Influence of biological media on the structure and behavior of ferrocene-containing cationic lipid/DNA complexes used for DNA delivery.
Influence of biological media on the structure and behavior of ferrocene-containing cationic lipid/DNA complexes used for DNA delivery.
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DOI：
10.1021/la200450x
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发表时间：
2011-06-07
期刊：
影响因子：
--
通讯作者：
Talmon Y
中科院分区：
文献类型：
--
作者：
Golan S;Aytar BS;Muller JP;Kondo Y;Lynn DM;Abbott NL;Talmon Y
生物介质影响阳离子脂质-DNA复合物（脂质复合物）的物理化学性质，并可影响其转染细胞的能力。为了开发新的脂质体用于有效的DNA递送，必须了解含血清介质对所得脂质复合物的结构和性质的影响。然而，迄今为止，一个清晰的和一般的图片如何含血清的媒体影响的结构的lipoplexes尚未建立。一些研究表明，血清可以分解使用某些类型的阳离子脂质形成的脂质复合物，导致转染的抑制。其他研究已经证明，由其他脂质配制的脂质复合物在血清存在下是稳定的，并且能够有效地抑制细胞。在本文中，我们描述了含血清的媒体上形成的脂质复合物的氧化还原活性阳离子脂质双（n-二茂铁十一烷基）二甲基溴化铵（BFDMA）的影响。当血清浓度从0%增加到2%、5%、10%和50%（v/v）时，该lipoplex系统促进COS-7细胞中转基因表达水平的显著降低。为了了解这种转染效率降低的原因，我们使用低温透射电子显微镜（cryo-TEM）和zeta电位的测量来表征补充有0%、2%、5%、10%和50%血清的细胞培养基中的lipoplexes。低温透射电镜显示，在无血清介质中，BFDMA脂质复合物形成洋葱状，多层纳米结构。然而，培养基中血清的存在导致完整的多层脂质复合物解离。在低血清浓度（2%和5%）下，DNA线似乎与复合物分离，使脂质复合物的纳米结构被破坏。在较高的血清浓度（10%）的解离增加，和束的多层放电的主要多层复合物。相比之下，在无血清盐水（Li 2SO 4）培养基和OptiMEM细胞培养基（无血清）中表征的脂质复合物没有表现出显著的结构变化。无血清培养基（盐培养基和细胞培养基）中脂质复合物的ζ电位相似（例如，约-35mV）。有趣的是，血清的存在导致zeta电位变得不那么负（OptiMEM中约为− 20 mV，Li 2SO 4中约为− 10 mV），即使血清含有带负电荷的实体，这些实体已被证明在其他lipoplex系统中导致更负的zeta电位。zeta电位和cryo-TEM的组合测量与DNA线在血清存在下与lipoplex分离的命题一致，导致lipoplex表面的净负电荷减少。
Biological media affect the physicochemical properties of cationic lipid-DNA complexes (lipoplexes) and can influence their ability to transfect cells. To develop new lipids for efficient DNA delivery, the influence of serum-containing media on the structures and properties of the resulting lipoplexes must be understood. To date, however, a clear and general picture of how serum-containing media influences the structures of lipoplexes has not been established. Some studies suggest that serum can disintegrate lipoplexes formed using certain types of cationic lipids, resulting in inhibition of transfection. Other studies have demonstrated that lipoplexes formulated from other lipids are stable in the presence of serum and able to transfect cells efficiently. In this paper, we describe the influence of serum-containing media on lipoplexes formed using the redox-active cationic lipid bis(n-ferrocenylundecyl)dimethylammonium bromide (BFDMA). This lipoplex system promotes markedly decreased levels of transgene expression in COS-7 cells as serum concentrations are increased from 0% to 2%, 5%, 10%, and 50% (v/v). To understand the cause of this decrease in transfection efficiency, we used cryogenic transmission electron microscopy (cryo-TEM) and measurements of zeta potential to characterize lipoplexes in cell culture media supplemented with 0%, 2%, 5%, 10%, and 50% serum. Cryo-TEM revealed that in serum-free media BFDMA lipoplexes form onion-like, multilamellar nanostructures. However, the presence of serum in the media caused disassociation of the intact multilamellar lipoplexes. At low serum concentrations (2% and 5%), DNA threads appeared to separate from the complex, leaving the nanostructure of the lipoplexes disrupted. At higher serum concentration (10%) disassociation increased, and bundles of multilamellae were discharged from the main multilamellar complex. In contrast, lipoplexes characterized in serum-free aqueous salt (Li2SO4) medium and in OptiMEM cell culture medium (no serum) did not exhibit significant structural changes. The zeta potentials of lipoplexes in serum-free media (salt medium and cell culture medium) were similar (e.g., approximately −35mV). Interestingly, the presence of serum caused zeta potentials to become less negative (about −20mV in OptiMEM and −10mV in Li2SO4), even though serum contains negatively charged entities that have been demonstrated to lead to more negative zeta potentials in other lipoplex systems. The combined measurements of zeta potential and cryo-TEM are consistent with the proposition that DNA threads separate from the lipoplex in the presence of serum, resulting in a decrease in the net negative charge of the surface of the lipoplex.
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影响因子：
5.1
作者：
Li, S;Tseng, WC;Huang, L
通讯作者：
Huang, L
影响因子：
5.1
作者：
Yang, JP;Huang, L
通讯作者：
Huang, L
DOI：
10.1098/rsta.2006.1841
发表时间：
2006-10-15
影响因子：
5
作者：
Safinya, Cyrus R.;Ewert, Kai;Samuel, Charles E.
通讯作者：
Samuel, Charles E.
影响因子：
3.9
作者：
Marchini, Cristina;Montani, Maura;Caracciolo, Giulio
通讯作者：
Caracciolo, Giulio
影响因子：
10.8
作者：
Jewell, CM;Hays, ME;Lynn, DM
通讯作者：
Lynn, DM