Specific Targeting of Human Inflamed Endothelium and In Situ Vascular Tissue Transfection by the Use of Ultrasound Contrast Agents
Specific Targeting of Human Inflamed Endothelium and In Situ Vascular Tissue Transfection by the Use of Ultrasound Contrast Agents
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DOI：
10.1016/j.jcmg.2009.04.012
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发表时间：
2009-08-01
影响因子：
14
通讯作者：
Sanchez-Madrid, Francisco
中科院分区：
文献类型：
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作者：
Barreiro, Olga;Aguilar, Rio J.;Sanchez-Madrid, Francisco
我们使用人脐带段作为离体模型来研究微泡（MB）的可能的临床诊断和治疗应用。一些研究已经解决了在体内应用的MBs特异性靶向血管功能障碍或声致穿孔的动物模型，但迄今为止没有人的组织models.METHODS原代微静脉内皮细胞单层靶向与MBs结合的抗体对一个高表达的内皮标志物（四跨膜蛋白CD 9），并在增加的流速（0.5至5达因/cm（2））下评估结合。此外，在离体人大血管模型（脐带静脉）中，通过对比增强超声分析在流动条件下测量CD 9偶联MB内皮靶向，并使用相同的组织模型检测具有抗细胞间粘附分子（ICAM）-1包被的MB的发炎脉管系统。最后，编码荧光蛋白的质粒被sonoporated到脐带vessels.RESULTS特定的内皮细胞靶向在体外和体外模型先前描述的是通过使用MB覆盖的抗CD 9。此外，我们设法在脐带静脉中诱导炎症，并使用抗ICAM-1偶联的MB通过实时回波成像检测炎症。此外，在该人离体模型中评估绿色荧光蛋白和绿色荧光蛋白标记的ICAM-1的表达和正确定位，而不引起血管损伤。我们已经开发了一种新的使用脐带的离体人模型，该模型对于炎症的检测和通过声致穿孔的蛋白质的外源表达是有效的。（美国科尔心脏病学杂志2009;2：997-1005）（C）2009年美国心脏病学会基金会
OBJECTIVES We used human umbilical cord segments as an ex vivo model to investigate the possible clinical diagnostic and therapeutic applications of microbubbles (MBs).BACKGROUND Microbubbles are commonly used in clinical practice as ultrasound contrast agents. Several studies have addressed the in vivo applications of MBs for specific targeting of vascular dysfunction or sonoporation in animal models, but to date no human tissue model has been established.METHODS Primary venular endothelial cell monolayers were targeted with MBs conjugated to an antibody against a highly expressed endothelial marker (tetraspanin CD9), and binding was assessed under increasing flow rates (0.5 to 5 dynes/cm(2)). Furthermore, CD9-coupled MB endothelial targeting was measured under flow conditions by contrast-enhanced ultrasound analysis in an ex vivo human macrovascular model (umbilical cord vein), and the same tissue model was used for the detection of inflamed vasculature with anti-intercellular adhesion molecule (ICAM)-1-coated MBs. Finally, plasmids encoding fluorescent proteins were sonoporated into umbilical cord vessels.RESULTS Specific endothelial targeting in the in vitro and ex vivo models described previously was achieved by the use of MBs covered with an anti-CD9. Furthermore, we managed to induce inflammation in umbilical cord veins and detect it with real-time echography imaging using anti-ICAM-1-coupled MBs. Moreover, expression and correct localization of green fluorescent protein and green fluorescent protein-tagged ICAM-1 were assessed in this human ex vivo model without causing vascular damage.CONCLUSIONS In the absence of clinical trials to test the benefits and possible applications of ultrasound contrast agents for molecular imaging and therapy, we have developed a novel ex vivo human model using umbilical cords that is valid for the detection of inflammation and for exogenous expression of proteins by sonoporation. (J Am Coll Cardiol Img 2009;2:997-1005) (C) 2009 by the American College of Cardiology Foundation