Contrast agent-free sonoporation: The use of an ultrasonic standing wave microfluidic system for the delivery of pharmaceutical agents
Contrast agent-free sonoporation: The use of an ultrasonic standing wave microfluidic system for the delivery of pharmaceutical agents
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DOI：
10.1063/1.3660352
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发表时间：
2011-12-01
期刊：
影响因子：
3.2
通讯作者：
Hill, Martyn
中科院分区：
文献类型：
--
作者：
Carugo, Dario;Ankrett, Dyan N.;Hill, Martyn
超声作用是促进治疗药物从细胞外环境到细胞内环境的跨膜递送的一种有用的生物物理机制。传统上，超声造影剂是在超声造影剂存在的情况下进行的，众所周知，超声造影剂可以极大地增强生物细胞膜的瞬时穿孔。然而，在体内已经观察到造影剂会由于内皮细胞的损伤而导致毛细血管破裂和出血，并极大地增加体外细胞裂解的可能性。在这里，我们使用一种低成本的超声-微流控装置展示了在没有造影剂的情况下对心肌成肌细胞进行的声旋转术(无CA声旋转术)。在这个装置中产生了超声波驻波，允许控制细胞的位置和声辐射力的强度。实时单细胞分析和超声辐照后的回顾性分析表明，无CA的超声处理可诱导荧光探针(CMFDA和FITC-葡聚糖)的跨膜转移，不同的机制可能有助于在超声波存在的情况下进行膜穿孔。此外，据我们所知，我们首次表明，由于无CA超声促进药物(阿霉素、木犀草素和芹菜素)的摄取，声波切割可导致细胞毒性增加。这里设计的美国微流控装置提供了一种体外替代昂贵和有争议的体内模型的方法，用于早期药物发现、药物输送计划和毒性测量。(C)2011年美国物理研究所。[DOI：10.1063/1.3660352]
Sonoporation is a useful biophysical mechanism for facilitating the transmembrane delivery of therapeutic agents from the extracellular to the intracellular milieu. Conventionally, sonoporation is carried out in the presence of ultrasound contrast agents, which are known to greatly enhance transient poration of biological cell membranes. However, in vivo contrast agents have been observed to induce capillary rupture and haemorrhage due to endothelial cell damage and to greatly increase the potential for cell lysis in vitro. Here, we demonstrate sonoporation of cardiac myoblasts in the absence of contrast agent (CA-free sonoporation) using a low-cost ultrasound-microfluidic device. Within this device an ultrasonic standing wave was generated, allowing control over the position of the cells and the strength of the acoustic radiation forces. Real-time single-cell analysis and retrospective post-sonication analysis of insonated cardiac myoblasts showed that CA-free sonoporation induced transmembrane transfer of fluorescent probes (CMFDA and FITC-dextran) and that different mechanisms potentially contribute to membrane poration in the presence of an ultrasonic wave. Additionally, to the best of our knowledge, we have shown for the first time that sonoporation induces increased cell cytotoxicity as a consequence of CA-free ultrasound-facilitated uptake of pharmaceutical agents (doxorubicin, luteolin, and apigenin). The US-microfluidic device designed here provides an in vitro alternative to expensive and controversial in vivo models used for early stage drug discovery, and drug delivery programs and toxicity measurements. (C) 2011 American Institute of Physics. [doi:10.1063/1.3660352]