Degradable polyelectrolyte multilayers that promote the release of siRNA.
Degradable polyelectrolyte multilayers that promote the release of siRNA.
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DOI：
10.1021/la200815t
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发表时间：
2011-06-21
期刊：
影响因子：
--
通讯作者：
Lynn DM
中科院分区：
文献类型：
--
作者：
Flessner RM;Jewell CM;Anderson DG;Lynn DM
我们报告了一种设计可降解聚电解质薄膜的方法，用于从表面控制释放siRNA。我们的方法是基于使用siRNA和可水解降解的聚（β-氨基酯）（聚合物1）逐步逐层组装多层生物膜（或“生物多层膜”，PEM）。使用用于绿色荧光蛋白（GFP）或萤火虫荧光素酶的siRNA序列制造膜导致siRNA膜（约50 nm厚）的线性生长，其在生理相关介质中孵育后促进siRNA的表面介导释放。这些含有siRNA的膜的物理化学表征显示，与使用聚合物1和较大质粒DNA构建体制造的PEM相比，膜生长曲线、物理侵蚀曲线和siRNA释放曲线存在很大差异。例如，尽管使用质粒DNA制造的膜在约48小时的时间段内逐渐侵蚀并释放DNA，但使用siRNA制造的膜在孵育的第一个小时内释放约65%的掺入的siRNA，然后发生任何观察到的膜侵蚀。在接下来的23小时内，这种初始释放爆发之后是第二个较慢的释放阶段（伴随着逐渐的膜侵蚀）。释放曲线和其他行为的这些差异可能至少部分地由siRNA和质粒DNA大小的巨大差异引起。最后，我们证明了这些膜中的siRNA以保持完整、功能性的形式释放，并且能够在体外给予哺乳动物细胞后沉默靶向蛋白质表达。本研究的结果为薄膜和涂层的设计提供了平台，所述薄膜和涂层可用于在各种基本和应用背景下（例如，用于开发新的研究工具或从薄膜涂层植入物和其他装置递送的方法）。
We report an approach to the design of degradable polyelectrolyte-based films for the controlled release of siRNA from surfaces. Our approach is based on stepwise, layer-by-layer assembly of multilayered polyelectrolyte films (or ‘polyelectrolyte multilayers’, PEMs) using siRNA and a hydrolytically degradable poly(β-amino ester) (polymer 1). Fabrication of films using siRNA sequences for green fluorescent protein (GFP) or firefly luciferase resulted in linear growth of ultrathin films (~50 nm thick) that promoted the surface-mediated release of siRNA upon incubation in physiologically relevant media. Physicochemical characterization of these siRNA-containing films revealed large differences in film growth profiles, physical erosion profiles, and siRNA release profiles as compared to PEMs fabricated using polymer 1 and larger plasmid DNA constructs. For example, whereas films fabricated using plasmid DNA erode gradually and release DNA over a period of ~48 hours, films fabricated using siRNA released ~65% of incorporated siRNA within the first hour of incubation, prior to the onset of any observed film erosion. This initial burst of release was followed by a second, slower phase of release (accompanied by gradual film erosion) over the next 23 hours. These differences in release profiles and other behaviors likely result, at least in part, from large differences in the sizes of siRNA and plasmid DNA. Finally, we demonstrate that the siRNA in these films is released in a form that remains intact, functional, and able to silence targeted protein expression upon administration to mammalian cells in vitro. The results of this investigation provide a platform for the design of thin films and coatings that could be used to localize the release of siRNA from surfaces in a variety of fundamental and applied contexts (e.g., for development of new research tools or approaches to delivery from film-coated implants and other devices).
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影响因子：
10.8
作者：
Fishbein, I;Stachelek, SJ;Levy, RJ
通讯作者：
Levy, RJ
影响因子：
--
作者：
Bechler, Shane L.;Lynn, David M.
通讯作者：
Lynn, David M.
影响因子：
29.4
作者：
DeMuth, Peter C.;Su, Xingfang;Samuel, Raymond E.;Hammond, Paula T.;Irvine, Darrell J.
通讯作者：
Irvine, Darrell J.
MAD（多药剂输送）纳米层：通过分层组装的表面涂层输送多种治疗药物。
DOI：
10.1021/la9017618
发表时间：
2009-12-15
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
--
作者：
Kim BS;Smith RC;Poon Z;Hammond PT
通讯作者：
Hammond PT
DOI：
10.1038/nrd2310
发表时间：
2007-06
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
de Fougerolles A;Vornlocher HP;Maraganore J;Lieberman J
通讯作者：
Lieberman J