Introduction
Despite significant advances in intravascular stent technology, safe prevention of stent thrombosis over prolonged periods after initial deployment persists as a medical need to reduce device failure. The objective of this project was to assess the potential of perfluorocarbon nanoparticles conjugated with the direct thrombin inhibitor D-phenylalanyl-L-prolyl-L-arginyl chloromethylketone (PPACK-NP) to inhibit stent thrombosis.
Methods
In a static model of stent thrombosis, 3 mm × 3 mm pieces of stainless steel coronary stents were cut and adsorbed with thrombin to create a procoagulant surface that would facilitate thrombus development. Following treatment with PPACK-NP or control NP, stents were exposed to platelet poor plasma (PPP) or platelet rich plasma (PRP) for set time points up to 60 minutes. Measurements of final clot weight in grams were utilized for assessing the effect of nanoparticle treatment on limiting thrombosis. Additionally, groups of stents were exposed to flowing plasma containing various treatments (saline, free PPACK, control NP and PPACK-NP) and generated thrombi were stained and imaged to investigate the treatment effects of PPACK-NP under flow conditions.
Results
The static model of stent thrombosis utilized in this study indicated a significant reduction in thrombus deposition with PPACK-NP treatment (0.00067 ± 0.00026g, N=3) compared to control NP (0.0098 ± 0.0015g, N=3, p = 0.026) in PPP. Exposure to PRP demonstrated similar effects with PPACK-NP treatment (0.00033 ± 0.00012g, N=3) versus control NP treatment (0.0045 ± 0.00012g, N=3, p = 0.000017). In additional studies, stents were exposed to both platelet rich plasma pretreated with vorapaxar and PPACK-NP, which illustrated adjunctive benefit to oral platelet inhibitors for prevention of stent thrombosis. Additionally, an in vitro model of stent thrombosis under flow conditions established that PPACK-NP treatment significantly inhibited thrombus deposition on stents.
Conclusion
This study demonstrates that anti-thrombin perfluorocarbon nanoparticles exert marked focal anti-thrombin activity to prevent intravascular stent thrombosis and occlusion.