Nanomaterial-Mediated Clearance of Circulating Cell-Free DNA Reduces Periodontitis-Associated Arterial Thrombosis

Abstract

Patients with periodontitis exhibit heightened susceptibility to arterial thrombosis, predisposing them to severe cardiovascular events. Gaining insights into the immune mediators linking periodontitis to arterial thrombosis is critical for developing targeted therapeutic strategies. In this study, we demonstrated that elevated plasma levels of cell-free DNA (cfDNA), a key immune mediator, were significantly associated with periodontitis severity, as indicated by increased clinical attachment loss and elevated gingival index scores. Using a combination of human clinical samples and animal models, our research identified a key mechanistic pathway: Periodontitis-induced circulating cfDNA activates Toll-like receptor 9 (TLR9) signaling, which subsequently triggers platelet activation and neutrophil extracellular trap (NET) formation to synergistically drive procoagulant responses. In vivo, periodontitis exacerbated arterial thrombus formation and compromised vascular perfusion. Hypothesizing that targeting the cfDNA–TLR9–NET axis would help mitigate periodontitis-promoted thrombosis, biodegradable nanomaterials with a cationic surface were engineered to efficiently clear circulating cfDNA. Administration of these cfDNA scavengers suppressed platelet activation and NETosis both in vitro and in vivo, reduced arterial thrombus burden, and restored vascular perfusion. These functional improvements were accompanied by attenuation of the cfDNA–TLR9–NET cascade and upregulation of vascular contractile gene signatures. These insights highlight the cfDNA–TLR9–NET axis as a potent link between oral and vascular inflammation, offering a promising therapeutic framework to mitigate arterial thrombotic risk in patients with periodontitis.