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A study conducted in vivo, ex vivo, and in vitro showed that TMAO upregulated the expression of bone-associated molecules including Runx2 (Runt-related transcription factor 2) and BMP2 (bone morphogenetic protein 2), indicating that TMAO promoted osteogenic differentiation of vascular smooth muscle cells and that TMAO positively regulated vascular calcification, an effect that was mediated by TMAO through activation of NLRP3 inflammasome and NF-B signaling to promote vascular calcification