α‐Asarone Promotes Tendon–Bone Healing Through Regulating <i>Dmp1</i> Transcription via Targeting Transcription Factor <scp>PPARG</scp> in <scp>BMSCs</scp>

The tendon-bone interface (TBI) is challenging to restore following injury, frequently resulting in unsatisfactory healing even after surgical reconstruction. α-Asarone (αASA), a bioactive ingredient derived from the Chinese medicinal plant Calamus, has shown benefits in the treatment of inflammatory conditions. However, its applications in musculoskeletal repair are rarely investigated. This was the first study to examine the therapeutic effects of αASA on TBI healing and elucidate the associated healing mechanisms. In a mouse model of TBI healing, αASA treatment significantly improved the biomechanical properties and osseointegration of tendon-bone samples over 10 weeks. The addition of αASA to in vitro cultures of bone marrow mesenchymal stem cells (BMSCs) greatly enhanced osteogenic differentiation. Using network pharmacology, 114 co-targeting genes were identified between αASA targets and TBI-related genes. RNA-seq analysis revealed that the top 20 differentially expressed genes (DEGs) were involved in tissue mineralization and ossification processes. A total of 207 transcription factors (TFs) were predicted for these DEGs, with 9 identified as core co-target genes. Surface plasmon resonance (SPR) confirmed the strong affinity of αASA for the PPARG TF, while luciferase assays demonstrated PPARG binding to the Dmp1 promoter to regulate transcription. Thus, αASA promotes osteogenic differentiation and improves TBI healing by selectively downregulating PPARG, hence reducing PPARG binding to the Dmp1 promoter. This enhances Dmp1 transcription, a critical factor in osteoblast maturation and mineralization, leading to improved tendon-bone integration. These findings provide new insights into the potential to apply αASA for enhancing TBI healing in the management of tendon-bone injuries.