A touch-and-go lipid wrapping technique in microfluidic channels for rapid fabrication of multifunctional envelope-type gene delivery nanodevices
A touch-and-go lipid wrapping technique in microfluidic channels for rapid fabrication of multifunctional envelope-type gene delivery nanodevices
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DOI：
10.1039/c1lc20392d
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发表时间：
2011-01-01
期刊：
影响因子：
6.1
通讯作者：
Baba, Yoshinobu
中科院分区：
文献类型：
--
作者：
Kitazoe, Katsuma;Wang, Jun;Baba, Yoshinobu
多功能包膜型基因传递纳米器件是一种很有前途的非病毒基因治疗载体。尽管补丁在长时间暴露于血液循环中仍然很强，免疫原性低，并且适合于基因靶向，但它们的制造需要劳动密集型过程。在这项工作中，开发了一种新的方法，通过在聚二甲基硅氧烷(PDMS)/玻璃微流控装置中使用Touch-and-Go脂质包裹技术来快速制造补片。在玻璃衬底上，通过将浓缩的质粒DNA核心引入具有多个脂质双层膜的微流控通道中，制备了该修补剂。微流控通道的修饰原理是基于凝聚的DNA核心与包覆的脂类双层膜之间的静电相互作用。构建的补充剂被收集到芯片外，并进行了动态光散射表征。修补剂在5min内形成，粒径分布集中在200 nm左右。Mend的大小与凝聚的DNA核心在微流控通道中的流速有很强的相关性，因此可以控制，从而为医学应用提供最佳的大小。这种方法也被证明可以同时在多个通道中制造修补件。与传统方法相比，这种芯片上制作的补丁非常简单、快速、方便和成本效益高。我们的结果有力地表明，用我们的微流控装置制备的补片具有良好的医疗应用潜力。此外，这种微流控装置制作的补片具有很大的临床应用潜力，因为该装置是可高压灭菌的，所有的制作步骤都可以在封闭的微流控通道中完成，而不会产生任何外部污染。
Multifunctional envelope-type gene delivery nanodevices (MENDs) are promising non-viral vectors for gene therapy. Though MENDs remain strong in prolonged exposure to blood circulation, have low immunogenic response, and are suitable for gene targeting, their fabrication requires labor-intensive processes. In this work, a novel approach has been developed for rapid fabrication of MENDs by a touch-and-go lipid wrapping technique in a polydimethylsiloxane (PDMS)/glass microfluidic device. The MEND was fabricated on a glass substrate by introduction of a condensed plasmid DNA core into microfluidic channels that have multiple lipid bilayer films. The principle of the MEND fabrication in the microfluidic channels is based on electrostatic interaction between the condensed plasmid DNA cores and the coated lipid bilayer films. The constructed MEND was collected off-chip and characterized by dynamic light scattering. The MEND was constructed within 5 min with a narrow size distribution centered around 200 nm diameter particles. The size of the MEND showed strong dependence on flow velocity of the condensed plasmid DNA core in the microfluidic channels, and thus, could be controlled to provide the optimal size for medical applications. This approach was also proved possible for fabrication of a MEND in multiple channels at the same time. This on-chip fabrication of the MEND was very simple, rapid, convenient, and cost-effective compared with conventional methods. Our results strongly indicated that MENDs fabricated with our microfluidic device have a good potential for medical use. Moreover, MENDs fabricated by this microfluidic device have a great potential for clinical use because the devices are autoclavable and all the fabrication steps can be completed inside closed microfluidic channels without any external contamination.