Basic Science Investigation
In vitro imaging study of E-selectin receptor-targetting ultrasound contrast agent
Hao Wu, Yuli Zhou, Jing Zhang, Meijun Zhou, Sushu Li, Hairong Zheng, Hongmei Liu
Published 2017-02-25
Cite as Chin J Nucl Med Mol Imaging, 2017, 37(2): 86-90. DOI: 10.3760/cma.j.issn.2095-2848.2017.02.005
Abstract
ObjectiveTo develop a vulnerable plaque targeting ultrasound contrast agent (UCA) and to evaluate its affinity and imaging performance in vitro.
MethodsE-selectin receptor-targeting UCA, which conjugated with monoclonal antibody of E-selectin, was prepared with filming-rehydration method and biotin-avidin linkage. The size and distribution of UCA were measured with particle size analyzer, the connectivity condition of microbubbles with E-selectin antibody was also detected with fluorescence analysis. The cytotoxicity from microbubble and ultrasound irradiation was evaluated through cell counting kit-8 (CCK8) assay. The adhesion effect of UCA was assessed after co-incubated with activated mouse endothelial cells (bEnd.3) and compared with that of free antibody intervention group and control group. The imaging performance of UCA at different time points was observed on an ultrasound equipment with a high-frequency transducer. Two-sample t test and one-way analysis of variance were performed to analyze the data.
ResultsE-selectin receptor-targeting UCA was successfully prepared. The cytotoxicity result with CCK8 assay demonstrated the favorable biocompatibility of UCA. The connection amount of UCA on activated bEnd.3 cells ((6.23±0.45) bubbles/cell) was significantly higher than that of the free antibody intervention group ((1.57±0.34) bubbles/cell) and control group ((0.07±0.03) bubbles/cell; F=291.43, P<0.01). The performance of in vitro ultrasonography at the same time points showed no obvious difference between targeting UCA and control UCA (all t<0.51, all P>0.05).
ConclusionsThe prepared E-selectin receptor-targeting UCA has favorable targeting and imaging capabilities. It might be a potentially ultrasound molecular imaging agent for early detection and prognosis evaluation of vulnerable plaque.
Key words:
E-selectin; Contrast media; Microbubbles; Ultrasonography; Endothelial cells; Mice
Contributor Information
Hao Wu
Department of Ultrasonic Medicine, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China
Yuli Zhou
Department of Ultrasonic Medicine, Shenzhen People′s Hospital, Shenzhen 518001, China
Jing Zhang
Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
Meijun Zhou
Department of Ultrasonic Medicine, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China
Sushu Li
Department of Ultrasonic Medicine, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China
Hairong Zheng
Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
Hongmei Liu
Department of Ultrasonic Medicine, the Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China
