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Development of a percutaneous optical imaging system for tracking vascular gene expression: an ultrasound-guided ex vivo feasibility study

机译:跟踪血管基因表达的经皮光学成像系统的开发:超声引导下的离体可行性研究

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Vascular gene therapy is an exciting approach for the cure and mitigation of atherosclerosis and related cardiovascular diseases. Monitoring transgene expression using noninvasive imaging techniques is a necessary complement for the success of clinical gene therapy. Optical imaging based on fluorescence signal detection from biomarker genes holds promise in the area. Green fluorescent protein (GFP) is one of the most commonly used biomarkers for the purpose. We have developed a novel percutaneous optical imaging system to track fluorescent marker expression from vasculatures following GFP gene transfer. To investigate the performance of the percutaneous system we performed experiments on porcine meat slabs. A polyethylene capillary tube was used to mimic the vessel geometry beneath a tissue layer. Under ultrasound-guidance, we placed the percutaneous optical probe nearby the 'vessel'. After obtaining control optical images of the 'vessel' filled with phosphate buffered saline (PBS), we imaged the same tube again, in which the PBS was replaced by human 293T cells transduced with GFP-carrying lentivirus. The results showed higher bright signal detected for the tube with GFP/sup +/ cells than that with PBS. This study demonstrates the capability of the system and provides insights for improvements of the current system.
机译:血管基因疗法是治疗和减轻动脉粥样硬化及相关心血管疾病的令人兴奋的方法。使用无创成像技术监测转基因表达是临床基因治疗成功的必要补充。基于生物标志物基因荧光信号检测的光学成像技术在该领域具有广阔的前景。为此目的,绿色荧光蛋白(GFP)是最常用的生物标记之一。我们已经开发了一种新型的经皮光学成像系统,以跟踪GFP基因转移后来自脉管系统的荧光标记表达。为了研究经皮系统的性能,我们在猪的肉板上进行了实验。使用聚乙烯毛细管模拟组织层下面的血管几何形状。在超声引导下,我们将经皮光学探头放在“血管”附近。在获得充满磷酸盐缓冲盐水(PBS)的“血管”的控制光学图像后,我们再次对同一管成像,其中PBS被载有GFP慢病毒的人293T细胞替代。结果显示,使用GFP / sup + /细胞的试管检测到的亮信号要比使用PBS的试管高。这项研究演示了系统的功能,并为改进当前系统提供了见识。

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