微流控可控分步组装制备透明质酸/聚乙烯亚胺/DNA纳米复合物

沈逸怀; 相佳佳; 申有青

doi:10.11777/j.issn1000-3304.2020.20043

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微流控可控分步组装制备透明质酸/聚乙烯亚胺/DNA纳米复合物

论文 | 更新时间：2021-01-26

- 微流控可控分步组装制备透明质酸/聚乙烯亚胺/DNA纳米复合物
- A Microfluidics-based Stepwise Assembly Method for Preparation of HA/PEI/DNA Polyplexes
- 高分子学报 2020年51卷第9期页码：1029-1038
- 作者机构：
  
  浙江大学化学工程与生物工程学院生物纳米工程中心　杭州 310027
- 作者简介：
  
  E-mail: shenyq@zju.edu.cn You-qing Shen, E-mail: shenyq@zju.edu.cn
- 基金信息：
  
  国家自然科学基金(基金号 51833008，U1501243，21704090)资助项目
- DOI：10.11777/j.issn1000-3304.2020.20043
  中图分类号：
- 纸质出版日期：2020-8-15，
  
  网络出版日期：2020-7-3，
  
  收稿日期：2020-2-24，
  
  修回日期：2020-4-20，
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沈逸怀, 相佳佳, 申有青. 微流控可控分步组装制备透明质酸/聚乙烯亚胺/DNA纳米复合物[J]. 高分子学报, 2020,51(9):1029-1038.

Yi-huai Shen, Jia-jia Xiang, You-qing Shen. A Microfluidics-based Stepwise Assembly Method for Preparation of HA/PEI/DNA Polyplexes[J]. Acta Polymerica Sinica, 2020,51(9):1029-1038.
沈逸怀, 相佳佳, 申有青. 微流控可控分步组装制备透明质酸/聚乙烯亚胺/DNA纳米复合物[J]. 高分子学报, 2020,51(9):1029-1038. DOI： 10.11777/j.issn1000-3304.2020.20043.

Yi-huai Shen, Jia-jia Xiang, You-qing Shen. A Microfluidics-based Stepwise Assembly Method for Preparation of HA/PEI/DNA Polyplexes[J]. Acta Polymerica Sinica, 2020,51(9):1029-1038. DOI： 10.11777/j.issn1000-3304.2020.20043.

摘要

阳离子聚合物/DNA形成的复合物纳米颗粒呈正电性，因此表面必须遮盖一层电中性或负电性的聚合物才能在体内应用，但如何控制遮蔽层在复合物纳米颗粒上组装，形成结构可控、尺寸均一的基因输送系统是关键. 本文以基因输送系统中常见的透明质酸(HA)/聚乙烯亚胺(PEI)/DNA系统为例，探索了利用微流控芯片进行可控分步的层层自组装，制备尺寸大小均一、表面电势为负的HA/PEI/DNA纳米复合物的方法. 将PEI与DNA通过第一个微流控芯片自组装得到PEI/DNA纳米复合物，该复合物颗粒在第二个微流控芯片内与HA再次组装得到HA/PEI/DNA纳米复合物. 考察了微流控芯片管道的尺寸、溶液流速及流速比

、PEI与DNA氮磷比(N:P)、HA与DNA质量比(HA:DNA)等参数与所形成的纳米复合物的尺寸、均一性及表面电势的关系，并与涡旋振荡法制备的复合物进行比较. 结果表明，传统的涡旋振荡法制备的HA/PEI/DNA纳米复合物尺寸偏大(340 ~ 490 nm)、均一度低(PDI

0.506 ~ 0.863)；而用微流控法制备的复合物尺寸较小(190 nm)、分布更为均一(PDI = 0.316).

Abstract

The

in vivo

applications of cationic polymer/DNA polyplexes have been limited by their positively charged surface

and thus they are generally coated with neutral or negative polymers to shield the positive charges. The key is how to control the assembly process of the shielding layer on the polyplexes surface to form a gene delivery system with controllable structure and uniform particle size. Herein

using hyaluronic acid (HA)/polyethyleneimine (PEI)/DNA gene delivery system as an example

we explored a microfluidics-based stepwise layer-by-layer assembly method for preparation of HA/PEI/DNA polyplexes with uniform particle size and negatively charged surface. PEI and DNA solutions were mixed in a microfluidic chip to form the polyplexes

which were then assembled with HA in the other microfluidic chip to obtain HA/PEI/DNA polyplexes. The parameters including microfluidic channel width

flow rates

the ratio

of the flow rates

PEI:DNA nitrogen to phosphorus ratio (N:P) and HA:DNA mass ratio (HA:DNA)

were investigated to study their relationships with the size

uniformity and surface potential of the polyplexes. Under optimum assembly conditions

HA/PEI/DNA polyplexes fabricated through microfluidic chips are much smaller (190 nm) and more uniform (PDI = 0.316) than those prepared by vortex mixing (340 − 490 nm

PDI ~ 0.506 − 0.863).

关键词

聚合物基因载体纳米复合物微流控芯片组装条件透明质酸遮蔽

Keywords

Polymeric gene carriersPolyplexesMicrofluidic chipsAssembly conditionsHA shielding layer

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