Objective      To reconstruct anterior cruciate ligament by a composite cell-tendon scaffold with a continuous and heterogeneous transitional region to mimic native ligament insertion site in rabbits and observe the outcome.        Methods      Heterogeneous cell populations with collagen gel were seeded within specific regions of decellularized rabbit Achilles tendons to fabricate a stratified scaffold containing 3 biofunctional regions to support fibrogenesis

chondrogenesis

and osteogenesis. A total of 24 male rabbits were inflicted with ACL resection in one hind limb and then followed grafted with our obtained ACL

and the other hind limb receiving sham operation served as control. In 8 weeks later

histological staining

micro-CT scanning and immunofluorescence assay were carried out to observe the growth of the graft with bone tunnel.        Results      Histological staining and immunofluorescence assay demonstrated that the cartilage-related markers

glucosamine glycolic acid and collagen 2A1

were found only in the chondrogenesis region

but bone-related protein

osteocalcin

was only in the osteogenesis region of bone tunnel

and fibrosis was remarkable for the fibrogenesis region in the joint cavity. Micro-CT scanning showed that the newly formed mineralized tissue was detected in the bone tunnels in the experimental group compared with controls. These results suggest that the ACL was reconstructed with the cell-tendon complex in 8 weeks after transportation

and the transitional architecture with ligament-fibrocartilage-bone was constructed in the ligament-bone tunnel interface.        Conclusion        Our ACL is reconstructed successfully with the cell-tendon complex after 8 weeks. The transitional architecture with ligament-fibrocartilage-bone is constructed in the ligament-bone tunnel interface

which can be applied for the regeneration of the ligament-bone interface.