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  1. Abstract

    Following anterior cruciate ligament (ACL) reconstruction surgery, a staged repair response occurs where cells from outside the tendon graft participate in tunnel integration. The mechanisms that regulate this process, including the specific cellular origin, are poorly understood. Embryonic cells expressing growth and differentiation factor 5 (GDF5) give rise to several mesenchymal tissues in the joint and epiphyses. We hypothesized that cells from a GDF5 origin, even in the adult tissue, would give rise to cells that contribute to the stages of repair. ACLs were reconstructed inGdf5‐Cre;R26R‐tdTomato lineage tracing mice to monitor the contribution ofGdf5‐Cre;tdTom+cells to the tunnel integration process. Anterior−posterior drawer tests demonstrated 58% restoration in anterior−posterior stability.Gdf5‐Cre;tdTom+cells within the epiphyseal bone marrow adjacent to tunnels expanded in response to the injury by 135‐fold compared with intact controls to initiate tendon‐to‐bone attachments. They continued to mature the attachments yielding zonal insertion sites at 4 weeks with collagen fibers spanning across unmineralized and mineralized fibrocartilage and anchored to the adjacent bone. The zonal attachments possessed tidemarks with concentrated alkaline phosphatase activity similar to native entheses. This study established that mesenchymal cells from a GDF5 origin can contribute to zonal tendon‐to‐bone attachments within bone tunnels following ACL reconstruction.

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