Effect of Experimental Electrical and Biological Parameters on Gene Transfer by Electroporation: A Systematic Review and Meta-Analysis.

The exact mechanisms of nucleic acid (NA) delivery with gene electrotransfer (GET) are still unknown, which represents a limitation for its broader use. Further, not knowing the effects that different experimental electrical and biological parameters have on GET additionally hinders GET optimization, resulting in the majority of research being performed using a trial-and-error approach. To explore the current state of knowledge, we conducted a systematic literature review of GET papers in in vitro conditions and performed meta-analyses of the reported GET efficiency. For now, there is no universal GET strategy that would be appropriate for all experimental aims. Apart from the availability of the required electroporation device and electrodes, the choice of an optimal GET approach depends on parameters such as the electroporation medium; type and origin of cells; and the size, concentration, promoter, and type of the NA to be transfected. Equally important are appropriate controls and the measurement or evaluation of the output pulses to allow a fair and unbiased evaluation of the experimental results. Since many experimental electrical and biological parameters can affect GET, it is important that all used parameters are adequately reported to enable the comparison of results, as well as potentially faster and more efficient experiment planning and optimization.

Early Cost-effectiveness Analysis of Electrochemotherapy as a Prospect Treatment Modality for Skin Melanoma.

PURPOSE: Electrochemotherapy is increasingly entering into national and international guidelines, requiring formal evaluation of treatment costs and cost-effectiveness to ensure that its uptake provides value to budget-constrained health care systems. This study analyzed the early cost-effectiveness of electrochemotherapy in patients with Stage IIIc/IV skin melanoma in clinical practice in Slovenia. The costs of electrochemotherapy were compared to those of the standard of care, consisting of palliative treatment and therapy for symptoms. METHODS: wThe study enrolled 23 patients treated with electrochemotherapy at the Institute of Oncology (Ljubljana, Slovenia). The mean cost of electrochemotherapy was estimated using patient-specific cost data on electrochemotherapy procedures and subsequent follow-up. Quality-adjusted life-years (QALYs) were estimated by collecting EQ-5D-3L questionnaires at baseline, after complete or partial response following the treatment, and after a relapse of skin lesions. A discrete-time Markov model was built to estimate the lifetime costs and consequences of using electrochemotherapy compared to standard of care, from the perspective of the Slovenian health care system. The analysis was conducted separately in the whole patient sample and in the subset of patients with bleeding lesions. Deterministic and probabilistic sensitivity analyses were conducted to test model assumptions and to characterize the uncertainty around model parameters. FINDINGS: In the whole patient population, electrochemotherapy for skin melanoma Stage IIIc/IV was expected to increase QALYs by 0.29 (95% credible interval [CrI], 0.10-0.50), at the higher cost of 6568 EUR (95% CrI, 4593-8928) in comparison to the standard of care. At the cost-effectiveness threshold of 20,000 EUR/QALY, the estimated probabilities of electrochemotherapy being cost-effective compared to standard of care were 0.30 and 0.91 in the whole patient sample and in patients with bleeding lesions, respectively. In the whole sample population, a 50% reduction in the price of the electrodes was expected to increase the probability of electrochemotherapy being cost-effective from 0.30 to ~0.64. IMPLICATIONS: The findings from this cost-effectiveness analysis of data from clinical practice were based on a small sample size (ie, 23 patents), which made the subgroup of patients with bleeding lesions very small. Therefore, the findings in this patient population should be carefully interpreted.

Mechanistic view of skin electroporation - models and dosimetry for successful applications: an expert review.

Introduction: Skin electroporation is a promising treatment for transdermal drug delivery, gene electrotransfer, skin rejuvenation, electrochemotherapy, and wound disinfection. Although a considerable amount of in vitro and in vivo studies exists, the translation to clinics is not as fast as one would hope. We hypothesize the reason lies in the inadequate dosimetry, i.e. electrode configurations, pulse parameters, and pulse generators used. We suggest adequate dosimetry can be determined by mathematical modeling which would allow comparison of protocols and facilitate translation into clinics.Areas covered: We introduce the mechanisms and applications of skin electroporation, present existing mathematical models and compare the influence of different model parameters. We review electrodes and pulse generators, prototypes, as well as commercially available models.Expert opinion: The reasons for slow translation of skin electroporation treatments into clinics lie in uncontrolled and inadequate dosimetry, poor reporting rendering comparisons between studies difficult, and significant differences in animal and human skin morphology often dismissed in reports. Mathematical models enable comparison of studies, however, when the parameters of the pulses and electrode configuration are not adequately reported, as is often the case, comparisons are difficult, if not impossible. For each skin electroporation treatment, systematic studies determining optimal parameters should be performed and treatment parameters standardized.

Nanosecond Pulse Electroporator With Silicon Carbide mosfets: Development and Evaluation.

Nanosecond electroporation of cell organelles is being studied since more than a decade, but it is still not entirely understood. Unique prototype hardware equipment and challenging measuring methods may also be a contributing reason for this situation. In the scope of this paper, we improve the performance of the high-voltage nanosecond pulse generator by introducing silicon carbide (SiC) mosfets. We developed a new high-voltage diode opening switch (DOS)-nanosecond pulse generator for laboratory use for in vitro experiments in electroporation cuvettes. Analysis and comparison of the most commonly used switching technologies in pulse generators were made. The device is designed by two parallel two-stage Marx-bank circuits with SiC mosfets that generates up to 200 A in the resonant network. A driving circuit for stable simultaneous switching of SiC mosfets was developed. The developed generator can deliver from 500 V to more than 6 kV, approximately 8 ns pulses to a 50 Ω load. Even though the amplitude of the output pulse is not as high as expected, the multiplication factor [Formula: see text] is still approximately 9, which is an improvement compared to the previously published linear DOS generator. Measurement and evaluation process is described in detail. Additionally, we emphasize on the size of an error that occurs during measurements.