Barreiro Olga, Aguilar Río J, Tejera Emilio, Megías Diego, de Torres-Alba Fernando, Evangelista Arturo, Sánchez-Madrid Francisco
Servicio de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain.
JACC Cardiovasc Imaging. 2009 Aug;2(8):997-1005. doi: 10.1016/j.jcmg.2009.04.012.
We used human umbilical cord segments as an ex vivo model to investigate the possible clinical diagnostic and therapeutic applications of microbubbles (MBs).
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.
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.
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.
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.
我们使用人脐带段作为体外模型，研究微泡（MBs）可能的临床诊断和治疗应用。
微泡在临床实践中通常用作超声造影剂。多项研究探讨了微泡在动物模型中针对血管功能障碍或声穿孔的体内应用，但迄今为止尚未建立人体组织模型。
用与针对高表达内皮标志物（四跨膜蛋白CD9）的抗体偶联的微泡靶向原发性小静脉内皮细胞单层，并在增加的流速（0.5至5达因/平方厘米）下评估结合情况。此外，在体外人宏观血管模型（脐带静脉）中通过超声造影分析在流动条件下测量CD9偶联的微泡对内皮的靶向作用，并使用相同的组织模型用抗细胞间粘附分子（ICAM）-1包被的微泡检测炎症血管。最后，将编码荧光蛋白的质粒声穿孔导入脐带血管。
通过使用覆盖有抗CD9的微泡，在上述体外和体内模型中实现了特异性内皮靶向。此外，我们成功地在脐带静脉中诱导炎症，并使用抗ICAM-1偶联的微泡通过实时超声成像进行检测。此外，在该人体体外模型中评估了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，且未造成血管损伤。
在缺乏临床试验来测试超声造影剂用于分子成像和治疗的益处及可能应用的情况下，我们开发了一种使用脐带的新型人体体外模型，该模型对于检测炎症和声穿孔介导的蛋白质外源表达是有效的。
