有针对性的超声微泡销毁技术简介
Summary
超声针对性的微泡销毁（UTMD）可用于直接站点特定的生物活性分子的交付，包括基因治疗的靶器官，如心脏和肝脏超声，<sup> 1-6</sup>。
Abstract
UTMD，生物活性分子，如带负电荷的质粒DNA编码感兴趣的基因的载体，添加阳离子脂质微泡造影剂7-9炮弹。在小鼠中，这些载体携带的微泡可静脉给药，或直接向心脏的左心室。在较大的动物，他们也可以通过冠状动脉导管注入。随后的交付，从流通到靶器官，发生在微泡的共振频率的声空化。这很可能是产生机械能在短暂的或有针对性的地区的10个微血管内皮细胞之间的孔隙形成的微气泡的破坏结果。由于这种sonoporation效应的结果，和跨内皮细胞的转染效率的提高，和转基因的编码向量存放到周围组织。质粒DNA留在流通迅速降解血液中的核酸交货的可能性，从而进一步降低非超声组织和高度特异性的靶器官转。
Protocol
1。微气泡的股票准备在10毫升的PBS混合1克葡萄糖200毫克1,2 – 二棕榈酰- SN -甘油-3 – 磷脂和50毫克1,2 – 二棕榈酰- SN -甘油-3 – 磷脂。 在沸水浴20-30分钟中加热的混合物，吸管搅拌，每隔5分钟。 该解决方案可以存储在4℃至6个月。 2。微泡的制备以250μL的准备微泡造影剂原液和孵化，在40 ° 15分钟。 预热的微泡造影剂的解决方案，然后转移?…
Discussion
UTMD代表一个基因传递的新方法。作为一个平台技术，它可以与许多潜在的基因治疗策略相结合，提供无数时需要高度的组织特异性的生物活性分子。生物技术的主要限制是转染效率低。另一个重要的考虑是方便的靶器官超声，可显着减少干预骨或空气。该技术需要优化技术的基础上靶组织11以及声学的一个基本的了解，以限制组织损伤 11，提高工作效率。但是，它提供了一种廉?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
格兰特支持包括NHLBI HL080532，NHLBI HL073449，NCRR RR16453的AHA国家批准的援助奖（RVS）。特别感谢延长距离的课程设计和咨询组（DCDC），dcdcgroup.org，协助视频制作和美国能源部的教育补助金号P336C050047创办的DCDC。
Materials
| Name of the reagent/equipment | Company | Catalogue number | Comments |
|---|---|---|---|
| 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine | Sigma-Aldrich | P-5911 | component of the microbubble lipid shell |
| 1,2-dipalmitoyl-sn-glycero-3-phosphatidylethanolamine | Sigma-Aldrich | P-3275 | component of the microbubble lipid shell |
| glucose | Sigma-Aldrich | G5400 | thought to stabilize the microbubbles |
| phosphate-buffered saline | Sigma-Aldrich | P5368 | |
| glycerol | Sigma-Aldrich | G5516 | believed to prevent microbubbles from coalescing |
| Octafluoropropane gas | AirGas | N/A | inert gas used in clinical applications |
| VialMix dental amalgamator | Bristol-Myers Squibb | N/A | |
| 1 MHz, 13mm, unfocused transducer | Olympus | A303S-SU | |
| 20 MHz Function/Arbitrary Waveform Generator | Agilent | 33220A | |
| Power Amplifier | Krohn-Hite Co. | Model 7500 | |
| Hydrophone | Bruel and Kjaer | Type 1803 | |
| Charge Amplifier | Bruel and Kjaer | Type 2634 | |
| 500 MHz Oscilloscope | LeCroy | 9354L | |
| VisualSonics’ Vevo 2100 Imaging System with 34 MHz transducer | VisualSonics | 2100 | |
| 27G one inch tail vein catheters | VisualSonics | N/A | |
| Genie Plus infusion pump | Kent Scientific | GENIE |
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Cite This Article
Walton, C. B., Anderson, C. D., Boulay, R., Shohet, R. V. Introduction to the Ultrasound Targeted Microbubble Destruction Technique. J. Vis. Exp. (52), e2963, doi:10.3791/2963 (2011).