- 作者：Olga Barreiro ; PhD?/sup> ; &dagger ; ; Rí ; o J. Aguilar ; MD ; PhD&Dagger ; ; Emilio Tejera ; PhD?/sup> ; &dagger ; ; Diego Megí ; as ; PhD§ ; ; Fernando de Torres-Alba ; MD&Dagger ; ; Arturo Evangelista ; MD ; PhD&Dagger ; ; Francisco Sá ; nchez-Madrid ; PhD?/sup> ; &dagger ; ; fsanchez.hlpr@salud.madrid.org
- 关键词：ultrasound ; microbubbles ; human umbilical cord ; targeting ; inflammation ; sonoporation
- 刊名：JACC: Cardiovascular Imaging
- 出版年：2009
- 出版时间：August 2009
- 年：2009
- 卷：2
- 期：8
- 页码：997-1005
- 全文大小：1413 K
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/cm2). 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¨Ccoated 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¨CICAM-1¨Ccoupled 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.