Optimization of Bipolar Microsecond Electric Pulses for DNA Vaccine Delivery.

Objective: Bipolar microsecond and submicrosecond pulsed electric fields have several advantages over longer duration monopolar pulses including significant reductions in muscle stimulation and perceived pain enabling their use in several novel clinical applications. In this study, treatment parameters were optimized to enhance DNA uptake in a 3D tissue model.

Methods: 3D tissue models were subjected to microsecond pulsed electric field treatments with various waveforms, doses, and delivery rates. Small molecule uptake and viability were evaluated in search of optimal outcomes. Computational models were then used to derive reversible and lethal thresholds for each treatment group. DNA transfection was then evaluated for a subset of optimal parameters and compared to traditional electroporation protocols.

Results: A 2-1-2 waveform with a 1ms dose delivered at a rate of 100μs/s resulted in the highest number of transfected cells yielding a 7730% increase over traditional monopolar pulse protocols.

Conclusion: Bipolar microsecond pulses offer substantial promise for DNA delivery via reversible electroporation.

Significance: Several gene-related therapeutics such as DNA vaccines are currently hindered by poor cellular uptake This crucial barrier to a new generation of such therapies can be overcome by improving DNA delivery as demonstrated in this work.