The role of lipid charge density in the serum stability of cationic lipid/DNA complexes.
To evaluate the role of lipid charge density in the serum stability of DOTAP-Chol/DNA complexes (lipoplexes), lipid-DNA interactions, extent of aggregation, supercoil content, and in vitro transfection efficiency of lipoplexes were investigated. In general, higher serum concentration destabilized, and increasing molar charge ratio of DOTAP to negatively charged phosphates in the DNA (DOTAP(+)/DNA(-)) stabilized lipoplexes in serum as assessed by the criteria used in this study. The increase of cholesterol content led to increased serum stability, and DOTAP:Chol (mol/mol 1:4)/DNA lipoplex with DOTAP(+)/DNA(-) ratio 4 was the most serum stable formulation of all the formulations examined, and maintained lipid-DNA interactions, did not aggregate and exhibited high in vitro transfection efficiency in 50% (v/v) serum. The increased stability of this formulation could not be explained by the decreased charge density of the lipid component. Furthermore, no single parameter examined in the study could be used to consistently predict the in vitro transfection efficiency of lipoplexes in serum. Surprisingly, no correlation between the maintenance of supercoiled DNA content and in vitro transfection efficiency was found in the study.
Zhang
Ye
Y
Department of Pharmaceutical Sciences, School of Pharmacy, C238, University of Colorado, 4200 E Ninth Ave., Denver, CO 80262, USA. ye.zhang@uchsc.edu
Anchordoquy
Thomas J
TJ
D000818:Animals; D002412:Cations / Q000097:blood / Q000737:chemistry; D002784:Cholesterol / Q000097:blood / Q000737:chemistry; D004247:DNA / Q000097:blood* / Q000737:chemistry* / Q000235:genetics; D004355:Drug Stability; D005229:Fatty Acids, Monounsaturated / Q000097:blood / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D008055:Lipids / Q000097:blood* / Q000737:chemistry*; D008081:Liposomes / Q000097:blood / Q000737:chemistry; D008156:Luciferases / Q000235:genetics; D051379:Mice; D009391:Nephelometry and Turbidimetry; D009690:Nucleic Acid Conformation; D010316:Particle Size; D000644:Quaternary Ammonium Compounds / Q000097:blood / Q000737:chemistry; D055672:Static Electricity; D013997:Time Factors; D014162:Transfection
D002412:Cations; D005229:Fatty Acids, Monounsaturated; D008055:Lipids; D008081:Liposomes; D000644:Quaternary Ammonium Compounds; D004247:DNA; D002784:Cholesterol; D008156:Luciferases; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane