Numerical study of gene electrotransfer efficiency based on electroporation volume and electrophoretic movement of plasmid DNA.

BACKGROUND

The efficiency of gene electrotransfer, an electroporation-based method for delivery of pDNA into target tissues, depends on several processes. The method relies on application of electric pulses with appropriate amplitude and pulse duration. A careful choice of electric pulse parameters is required to obtain the appropriate electric field distribution, which not only controls the electroporated volume, but also affects the movement of pDNA. We used numerical modeling to assess the influence of different types of electrodes and pulse parameters on reversibly electroporated volume and on the extent of pDNA-membrane interaction, which is necessary for successful gene electrotransfer.

METHODS

A 3D geometry was built representing the mice skin tissue and intradermally injected plasmid volume. The geometry of three different types of electrodes (plate, finger, needle) was built according to the configuration and placement of electrodes used in previously reported in vivo experiments of gene electrotransfer. Electric field distribution, resulting from different pulse protocols was determined, which served for calculation of reversible electroporation volume and for simulation of electrophoretic movement of pDNA. The efficiency of gene electrotransfer was evaluated in terms of predicted amount of pDNA present inside the volume of reversible electroporation at the end of pulse delivery.

RESULTS

According to results of our numerical study, finger and needle electrodes provide larger amount of pDNA inside the volume of reversible electroporation than plate electrodes. However, these results are not consistent with the experiments showing that plate electrodes achieve the best transfection efficiency. Some inconsistencies were observed also by comparing the efficiencies of different high and low voltage pulse combinations, delivered by plate electrodes. The reason for inconsistencies probably lies in insufficient knowledge regarding the electroporation of stratum corneum. Namely, the size of the regions with high electrical conductivity, created by electroporation, was found to strongly affect predicted transfection efficiency.

CONCLUSIONS

The presented numerical model simulates the two most important processes involved in gene electrotransfer: electroporation of cells, and electrophoretic movement of pDNA. The inconsistencies between the model and experiments indicate incomplete knowledge of skin electroporation, or the involvement of other mechanisms, whose importance has not been yet identified.