Abstract
We aimed to investigate the mechanism of paclitaxel-eluting stent (PES) induced thrombosis by real-time dynamic monitoring of live endothelial cells. PES is widely used in clinics to inhibit restenosis effectively. However, late stent thrombosis is a major concern with the use of PES. We established an in vitro cell culture system to mimic the close contact of endothelial cells with PES struts. The dynamic response of porcine innominate artery endothelial cells (PIECs) to stents was observed using a bright field microscope and a high-resolution charge-coupled device PES changed elongated PIECs to round PIECs within 24 hours. Paracellular gaps were readily observed in a PIEC monolayer exposed to PES. By contrast, paracellular gaps were almost undetectable in an endothelial monolayer incubated with bare metal stent (BMS). As incubation time was prolonged (days 5-9), round PIECs returned to their elongated shape, but paracellular gaps were retained at lower frequencies and smaller sizes than those on days 1 and 2. In addition, the PIEC monolayer in the PES group retained an uneven surface topology during incubation, whereas PIECs in the BMS group developed a smooth surface of epithelial cell sheets on days 5-7. Our findings showed that the shift of cell shape causes impaired integrity of the monolayer characteristic with enlarged paracellular gaps and uneven surface topology in exposure to PES. The results might serve as structural information for understanding the mechanism of PES-induced early and late thrombosis.
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