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Abstract
Background: Plasmid DNA (pDNA) is attractive molecule for gene therapy. pDNA-targeted delivery by efficient and safe methods is required to enhance its intra-tissue bioavailability. Among non-viral methods, sonoporation has become a promising method for in-vitro and in-vivo pDNA delivery. The efficiency of non-viral delivery methods of pDNA is generally limited by the presence of serum.
Purpose: The aim of this study was to evaluate the influence of serum on in-vitro pDNA delivery using microbubble-assisted ultrasound.
Methods: The effects of a range of serum concentrations (0–50%) on efficiency of in-vitro pDNA delivery by sonoporation were determined on human glioblastoma cells. Furthermore, the influence of the serum on cell viability, membrane permeabilization, microbubble destruction, and pDNA topology were also assessed.
Results: In-vitro results showed that a low serum concentration (i.e. ≤1%) induced a significant increase in transfection level through an increase in cell viability. However, a high serum concentration (i.e. ≥5%) resulted in a significant decrease in cell transfection, which was not associated with a decrease in membrane permeabilization or loss in cell viability. This decrease in transfection level was in fact positively correlated to changes in pDNA topology.
Conclusion: Serum influences the efficiency of in-vitro pDNA delivery by sonoporation through change in pDNA topology.
Acknowledgements
The authors acknowledge C. Arnoult (GICC, UMR CNRS 6239, Tours, France) for his technical assistance in flow cytometry and Bracco Research Geneva for supplying the microbubbles. Dr. J. Butterworth professionally proofread the manuscript (Word Perfect Science, http://www.wordperfectscience.com).