A total of 12 rabbits were used for histological examination (haematoxylin and eosin staining, Masson's staining and Safranin-O staining) at 4, 8 and 12 weeks after the repair, and 36 rabbits were used for biomechanical tests (maximal failure load and stifness). Results At 4 weeks following the operation, each group had a large tendon-bone gap with a small number of disordered collagen ibres. At 8 weeks, the tendon-bone gap was smaller than that before the operation, and the tendon-bone gap in each experimental group was smaller with neater and denser collagen ibres and chondrocytes than in the control group, with the osteochondral combined periosteum group having the best results. At 12 weeks, the typical tendon-bone transitional structure was observed in the osteochondral combined periosteum group, and more collagen ibres and chondrocytes were generated in each group. The osteochondral combined periosteum group had the largest staining area and the largest amount of cartilage. The maximum tensile strength and stifness of each group increased over time. There was no signiicant diference in each group's maximum tensile strength and stifness at 4 weeks after the operation. However, the maximum tensile strength and stifness of the osteochondral combined periosteum group at 8 and 12 weeks after operation were signiicantly higher than those of other groups (P < 0.05). Conclusion Histological and biomechanical results show that autologous osteochondral transplantation combined with periosteum transplantation can efectively promote the regeneration of ibrous cartilage in the tendon-bone junction of the rotator cuf. It is concluded that this technique is a new treatment method to promote tendon-bone healing in the rotator cuf.