Date of Award

Summer 8-2020

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Program/Concentration

Biomedical Engineering

Committee Director

Shu Xiao

Committee Member

Christian Zemlin

Committee Member

Chunqi Jiang

Committee Member

Patrick Sachs

Abstract

Nanosecond pulsed electric fields (nsPEFs) offer a plethora of opportunities for developing integrative technologies as complements or alternatives to traditional medicine. Studies on the biological effects of nsPEFs in vitro and in vivo have revealed unique characteristics that suggest the potential for minimized risk of complications in patients, such as the ability of unipolar nsEPs to create permanent or transient pores in cell membranes that trigger localized lethal or non-lethal outcomes without consequential heating. A more recent finding was that such responses could be diminished by applying a bipolar pulse instead, a phenomenon dubbed bipolar cancellation, paving the way for greater flexibility in nsPEF application design. Transitioning nsPEFs into practical use, however, has been hampered by both device design optimization and the intricacies of mammalian biology. Generating electric fields capable of beneficially manipulating human physiology requires high-voltage electrical pulses of nanosecond duration (nsEPs) with high repetition rates, but pulse generator and electrode design in addition to the complex electrical properties of biological fluids and tissues dictate the strength range and distribution of the resulting electric field. Faced with both promising and challenging aspects to producing a biomedically viable option for inducing a desired nsPEF response that is both focused and minimally invasive, the question becomes: how can the distinct features of unipolar and bipolar nsPEF bioeffects be exploited in a complex electrode exposure system to spatially modulate cell permeabilization?

This dissertation presents a systematic study of an efficient coplanar quadrupole electrode nsPEF delivery system that exploits unique differences between unipolar and bipolar nsPEF effects to validate its ability to control cell responses to nsPEFs in space. Four specific aims were established to answer the research question, with specific attention to the roles played by pulse polarity, grounding configuration and electric field magnitude in influencing nsPEF stimulation of electropermeabilization in space. Using a prototype wire electrode applicator charged by a custom-built multimodal pulse generator, the aims were to spatially quantifyelectropermeabilization due (1) unipolar and (2) bipolar nsPEF exposure, to (3) apply synchronized pulses with a view to canceling bipolar cancellation (CANCAN) through superposition that could shift the effective nsPEF response, and to (4) evaluate the ability of the quadrupole system to facilitate remote nsPEF electropermeabilization. Numerical simulations were employed to approximate the nsPEF distribution for a two-dimensional (2-D) area resulting from unipolar, bipolar or CANCAN exposure in a varied-pulse quadrupole electrode configuration. For all experiments, the independent variables were fixed for pulse width (600 ns), pulse number (50) and repetition rate (10 Hz). Electropermeabilization served as the biological endpoint, with green fluorescence due to cell uptake of the nuclear dye YO-PRO-1® (YP1) tracer molecule serving the response variable. An agarose-based 3-D tissue model was used to acquire, quantify and compare fluorescence intensity data in vitro, which was measured by stereomicroscopy to enable macro versus micro level 2-D visualization.

Results of this investigation showed that increasing the magnitude of the applied voltage shifts unipolar responses from localization at the anodal to cathodal electrode, and that adding a second proximal ground electrode increases the response area. Bipolar nsPEF responses were generally less intense than unipolar, but these depended on both the inter-electrode location measured and amplitude of the second phase. CANCAN preliminary indicated some ability to decrease strong uptake at electrodes, but evaluation across experimental and published data indicate that greater differences between unipolar and bipolar responses are needed to improve possibilities for distal stimulation. Overall, this work demonstrated the potential for more complex pulser-electrode configurations to successfully modulate nsPEF electropermeabilization in space by controlling unipolar and bipolar pulse delivery and contributed to a deeper understanding of bipolar cancellation. By providing a set of metrics for test and evaluation, the data provided herein may serve to inform model development to support prediction of nsPEF outcomes and help to more acutely define spatial-intensity relationships between nsPEFs and cell permeabilization as well as delineate requirements for future non-invasive nsPEF therapies.

DOI

10.25777/89kj-z744

ISBN

9798678109828

ORCID

0000-0002-2373-3739

Available for download on Wednesday, March 31, 2021

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