Publications by authors named "Mark J Jaroszeski"

17 Publications

  • Page 1 of 1

Real-time impedance feedback to enhance cutaneous gene electrotransfer in a murine skin model.

Bioelectrochemistry 2021 Jul 13;142:107885. Epub 2021 Jul 13.

Department of Chemical, Biological, and Materials Engineering, University of South Florida, 4202 E. Fowler Ave ENG 030, Tampa, FL 33620, USA; Center for Molecular Delivery at USF, University of South Florida, 4202 E. Fowler Ave ENG 030, Tampa, FL 33620, USA; Department of Medical Engineering, University of South Florida, 4202 E. Fowler Avenue ENG 030, Tampa, FL 33620, USA. Electronic address:

Electric field mediated gene delivery methods have the ability to efficiently transfect cells in vivo with an excellent safety profile. The method has historically used a fixed number of electric pulses with identical characteristics in induce delivery. Electrical treatment does not typically compensate for subject-to-subject variation and other differences. This study was designed to investigate if delivery/expression could be increased using a novel electropulsation method that compensated for variation using real-time electrical impedance measurements. The method involved delivering plasmid DNA encoding luciferase to murine skin. Tissue impedance in a 1-3 KHz range was measured before electric pulses were applied. Impedance was also measured after each successive pulse. Pulsation was stopped when impedance values were reduced by either 80% or 95% relative to prepulse values. Standard/fixed pulsing parameters were also used for comparison. The results indicated that up to 15-fold increases in luciferase expression could be obtained when electrical treatment was ceased based upon impedance reductions. Furthermore, peak expression levels of all treatment groups pulsed using the novel pulsing method were statistically higher than those that employed standard pulsing. These results strongly suggest that applying pulses until a defined impedance-based endpoint results in higher expression.
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http://dx.doi.org/10.1016/j.bioelechem.2021.107885DOI Listing
July 2021

Impedance spectroscopy as an indicator for successful in vivo electric field mediated gene delivery in a murine model.

Bioelectrochemistry 2017 Jun 27;115:33-40. Epub 2017 Jan 27.

Department of Chemical and Biomedical Engineering, University of South Florida, 4202 E. Fowler Ave. ENB 118, Tampa, FL 33620, USA; Center for Molecular Delivery at USF, University of South Florida, 4202 E. Fowler Ave. ENB 118, Tampa, FL 33620, USA.

In vivo gene electro transfer technology has been very successful both in animal models and in clinical trials over the past 20years. However, variable transfection efficiencies can produce inconsistent outcomes. This can be due to differences in tissue architecture and/or chemical composition which may effectively create unique biological environments from subject to subject that may respond differently to the identical electric pulses. This study investigates the integration of impedance spectroscopy into the gene electro transfer process to measure murine skin impedance spectra before, during (after pulse delivery), and after gene electro transfer pulse application to determine if changes in impedance correlate with reporter gene expression. Both post-treatment impedance spectra and gene expression were dependent upon the applied electric field strength. These results indicate that alterations in tissue impedance produced by the applied electric field represent an excellent parameter to predict degrees of transfection and gene expression. These results could ultimately be used to alter pulsing parameters in order to optimize delivery/expression.
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http://dx.doi.org/10.1016/j.bioelechem.2017.01.004DOI Listing
June 2017

Direct Current Helium Plasma for Delivery of Plasmid DNA Encoding Erythropoietin to Murine Skin.

Plasma Med 2017 ;7(3):261-271

Dept of Chemical and Biomedical Engineering, University of South Florida, College of Engineering, Tampa, FL.

The use of electric fields to deliver DNA, called electroporation, has the potential to broadly impact vaccination and disease treatment. The evidence for this has emerged from a large number of recently completed and ongoing clinical trials. The methods for applying electric fields to tissues traditionally involve contact between metal electrodes and the tissue. In this study, we investigated the use of helium plasma as a noncontact method for electrically treating tissue in a manner that results in the uptake and expression of foreign DNA in murine skin. More specifically, our goal was to demonstrate that DNA encoding a model-secreted protein could be delivered, detected in the blood, and remain functional to produce its known biological effect. Murine erythropoietin (EPO) was the model-secreted protein. Results clearly demonstrated that an intradermal DNA injection followed by plasma treatment for 2 min resulted in elevated levels of EPO in the blood and corresponding hemoglobin increases that were statistically significant relative to DNA injection alone.
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http://dx.doi.org/10.1615/PlasmaMed.2017019506DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6404545PMC
January 2017

Fluorometric assay to compensate for non-viable cells during electroporation.

J Fluoresc 2015 Jan 11;25(1):159-65. Epub 2014 Dec 11.

Department of Chemical and Biomedical Engineering, College of Engineering, University of South Florida, ENB 118, 4202 E. Fowler Ave, Tampa, FL, 33620, USA.

A fluorometric assay is described that allows adjustment for non-viable cells that result during electroporation. The technique, unlike others, relies on only one dye, requires a single instrument, and eliminates the need for a separate cell counting step. Murine melanoma (B16-F10) cells were electroporated using electric fields ranging from 400 to 2500 V/cm in the presence of SYTOX(®)-green. Compensation for the fluorescence resulting from non-viable cells was facilitated by a correction curve established by lysing a known number of cells in the presence of SYTOX(®)-green. In uncorrected data, an applied electric field of 2500 V/cm increased dye delivery but also reduced cell viability significantly. Compensating for the fluorescence of non-viable cells showed that changing the field strength to 800 V/cm or 2500 V/cm from 400 V/cm had only marginal effects on membrane pore formation. The fluorometric assay was used to compare electroporation in high conductivity (PBS) and a low conductivity medium (LC-PBS). Statistically significant increases of 10 to 30-fold were observed for cells electroporated at 400 V/cm and 800 V/cm in LC-PBS.
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http://dx.doi.org/10.1007/s10895-014-1492-yDOI Listing
January 2015

Optimization of a plasma facilitated DNA delivery method.

Bioelectrochemistry 2015 Jun 13;103:15-21. Epub 2014 Oct 13.

Department of Chemical and Biomedical Engineering, College of Engineering, University of South Florida, 4202 East Fowler Avenue, Tampa, FL 33620, United States. Electronic address:

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.
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http://dx.doi.org/10.1016/j.bioelechem.2014.09.003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4346600PMC
June 2015

Electrogenetherapy of B16.F10 murine melanoma tumors with an interleukin-28 expressing DNA plasmid.

Hum Vaccin Immunother 2012 Nov 1;8(11):1722-8. Epub 2012 Nov 1.

Department of Molecular Medicine; Morsani College of Medicine; Tampa, FL USA.

Augmented delivery of cytokine-expressing DNA plasmids to subcutaneous tumors has been demonstrated to result in a level of enhanced anti-tumor activity. One delivery enhancement method which has been evaluated is in vivo electroporation (EP), a contact-dependent delivery technique where electric pulses are hypothesized to augment the transfer of DNA into cells and tissues through the induction of temporary cell membrane pores. Previous work by members of our group, as well as others, has demonstrated the anti-tumor effects of DNA plasmids expressing the cytokines IL-12 and IL-15. In this report the potential anti-tumor activity of a relatively newly-described cytokine, IL-28, was measured when administered intratumorally as a DNA expression plasmid (designated pIL28) to established murine (B16.F10) melanoma tumors. The administration of the IL-28 expressing plasmid was performed through enhanced delivery methods. One method was EP and the other a non-contact dependent technique using a helium plasma stream. IL-28 is a member of the type III interferon family of cytokines that has been characterized as possessing potent anti-viral activity. This cytokine has been demonstrated to function as an adjuvant in small animal model vaccination protocols and stimulates CD8+ CTL responses. In addition, stimulation of anti-tumor activity has been demonstrated in several studies using IL-28. Based on these activities, it was hypothesized that this cytokine could, when delivered through a DNA expression plasmid, mediate anti-tumor activity. The results of this study indicated that enhanced delivery of pIL-28 resulted in attenuation of tumor growth, compared with non-enhanced delivery. Of note, this is the first proof-of-concept experiment, of our knowledge, documenting the ability of a non-contact dependent helium plasma-based delivery method to mediate the enhancement of an anti-tumor effect by a cytokine-expressing DNA plasmid. This suggests the use of the helium plasma delivery method as an alternative or adjunctive method to EP for the effective delivery of agents that possess potential anti-tumor activity.
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http://dx.doi.org/10.4161/hv.22560DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3601148PMC
November 2012

Non-contact helium-based plasma for delivery of DNA vaccines. Enhancement of humoral and cellular immune responses.

Hum Vaccin Immunother 2012 Nov 16;8(11):1729-33. Epub 2012 Aug 16.

Center for Molecular Delivery, University of South Florida; Tampa, FL, USA.

Non-viral in vivo administration of plasmid DNA for vaccines and immunotherapeutics has been hampered by inefficient delivery. Methods to enhance delivery such as in vivo electroporation (EP) have demonstrated effectiveness in circumventing this difficulty. However, the contact-dependent nature of EP has resulting side effects in animals and humans. Noncontact delivery methods should, in principle, overcome some of these obstacles. This report describes a helium plasma-based delivery system that enhanced humoral and cellular antigen-specific immune responses in mice against an intradermally administered HIV gp120-expressing plasmid vaccine (pJRFLgp120). The most efficient plasma delivery parameters investigated resulted in the generation of geometric mean antibody-binding titers that were 19-fold higher than plasmid delivery alone. Plasma mediated delivery of pJRFLgp120 also resulted in a 17-fold increase in the number of interferon-gamma spot-forming cells, a measure of CD8+ cytotoxic T cells, compared with non-facilitated plasmid delivery. This is the first report demonstrating the ability of this contact-independent delivery method to enhance antigen-specific immune responses against a protein generated by a DNA vaccine.
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http://dx.doi.org/10.4161/hv.21624DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3601149PMC
November 2012

Enhancement of antigen specific humoral immune responses after delivery of a DNA plasmid based vaccine through a contact-independent helium plasma.

Vaccine 2011 Sep 31;29(39):6781-4. Epub 2010 Dec 31.

Department of Chemical and Biomedical Engineering, College of Engineering, University of South Florida, 4202 E. Fowler Avenue, ENB 118, Tampa, FL 33620, USA.

Non-viral in vivo delivery of DNA, encoding for specific proteins, has traditionally relied on chemical or physical forces applied directly to tissues. Physical methods typically involve contact between an applicator/electrode and tissue and often results in transient subject discomfort. To overcome these limitations of contact-dependent delivery, a helium plasma source was utilized to deposit ionized gasses to treatment/vaccination sites without direct contact between the applicator and the tissues. The study reported here evaluated the efficacy of this strategy as an effective method to administer DNA vaccines. Balb/C mice were vaccinated with a DNA plasmid expressing an HIVgp120 envelope glycoprotein either with or without co-administration of helium plasma or electroporation. The results indicated, for the first time, the potential efficacy of helium plasma delivery for the induction and enhancement of antigen specific immune responses following DNA vaccination.
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http://dx.doi.org/10.1016/j.vaccine.2010.12.054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3156891PMC
September 2011

Characterization of plasma mediated molecular delivery to cells in vitro.

Int J Pharm 2010 Apr 18;389(1-2):53-7. Epub 2010 Jan 18.

Department of Chemical and Biomedical Engineering, University of South Florida, ENB 118, 4202 E. Fowler Avenue, Tampa, FL 33620, USA.

Ion-based strategies have recently emerged as a method to facilitate molecular delivery. These methods are attractive as they separate the applicator from the treatment site avoiding some issues encountered with other electrically driven methods. Current literature on plasma delivery has shown utility in vitro and in vivo for both drugs and genes. To advance this technology more information must become available on the mechanism responsible for delivery and the effects of ion exposure on eukaryotic cells. This in vitro investigation found that molecular delivery facilitated by a DC-based plasma follows a dose-response behavior, with optimum uptake of Sytox Green occurring in two cell lines after 600 s of exposure. In both cell lines exposure to the discharge caused no adverse effects in viability for exposure times up to 600 s. It was also found that membranes treated with ions remained permeabilized for several minutes following plasma treatment and that membrane resealing exhibited first order kinetics.
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http://dx.doi.org/10.1016/j.ijpharm.2010.01.016DOI Listing
April 2010

Electrically mediated delivery of plasmid DNA to the skin, using a multielectrode array.

Hum Gene Ther 2010 Mar;21(3):357-62

Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA.

The easy accessibility of skin makes it an excellent target for gene transfer protocols. To take full advantage of skin as a target for gene transfer, it is important to establish an efficient and reproducible delivery system. Electroporation is a strong candidate to meet this delivery criterion. Electroporation of the skin is a simple, direct, in vivo method to deliver genes for therapy. Previously, delivery to the skin was performed by means of applicators with relatively large distances between electrodes, resulting in significant muscle stimulation and pain. These applicators also had limitations in controlling the directionality of the applied field. To resolve this issue, a system consisting of an array of electrodes that decreased the distance between them and that were independently addressable for directional control of the field was developed. This new multielectrode array (MEA) was compared with an established electrode. In a rat model, comparable reporter expression was seen after delivery with each electrode. Delivery was also evaluated in a guinea pig model to determine the potential of this approach in an animal model with skin thickness and structure similar to human skin. The results clearly showed that effective delivery was related to both the electrode and the parameters chosen. With the MEA, the muscle twitching associated with application of electric fields was notably reduced compared with conventional electrode systems. This is important, as it will facilitate the translation of electroporation-mediated gene delivery to skin for clinical use with DNA vaccines or for therapies for cancer or protein deficiencies.
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http://dx.doi.org/10.1089/hum.2009.065DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2865213PMC
March 2010

Plasma facilitated delivery of DNA to skin.

Biotechnol Bioeng 2009 Dec;104(5):1034-40

Department of Chemical and Biomedical Engineering, University of South Florida, ENB 118, Tampa, Florida 33620, USA.

Non-viral delivery of cell-impermeant drugs and DNA in vivo has traditionally relied upon either chemical or physical stress applied directly to target tissues. Physical methods typically use contact between an applicator, or electrode, and the target tissue and may involve patient discomfort. To overcome contact-dependent limitations of such delivery methodologies, an atmospheric helium plasma source was developed to deposit plasma products onto localized treatment sites. Experiments performed in murine skin showed that samples injected with plasmid DNA encoding luciferase and treated with plasma demonstrated increased levels of expression relative to skin samples that received injections of DNA alone. Increased response relative to injection alone was observed when either positive or negative voltage was used to generate the helium plasma. Quantitative results over a 26-day follow-up period showed that luciferase levels as high as 19-fold greater than the levels obtained by DNA injection alone could be achieved. These findings indicate that plasmas may compete with other physical delivery methodologies when skin is the target tissue.
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http://dx.doi.org/10.1002/bit.22451DOI Listing
December 2009

Comparison of electrically mediated and liposome-complexed plasmid DNA delivery to the skin.

Genet Vaccines Ther 2008 Dec 4;6:16. Epub 2008 Dec 4.

Center for Molecular Delivery, University of South Florida, Tampa, FL, USA.

Background: Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery.

Methods: Enhanced electrically-mediated delivery, and less extensively, liposome complexed delivery, of a plasmid encoding the reporter luciferase was tested in rodent skin. Expression kinetics and tissue damage were explored as well as testing in a second rodent model.

Results: Experiments confirm that electroporation alone is more effective in enhancing reporter gene expression than plasmid injection alone, plasmid conjugation with liposomes followed by injection, or than the combination of liposomes and electroporation. However, with two time courses of multiple electrically-mediated plasmid deliveries, neither the levels nor duration of transgene expression are significantly increased. Tissue damage may increase following a second treatment, no further damage is observed after a third treatment. When electroporation conditions utilized in a mouse model are tested in thicker rat skin, only higher field strengths or longer pulses were as effective in plasmid delivery.

Conclusion: Electroporation enhances reporter plasmid delivery to the skin to a greater extent than the liposome conjugation method tested. Multiple deliveries do not necessarily result in higher or longer term expression. In addition, some impact on tissue integrity with respect to surface damage is observed. Pulsing conditions should be optimized for the model and for the expression profile desired.
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http://dx.doi.org/10.1186/1479-0556-6-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2631522PMC
December 2008

Feasibility study for focusing electric fields to mediate in vitro drug and gene delivery.

Conf Proc IEEE Eng Med Biol Soc 2006;2006:5617-20

Department of Chemical Engineering, University of South Florida, Tampa, FL 33620, USA.

Electric field mediated drug and gene delivery is a novel method that uses pulsed electric fields to improve permeability of cell membranes and therefore desired agent uptake by tissues. In this paper, we describe the modeling and experimental proof of concept of a method to direct electric fields to subsequently focus drug or gene uptake at a desired site. The in vitro experimental results presented are consistent with simulation models and could be scaled into different in vivo applications that can concentrate the effects of electroporation and overcome several problems related to localized effects near the electrodes.
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http://dx.doi.org/10.1109/IEMBS.2006.259865DOI Listing
March 2008

Electrically mediated plasmid DNA delivery to solid tumors in vivo.

Methods Mol Biol 2004 ;245:237-44

Department of Chemical Engineering, College of Engineering, University of South Florida, Tampa, FL, USA.

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http://dx.doi.org/10.1385/1-59259-649-5:237DOI Listing
February 2004

Electrochemotherapy for the treatment of human sarcoma in athymic rats.

Technol Cancer Res Treat 2002 Oct;1(5):393-9

Department of Chemical Engineering, College of Engineering, University of South Florida, Tampa, FL 33612, USA.

Electrochemotherapy is the combined use of a chemotherapeutic agent and pulsed electric fields. Electrical treatment causes an increase in cell membrane permeability which allows the chemotherapeutic agent to more freely enter the tumor cells. Electrochemotherapy has been under development in clinical trials. This study focused on determining the applicability of electrochemotherapy for treating soft tissue sarcoma using an animal model bearing human sarcomas. The antitumor effects of several concentrations of cisplatin, bleomycin, doxorubicin, and netropsin as single agents delivered with electric pulses were investigated based on post-treatment tumor volumes and histology. Electrochemotherapy treatment resulted in 5% to 88.9% durable complete responses; ECT that employed bleomycin resulted in the highest antitumor effects. This indicates the feasibility of electrochemotherapy as a modality for limb preserving treatments for sarcoma of the extremities.
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http://dx.doi.org/10.1177/153303460200100510DOI Listing
October 2002

Effect of electrochemotherapy on muscle and skin.

Technol Cancer Res Treat 2002 Oct;1(5):385-92

Center for Molecular Delivery, University of South Florida, Tampa, FL 33612, USA.

The efficient delivery of drugs to tumors is an important tool for the treatment of a variety of cancers. Electric pulses have been shown to facilitate the uptake of molecules through the cell membrane. This procedure has been successful in increasing the effectiveness of anti-tumor agents (electrochemotherapy; ECT). Response rates of >80% have been obtained in both animal and human trials for several types of skin malignancies. The study reported here examined the effect of ECT on normal tissue. The hind limbs of Sprague Dawley rats were treated with 1-3 electroporation sequences in the presence or absence of the drug (bleomycin) which was administered at 4, 8 or 16 units/ml. The treated sites were examined histologically 3, 14 and 56 days later. Limb function was not affected by the treatment and skin and muscle necrosis was only seen at the higher doses.
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http://dx.doi.org/10.1177/153303460200100509DOI Listing
October 2002

Electric field enhanced plasmid delivery to liver hepatocellular carcinomas.

Technol Cancer Res Treat 2002 Oct;1(5):355-64

Center for Molecular Delivery, University of South Florida, USA.

Electric field enhanced molecular delivery for cancer research and treatment is a new technology that has demonstrated its effectiveness in clinical trials using bleomycin or cisplatin (Heller, R., Gilbert, R., Jaroszeski, M. J. Clinical applications of electrochemotherapy, Advanced Drug Delivery Reviews 35,119-129, 1999), as chemotherapeutic agents. The technology is being investigated in research applications for applicability as a method to enhance gene expression in a target tumor. Success is predicated on an appropriate effective electric field mediated delivery protocol that triggers significant appropriate gene expression duration and levels. An electric field mediated delivery protocol includes a set of conditions associated with the electric field, the electroporation signature, as well as parameters associated with the plasmid and the electric field applicator. Manipulation of the electrical parameters within the electroporation signature generates different gene expression levels in liver hepatocellular carcinomas. Statistically significant gene expression levels were obtained that differed by an order of magnitude when two different electric field strength and duration conditions were employed.
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http://dx.doi.org/10.1177/153303460200100506DOI Listing
October 2002
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