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J Pharm Pharmacol. 2010 Sep;62(9):1103-11. doi: 10.1111/j.2042-7158.2010.01140.x.

Development of nanosomes using high-pressure homogenization for gene therapy.

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  • 1Center for Nanomedicine and Drug Delivery, Xavier University College of Pharmacy, New Orleans, Louisiana 70125-1098, USA.

Abstract

OBJECTIVES:

The aim of this project was to develop a novel lipid-based formulation suitable for gene therapy.

METHODS:

Novel nanosize liposome (nanosome) formulations containing pDNA (plasmid DNA) were developed using high-pressure homogenization (HPH). The effect of lipid concentration was studied at two levels: 3 mm and 20 mm. The preformed nanosomes were incubated for 18-20 h with pDNA or pDNA/protamine sulfate (PS) complex. The physical properties of the pDNA nanosomes were compared by particle size distribution and zeta-potential measurements. Their biological properties were also compared by pDNA efficiency of encapsulation/complexation, integrity, nuclease digestion, transfection efficiency and cell cytotoxicity.

KEY FINDINGS:

pDNA nanosomes prepared with 20 mM lipid (nanosomes:pDNA:PS at a ratio of 8.6:1:2) had particle sizes of 170-422 nm (90% confidence). The zeta-potential of the formulation was 49.2 +/- 1.5 mV, and the pDNA encapsulation/complexation efficiency was approximately 98%. pDNA nanosomes prepared with 3 mM lipid (nanosomes:pDNA PS at a ratio of 2.09:1:2) had particle sizes of 140-263 nm (90% confidence). The zeta-potential of this formulation was 36.4 +/- 1.2 mV, and the pDNA encapsulation/complexation efficiency was approximately 100%. However, a comparison of the efficiency of transfection indicated that pDNA nanosomes prepared with low-concentration lipids (3 mM) showed significantly higher transfection efficiency compared with the pDNA nanosomes prepared with high-concentration lipids (20 mM), as well as those prepared with Fugene-6 (a commercially available transfection reagent). This particular formulation (pDNA nanosomes, 3 mM lipids) also showed significantly less cytotoxicity compared with the other pDNA nanosome formulations.

CONCLUSIONS:

To conclude, these results indicate that condensing pDNA with PS followed by subsequent complexation with low-concentration nanosomes generated from HPH can produce a pDNA nanosome formulation that will boost transfection efficiency, while minimizing cytotoxicity. This new technology appears to be an efficient tool for future commercial or large-scale manufacture of DNA delivery systems for gene therapy.

[PubMed - indexed for MEDLINE]
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