NDLI: Influence of Biological Media on the Structure and Behavior of Ferrocene-Containing Cationic Lipid/DNA Complexes Used for DNA Delivery

Content Provider	PubMed Central
Author	Golan, Sharon Aytar, Burcu S. Muller, John P. E. Kondo, Yukishige Lynn, David M. Abbott, Nicholas L. Talmon, Yeshayahu
Abstract	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.
Related Links	http://dx.doi.org/10.1021/la200450x
Ending Page	6621
Page Count	7
Starting Page	6615
File Format	PDF
ISSN	07437463
e-ISSN	15205827
Journal	Langmuir : the ACS journal of surfaces and colloids
Issue Number	11
Volume Number	27
Language	English
Publisher Date	2011-06-07
Access Restriction	Open
Subject Keyword	Spectroscopy Electrochemistry Materials Science(all) Surfaces and Interfaces Condensed Matter Physics Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Spectroscopy Medicine Condensed Matter Physics Materials Science Electrochemistry Surfaces and Interfaces

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