声打印技术及微泡载药在超声介导的纳米颗粒细胞转染中的重要性。

Sonoprinting and the importance of microbubble loading for the ultrasound mediated cellular delivery of nanoparticles.

机构信息

Laboratory of General Biochemistry and Physical Pharmacy, Ghent Research Group on Nanomedicine, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, Ghent, Belgium.

Physics of Fluids Group, MESA+ Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, P.O. Box 217, 7500 AE, Enschede, The Netherlands.

出版信息

Biomaterials. 2016 Mar;83:294-307. doi: 10.1016/j.biomaterials.2016.01.022. Epub 2016 Jan 6.

Abstract

In the last years, research on ultrasound mediated drug delivery using microbubbles is vastly expanding. While some groups simply mix drugs and microbubbles (co-administration), other researchers have a major interest in the potential of drug-loaded microbubbles. However, today, little is known on the pros and cons of these two strategies. In this study we evaluated the delivery of nanoparticles (polystyrene nanospheres and mRNA-lipoplexes) to cells in vitro, in case the nanoparticles were mixed with unloaded microbubbles versus loaded onto the microbubbles. Flow cytometry experiments demonstrated that unloaded microbubbles did not enhance the cellular delivery of the nanospheres and mRNA-lipoplexes. However, upon loading the nanoparticles onto the microbubbles, their delivery to cells substantially improved. Real-time swept field confocal microscopy imaging of the microbubbles and cells during ultrasound radiation revealed that nanoparticle-loaded microbubbles directly deposited the nanoparticles in patches onto the cell membrane, a process that we termed 'sonoprinting'. This phenomenon resulted in the delivery of large amounts of nanoparticles to the cells and is suggested to be different from the creation of cell membrane pores and enhanced endocytosis, which have been reported before as mechanisms behind the improved delivery of drugs to cells by ultrasound.

摘要

在过去的几年中,利用微泡的超声介导药物输送的研究正在迅速扩展。虽然有些小组只是将药物和微泡简单地混合(联合给药),但其他研究人员对载药微泡的潜力非常感兴趣。然而,今天,对于这两种策略的优缺点知之甚少。在这项研究中,我们评估了纳米颗粒(聚苯乙烯纳米球和 mRNA 脂质体)向体外细胞的输送情况,纳米颗粒是与未负载的微泡混合还是负载在微泡上。流式细胞术实验表明,未负载的微泡不会增强纳米球和 mRNA 脂质体的细胞内输送。然而,当将纳米颗粒加载到微泡上时,它们向细胞的输送大大改善。在超声辐射期间对微泡和细胞进行实时扫场共聚焦显微镜成像显示,载有纳米颗粒的微泡直接将纳米颗粒以斑块的形式沉积在细胞膜上,我们将这一过程称为“声打印”。这种现象导致大量纳米颗粒被输送到细胞中,并且与以前报道的超声增强药物向细胞输送的机制不同,该机制是通过创建细胞膜孔和增强内吞作用来实现的。

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