Engineered 3D Microvascular Networks for the Study of Ultrasound-Microbubble-Mediated Drug Delivery
Engineered 3D Microvascular Networks for the Study of Ultrasound-Microbubble-Mediated Drug Delivery
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DOI：
10.1021/acs.langmuir.8b03288
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发表时间：
2019-08-06
期刊：
影响因子：
3.9
通讯作者：
Averkiou, Michalakis
中科院分区：
文献类型：
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作者：
Juang, Eric K.;De Cock, Ine;Averkiou, Michalakis
局部和靶向的药物递送可以通过超声和微泡对肿瘤微环境的联合作用来实现，可能是通过声孔效应和其他尚不清楚的治疗机制。在这里，我们提出了一个灌注在体外模型与现实的三维几何形状，研究微泡和血管内皮之间的相互作用，在超声的存在。具体而言，一个三维的，内皮细胞接种在体外微血管模型灌注细胞培养基和微泡，同时由一个单一的元素1 MHz聚焦换能器进行超声处理。这种设置模拟了体内情况，其中超声在存在流动的情况下在肿瘤脉管系统中诱导治疗效果。采用荧光和明视野显微镜来评估微泡-血管相互作用以及在治疗期间和治疗后的真实的时间内药物递送和细胞死亡的程度。使用碘化丙啶作为模型药物，而使用钙黄绿素AM来评估细胞活力。有两个声学参数选择这项工作：（1）声压：1.4 MPa，脉冲长度：500个周期，占空比：5%和（2）声压：0.4 MPa，脉冲长度：1000个周期，占空比：20%。在两种情况下都观察到增强的药物递送和细胞死亡，而较高的压力设置具有更显著的效果。通过将生理流动引入体外微血管模型并检查其中内皮细胞的PECAM-1表达，我们证明了我们的模型是体内脉管系统的良好模拟物，因此是一个可行的平台，以提供对超声介导的药物递送的机制见解。
Localized and targeted drug delivery can be achieved by the combined action of ultrasound and micro-bubbles on the tumor microenvironment, likely through sonoporation and other therapeutic mechanisms that are not well understood. Here, we present a perfusable in vitro model with a realistic 3D geometry to study the interactions between microbubbles and the vascular endothelium in the presence of ultrasound. Specifically, a three-dimensional, endothelial-cell seeded in vitro microvascular model was perfused with cell culture medium and microbubbles while being sonicated by a single-element 1 MHz focused transducer. This setup mimics the in vivo scenario in which ultrasound induces a therapeutic effect in the tumor vasculature in the presence of flow. Fluorescence and bright-field microscopy were employed to assess the microbubble-vessel interactions and the extent of drug delivery and cell death both in real time during treatment as well as after treatment. Propidium iodide was used as the model drug while calcein AM was used to evaluate cell viability. There were two acoustic parameter sets chosen for this work: (1) acoustic pressure: 1.4 MPa, pulse length: 500 cycles, duty cycle: 5% and (2) acoustic pressure: 0.4 MPa, pulse length: 1000 cycles, duty cycle: 20%. Enhanced drug delivery and cell death were observed in both cases while the higher pressure setting had a more pronounced effect. By introducing physiological flow to the in vitro microvascular model and examining the PECAM-1 expression of the endothelial cells within it, we demonstrated that our model is a good mimic of the in vivo vasculature and is therefore a viable platform to provide mechanistic insights into ultrasound mediated drug delivery.