NLRP3‐Dependent Crosstalk between Pyroptotic Macrophage and Senescent Cell Orchestrates Trauma‐Induced Heterotopic Ossification During Aberrant Wound Healing

Abstract Heterotopic ossification (HO) represents an unwanted ossific wound healing response to the soft tissue injury which caused catastrophic limb dysfunction. Recent studies established the involvement of inflammation and cellular senescence in the tissue healing process, though their role in HO still remained to be clarified. Here, a novel crosstalk where the pyroptotic macrophages aroused tendon‐derived stem cells (TDSCs) senescence is revealed to encourage osteogenic healing during trauma‐induced HO formation. Macrophage pyroptosis blockade reduces the senescent cell burden and HO formation in NLRP3 knockout mice. Pyroptosis‐driven IL‐1β and extracellular vesicles (EVs) secretion from macrophages are determined to motivate TDSCs senescence and resultant osteogenesis. Mechanistically, pyroptosis in macrophages enhances the exosomal release of high mobility group protein 1 (HMGB1), which directly bounds TLR9 in TDSCs to trigger morbid signaling. NF‐κB signaling is confirmed to be the converging downstream pathway of TDSCs in response to HMGB1‐containing EVs and IL‐1β. This study adds insights into aberrant regeneration‐based theory for HO formation and boosts therapeutic strategy development.