Neutrophil Extracellular Traps Formation Adhering to Cholesterol Crystals Modulates Inflammatory Cytokines Release and Promotes Their Disruption: Insights In Situ Touch-imprints Method From Non-obstructive General Angioscopy

Both macrophages and neutrophils are recognized as innate immune cells; the former engulf cholesterol crystals (CCs) while secreting NLRP3, which also promote inflammatory responses and drive the latter into atherosclerotic plaques. Moreover, recent studies have implied a critical role of NLRP3 in neutrophil recruitment into atherosclerotic lesions as neutrophil extracellular traps (NETs), which have not been visualized in vivo adhering on CCs so far. We have observed that neutrophils simultaneously attracted by NLRP3 cause NETs formation in situ from spontaneously ruptured atherosclerotic plaques by using touch-imprints method. This study aimed to confirm by imaging that NETs react to CCs, release NLRP3-related inflammatory factor, and degenerate on the surfaces that might result in breakages. These findings raise the possibility that NET-forming neutrophils not only respond to CCs but may also contribute to their structural modification or fragmentation. Such interactions may represent a previously underappreciated bidirectional relationship between CCs and NETs in human atherothrombosis.