DNA and polylysine adsorption and multilayer construction onto cationic lipid-coated microbubbles
DNA and polylysine adsorption and multilayer construction onto cationic lipid-coated microbubbles
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DOI：
10.1021/la7009034
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发表时间：
2007-08-28
期刊：
影响因子：
3.9
通讯作者：
Ferrara, Katherine W.
中科院分区：
文献类型：
--
作者：
Borden, Mark A.;Caskey, Charles F.;Ferrara, Katherine W.
我们报告了一种新的应用程序的层层（LBL）组装技术，将多层的DNA和（PLL）到预先形成的脂质包被的微泡，以增加DNA的负载能力。我们首先测量了阳离子脂质部分和盐浓度对微泡稳定性的影响。微泡的产生和稳定性是稳健的，直到10 mM NaCl中40 mol %的阳离子脂质部分。DNA的吸附是不均匀的微泡壳，主要发生在凝聚相域。吸附的DNA的量，随后吸附的PLL，线性增加与壳中的阳离子脂质的分数。通过LbL组装方法进一步增强DNA负载，以构建DNA和PLL的多层膜（PEM）。通过zeta电位分析、荧光显微镜、紫外光谱和流式细胞术的实验结果证明PEM积聚。PEM呈现两个生长阶段，并不均匀地分布在微泡表面。通过使用五对层，微泡上的DNA负载能力增强了10倍以上。然而，PEM外壳在超声辐照期间不能防止振荡或破坏。这些结果表明，表面可以划分，使多功能，高负载超声造影剂的靶向基因治疗。
We report on a novel application of the layer-by-layer (LbL) assembly technique to attach multiple layers of DNA and (PLL) onto preformed lipid-coated microbubbles to increase the DNA loading capacity. We first measured the effects of the cationic lipid fraction and salt concentration on the microbubble stability. Microbubble production and stability were robust up to a cationic lipid fraction of 40 mol % in 10 mM NaCl. DNA adsorption was heterogeneous over the microbubble shell and occurred primarily on the condensed phase domains. The amount of adsorbed DNA, and subsequently adsorbed PLL, increased linearly with the fraction of cationic lipid in the shell. DNA loading was further enhanced by the LbL assembly method to construct polyelectrolyte multilayers (PEMs) of DNA and PLL. PEM buildup was demonstrated by experimental results from zeta potential analysis, fluorescence microscopy, UV spectroscopy, and flow cytometry. The PEMs exhibited two growth stages and were heterogeneously distributed over the microbubble surface. The DNA loading capacity onto the microbubbles was enhanced by over 10-fold by using five paired layers. However, the PEM shell did not prevent oscillation or destruction during ultrasound insonification. These results suggest that the surface can be compartmentalized to make multifunctional, high-payload ultrasound contrast agents for targeted gene therapy.