Date of Award

Spring 2015

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Program/Concentration

Biology

Committee Director

Richard Heler

Committee Director

Hameeda Sultana

Committee Member

Robert R. Ratzlaff

Call Number for Print

Special Collections LD4331.B46 E245 2015

Abstract

Cold plasma is produced when strong applied electric fields accelerate free electrons, which dissociate, excite, or ionize gaseous molecules [1]. The deposition of ions from the plasma source is dependent on power generation, input gas composition, and gas flow rate. In the presence of reactive species, the membrane of eukaryotic cells is compromised allowing for otherwise impermeant molecules, such as DNA, to enter the inner-cell milieu [2].

The efficacy of a novel cold plasma reactor based on shielded sliding discharge for the delivery of plasmid DNA was assessed. The device is entirely non-contact, wherein the plasma never directly touches the target surface. Instead, the plasma-activated air (PAA) is dispersed on-site. Experiments were performed with mouse melanoma cells (B16F10) and human keratinocyte cells (HaCaT) inoculated with plasmid DNA encoding luciferase (pLUC). Quantitative results measured over a 72-hour period displayed luciferase expression levels as high as 3-fold greater in cells exposed to PAA than levels obtained from the inoculation of plasmid DNA alone (p

Delivery of plasmid encoding GFP (pGFP) to HaCaT cells seeded on PCL scaffolds was confirmed by immunostaining. In addition, reporter genes were delivered to recellularized DermACELL® human skin constructs using PAA. Delivery of pLUC was enhanced with PAA, resulting in a 4-fold increase in luciferase expression over 120 hours compared to the injection of pLUC alone (p

In vivo experiments were performed in C57BL/6 and BALB/c mice, with skin as the delivery target. PAA exposure significantly enhanced luciferase expression in C57BL/6 mice over 28 days compared to intradermal pLUC injection alone (p

Cold plasma for DNA delivery is attractive as it provides a non-viral, noninvasive method. Our findings suggest PAA-mediated delivery warrants further exploration as an alternative or supplemental approach for DNA transfer.

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DOI

10.25777/dzpk-ht92

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