Enhancement of ultrasonic thrombus imaging using novel liposomal bubbles targeting activated platelet glycoprotein IIb/IIIa complex-in vitro and in vivo study
Enhancement of ultrasonic thrombus imaging using novel liposomal bubbles targeting activated platelet glycoprotein IIb/IIIa complex-in vitro and in vivo study
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DOI：
10.1016/j.ijcard.2010.07.016
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发表时间：
2011-10-20
影响因子：
3.5
通讯作者：
Kikuchi, Makoto
中科院分区：
文献类型：
--
作者：
Hagisawa, Kohsuke;Nishioka, Toshihiko;Kikuchi, Makoto
背景：我们开发了含有Arg-Gly-Asp （RGD）序列肽的全氟碳含气气泡脂质体（BL），它与活化的血小板糖蛋白IIb/IIIa复合物结合。本研究的目的是研究这些靶向BL在体外和体内对超声血栓成像的增强作用。方法：制备由磷脂酰胆碱和胆固醇组成的脂质体，通过共价偶联反应连接RGD肽。超声将脂质体与全氟碳气体偶联，形成靶向BL。体外将靶向BL与全血混合，摇匀旋转使其凝固。在体内，我们给10只患有髂股动脉急性血栓闭塞的兔子施用靶向BL。使用7.5-9 MHz线性换能器和常规超声仪以及扫描电子显微镜对血栓进行成像。超声图像数字化，测量平均像素灰度级（黑= 0，白= 255）。结果：在体外，靶向BL组血栓的平均像素灰度水平显著高于对照组和非靶向BL组（93 +/- 26 vs. 58 +/- 16, 48 +/- 9, p = 0.002, n = 10）。扫描电镜显示大量靶向BL附着在血栓上。在体内，靶向BL血栓的平均像素灰度水平显著高于靶向BL给药前（33.2 +/- 6.4 vs. 24.8 +/- 8.5, p = 0.0051, n = 10）。结论：含全氟碳气体BL加RGD肽是一种新型超声造影剂，可显著增强体外和体内血栓超声显像，可用于急性血栓性血管闭塞的无创诊断。2010爱思唯尔爱尔兰有限公司版权所有。
Background: We developed perfluorocarbon gas-containing bubble liposomes (BL) with Arg-Gly-Asp (RGD) sequence-containing peptides, which bind to activated platelet glycoprotein IIb/IIIa complexes. The aim of this study was to examine the enhancing effects in ultrasonic thrombus imaging using these targeted BL in vitro and in vivo.Methods: Liposomes composed of phosphatidylcholine and cholesterol were manufactured, and RGD peptide was attached by a covalent coupling reaction. Sonication was used to conjugate liposomes and perfluorocarbon gas, which formed targeted BL. In vitro, targeted BL were mixed with whole blood, which was allowed to coagulate while being shaken and rotated. In vivo, we administered targeted BL to 10 rabbits with acute thrombotic occlusions in the ilio-femoral artery. Thrombi were imaged using a 7.5-9 MHz linear transducer and a conventional ultrasound machine, and by scanning electron microscopy. Ultrasound images were digitized, and mean pixel gray-scale level (black = 0, white = 255) was measured.Results: In vitro, mean pixel gray-scale level of the thrombi in targeted BL group was significantly higher than in control and non-targeted BL groups (93 +/- 26 vs. 58 +/- 16, 48 +/- 9, p = 0.002, n = 10). Scanning electron microscopy revealed large amounts of targeted BL attached to the thrombi. In vivo, mean pixel gray-scale level of the thrombi with targeted BL was significantly higher (33.2 +/- 6.4 vs. 24.8 +/- 8.5, p = 0.0051, n = 10) than that before targeted BL administration.Conclusions: Perfluorocarbon gas-containing BL with RGD peptide represent a novel echo contrast agent, which can markedly enhance ultrasonic thrombus imaging in vitro and in vivo, and may be useful for noninvasively diagnosing acute thrombotic vessel occlusion. (C) 2010 Elsevier Ireland Ltd. All rights reserved.