title: Optimization of a plasma facilitated DNA delivery method.
abstract: Plasma-based methods have recently emerged as a technique for augmenting plasmid DNA delivery to skin. This delivery modality relies on the deposition of ionized gas molecules on to targeted cells or tissue to establish an electric field. It is hypothesized that this electric field results in the dielectric breakdown of cell membranes, making cells permeable to exogenous molecules. This in vivo investigation sought to optimize the intradermal delivery of a luciferase expressing plasmid DNA by modulating the total exposure to the plasma source and the plasmid DNA dose. Varying the plasma exposure time from 2, 5, 10, and 20 min allowed the conditions resulting in the highest expression of luciferase to be found. These conditions correlated to the 10 minute exposure time for a plasma derived from either +8 kV or -8 kV, when the generator was operated 3 cm from the epidermal tissue surface with a helium flow rate of 15 L/min. Exposing the injected flank skin for 10 min resulted in a rise of 37.3-fold for a plasma created with +8 kV and 27.1-fold for a plasma created with -8 kV. When using this treatment time with 50, 100, or 200 μg of a luciferase expressing plasmid, it was found that 100 μg resulted in the highest peak luminescence.
authors.affiliation: Department of Chemical and Biomedical Engineering, College of Engineering, University of South Florida, 4202 East Fowler Avenue, Tampa, FL 33620, United States.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Connolly
authors.affiliation: Department of Electrical Engineering, College of Engineering, University of South Florida, 4202 East Fowler Avenue, Tampa, FL 33620, United States.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Hoff
authors.affiliation: Department of Chemical and Biomedical Engineering, College of Engineering, University of South Florida, 4202 East Fowler Avenue, Tampa, FL 33620, United States.
authors.forename: Richard
authors.initials: R
authors.lastname: Gilbert
authors.affiliation: Department of Chemical and Biomedical Engineering, College of Engineering, University of South Florida, 4202 East Fowler Avenue, Tampa, FL 33620, United States. Electronic address: mjarosze@usf.edu.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Jaroszeski
mesh_terms: D000818:Animals; D004247:DNA / Q000008:administration & dosage*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D018274:Electroporation / Q000379:methods; D004867:Equipment Design; D005260:Female; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D008810:Mice, Inbred C57BL; D058626:Plasma Gases; D010957:Plasmids / Q000008:administration & dosage; D012867:Skin; D013997:Time Factors
chemical_list: D058626:Plasma Gases; D004247:DNA; D008156:Luciferases
keywords: Electroporation; Molecular delivery; Plasma; Plasma delivery; Plasmid DNA; Skin