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. 2011 Nov 18;3(4):848-64.
doi: 10.3390/pharmaceutics3040848.

Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy

Behfar Moghaddam  1 , Sarah E McNeil, Qinguo Zheng, Afzal R Mohammed, Yvonne Perrie
Affiliations

Exploring the correlation between lipid packaging in lipoplexes and their transfection efficacy

Behfar Moghaddam et al. Pharmaceutics. .

Abstract

Whilst there is a large body of evidence looking at the design of cationic liposomes as transfection agents, correlates of formulation to function remain elusive. In this research, we investigate if lipid packaging can give further insights into transfection efficacy. DNA lipoplexes composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) in combination with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or 1,2-stearoyl-3-trimethylammonium-propane (DSTAP) were prepared by the lipid hydration method. Each of the formulations was prepared by hydration in dH2O or phosphate buffer saline (PBS) to investigate the effect of buffer salts on lipoplex physicochemical characteristics and in vitro transfection. In addition, Langmuir monolayer studies were performed to investigate any possible correlation between lipid packaging and liposome attributes. Using PBS, rather than dH2O, to prepare the lipoplexes increased the size of vesicles in most of formulations and resulted in variation in transfection efficacies. However, one combination of lipids (DSPE:DOTAP) could not form liposomes in PBS, whilst the DSPE:DSTAP combination could not form liposomes in either aqueous media. Monolayer studies demonstrated saturated lipid combinations offered dramatically closer molecular packing compared to the other combinations which could suggest why this lipid combination could not form vesicles. Of the lipoplexes prepared, those formulated with DSTAP showed higher transfection efficacy, however, the effect of buffer on transfection efficiency was formulation dependent.

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Figures

Figure 1.
Figure 1.
Molecular structure of DOPE and DSPE and the cationic lipids DOTAP and DSTAP used. (DOPE: 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, DSPE: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, DOTAP: 1,2-dioleoyl-3-trimethylammonium-propane, DSTAP: 1,2-stearoyl-3-trimethylammonium-propane).
Figure 2.
Figure 2.
Compression isotherm studies of the pure and mixture of lipid monolayers of DOPE:DOTAP, DSPE: DOTAP,DOPE:DSTAP and DSPE:DSTAP in deionised water or PBS at 20 °C. Results are expressed as the means of three experiments. SD has not shown for clarity.
Figure 3.
Figure 3.
DNA association within lipoplexes of DOPE:DOTAP, DOPE:DSTAP, DSPE:DOTAP in distilled water and PBS and in various concentrations of DNA. Results denote mean ± SD, n = 3.
Figure 4.
Figure 4.
Vesicle size (A) and Zeta potential (B) of DNA lipoplexes of DOPE:DOTAP, DOPE:DSTAP, DSPE:DOTAP in distilled water and PBS and in various concentrations of DNA. Results denote mean ± SD, n = 3.
Figure 5.
Figure 5.
(A) Comparison of transfection efficiency of five cationic liposomes formulated with distilled water or PBS. Results denote mean ± SD, n = 3. (B) Relative cell viability of five cationic liposomes formulated with distilled water or PBS. Results denote mean ± SD, n = 3.
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