An ultrasound contrast agent targeted to P-selectin detects activated platelets at supra-arterial shear flow conditions.
An ultrasound contrast agent targeted to P-selectin detects activated platelets at supra-arterial shear flow conditions.
复制标题
DOI：
10.1097/rli.0b013e3181ed1b3b
复制
发表时间：
2010-10
影响因子：
6.7
通讯作者：
Klibanov AL
中科院分区：
文献类型：
--
作者：
Guenther F;von zur Muhlen C;Ferrante EA;Grundmann S;Bode C;Klibanov AL
使用天然选择素配体sialyl Lewisa评价微泡造影剂在高剪切流下对血小板的靶向作用。将生物素化的聚丙烯酰胺Sialyl Lewisa或生物素化的无碳水化合物聚合物（用作对照）通过链霉亲和素接头连接至生物素化的微泡。分离活化的人血小板并附着于纤维蛋白原包被的培养皿。不含血小板的纤维蛋白原包被的培养皿或被抗P-选择素抗体封闭的血小板培养皿用作阴性对照底物。用重组P-选择素包被的培养皿作为阳性对照底物。在倒置平行板流动室中通过显微镜评估微泡粘附，壁剪切应力值为40、30、20、10和5达因/cm 2。将结合和通过微泡的比率定义为捕获效率。在每个剪切速率下，两组之间的流体流动中的微泡数量没有显著差异。唾液酸化Lewisa靶向的微泡在包括40达因/cm 2的所有流动条件下在活化的血小板和P-选择素包被的培养皿的表面上结合并缓慢滚动。靶向微泡对活化血小板和重组P-选择素的捕获效率随着剪切流的增加而降低：在5达因/cm 2下，对活化血小板的捕获效率为16.11%，对P-选择素的捕获效率为21.83%，并且在40达因/cm 2下，两组的粘附效率仍为3.4%。唾液酸化Lewisa靶向微泡与对照基质无显著粘附，对照微泡与活化血小板或重组P-选择素也无显著粘附。使用唾液酸化Lewisa，P-选择素的快速结合配体的微泡靶向，是一种有前途的策略，用于在高剪切应力条件下结合活化血小板的超声造影剂的设计。
To evaluate targeting of a microbubble contrast agent to platelets under high shear flow using the natural selectin ligand sialyl Lewisa. Biotinylated polyacrylamide Sialyl Lewisa or biotinylated carbohydrate-free polymer (used as a control) were attached to biotinylated microbubbles via a streptavidin linker. Activated human platelets were isolated and attached to fibrinogen-coated culture dishes. Fibrinogen-coated dishes without platelets or platelet dishes blocked by an anti-P-selectin antibody served as negative control substrates. Dishes coated by recombinant P-selectin served as a positive control substrate. Microbubble adhesion was assessed by microscopy in an inverted parallel plate flow chamber, with wall shear stress values of 40, 30, 20, 10 and 5 dynes/cm2. The ratio of binding and passing microbubbles was defined as capture efficiency. There was no significant difference between the groups regarding the number of microbubbles in the fluid flow at each shear rate. Sialyl Lewisa-targeted microbubbles were binding and slowly rolling on the surface of activated platelets and P-selectin-coated dishes at all the flow conditions including 40 dynes/cm2. Capture efficiency of targeted microbubbles to activated platelets and recombinant P-selectin decreased with increasing shear flow: at 5 dynes/cm2, capture efficiency was 16.11% on activated platelets vs. 21.83 % on P-selectin, and, at 40 dynes/cm2, adhesion efficiency was still 3.4 % in both groups. There was neither significant adhesion of Sialyl Lewisa-targeted microbubbles to control substrates, nor adhesion of control microbubbles to activated platelets or to recombinant P-selectin. Microbubble targeting using sialyl Lewisa, a fast-binding ligand to P-selectin, is a promising strategy for the design of ultrasound contrast binding to activated platelets under high shear stress conditions.