Ultrasound and magnetic resonance molecular imaging of atherosclerotic neovasculature with perfluorocarbon magnetic nanocapsules targeted against vascular endothelial growth factor receptor 2 in rats

The aim of the present study was to investigate the feasibility of using ultrasonography (US) and magnetic resonance (MR) for bimodal molecular imaging of atherosclerotic neovasculature with liquid perfluorocarbon magnetic nanocapsules (NCs) targeted to vascular endothelial growth factor receptor 2 (VEGFR-2). By incorporating perfluorooctyl bromide (PFOB) and superparamagnetic iron oxide (SPIO) into polylactic acid, a SPIO-embedded PFOB NC was constructed; subsequently, a VEGFR-2-targeted NC (VTNC) containing dual detectable probes was created by covalently linking a VEGFR-2 antibody onto the surface of the SPIO-embedded PFOB NC. Target specificity was verified in vitro by incubating VTNC with VEGFR-2(+) or VEGFR-2(−) endothelial cells. Rats with vulnerable plaques were assigned to receive either an injection of VTNC (Targeted group; n=8) or an injection of NC (Nontargeted group; n=8); control rats also received an injection of VTNC (Control group; n=8). US and MR imaging of the abdominal aorta were performed to detect VTNC by measuring of the ultrasonic grayscale intensity (GSI) and MR contrast-to-noise ratio (CNR) prior to and at successive time points following VTNC and NC injections. The percent positive area (PPA) of CD31(+) (PPA(CD31+)) or VEGFR-2(+) (PPA(VEGFR-2+)) expression was quantified by immunohistochemical staining. CD31 was used to verify the existence of endothelial cells as it is widely expressed on the surface of endothelial cells whether activated or not. The results demonstrated that VTNC was able to highly and selectively detect VEGFR-2(+) endothelial cells, and GSI, CNR, PPA(CD31+) and PPA(VEGFR-2+) were significantly increased in the targeted group compared with the nontargeted and control groups. In the control group, no atherosclerotic plaques or angiogenesis was identified, thus no expression of PPACD31+ and PPAVEGFR-2 (data not shown). There were strong correlations among GSI, CNR, PPA(CD31+) and PPA(VEGFR-2+). In conclusion, two-probe VTNC is feasible for bimodal US and MR molecular imaging of atherosclerotic neovasculature, which may offer complementary information for the more reliable prediction of plaque vulnerability.