Naked pDNA/hyaluronic acid powder shows excellent long-term storage stability and gene expression in murine lungs

Naked pDNA/hyaluronic acid powder shows excellent long-term storage stability and gene expression in murine lungs

Takaaki Ito, Maino Fukuhara, Tomoyuki Okuda, Hirokazu Okamoto

Abstract

We evaluated the storage stability of powder containing naked plasmid DNA (pDNA) and hyaluronic acid (HA) or mannitol (Man) prepared by the spray-freeze-drying technique, through which we have reported high gene expression without any gene vectors. The powders composed of 5–10-µm porous particles and showing excellent dispersion were stored for 12 months under three storage conditions: 5 °C/Dry, 25 °C/Dry, and 25 °C/75% relative humidity. The humidified powders lost their porous shape within 1 week and were not suitable for inhalation characterization. On the other hand, the powders under dry conditions maintained high inhalation characteristics and pDNA integrity for 12 months. We administered the powders to the lungs of mice. The naked pDNA in HA powder showed significantly higher gene expression compared with that in Man powder and a pDNA-polyethylenimine complex solution. The gene expression of pDNA/HA powder was maintained for 12 months. These results suggest that powder containing naked pDNA is stable on storing under appropriate dry conditions and the naked pDNA/HA powder shows effective pulmonary gene expression.

Keywords

Plasmid DNA, Inhalation, Spray freeze-drying, Pulmonary drug delivery, Lung drug delivery, Gene delivery, Gene therapy

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