Negative effects of cancellation during nanosecond range High-Frequency calcium based electrochemotherapy in vitro

Rembiałkowska, Nina; Szlasa, Wojciech; Radzevičiūtė-Valčiukė, Eivina; Kulbacka, Julita; Novickij, Vitalij

doi:10.1016/j.ijpharm.2023.123611

International Journal of Pharmaceutics

2023

DOI: 10.1016/j.ijpharm.2023.123611

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Negative effects of cancellation during nanosecond range High-Frequency calcium based electrochemotherapy in vitro

Nina Rembiałkowska,

Wojciech Szlasa,

Eivina Radzevičiūtė-Valčiukė

et al.

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2024

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Cited by 3 publications

References 63 publications

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Enhancing lung cancer growth inhibition with calcium ions: Role of mid- and high-frequency electric field pulses

Rembiałkowska,

Kucharczyk,

Radzevičiūtė-Valčiukė

et al. 2024

Biomedicine & Pharmacotherapy

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Enhancing lung cancer growth inhibition with calcium ions: Role of mid- and high-frequency electric field pulses

Rembiałkowska,

Kucharczyk,

Radzevičiūtė-Valčiukė

et al. 2024

Biomedicine & Pharmacotherapy

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Threshold Interphase Delay for Bipolar Pulses to Prevent Cancellation Phenomenon during Electrochemotherapy

Malyško-Ptašinskė,

Nemeikaitė-Čėnienė,

Radzevičiūtė-Valčiukė

et al. 2024

IJMS

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Electroporation-based procedures employing nanosecond bipolar pulses are commonly linked to an undesirable phenomenon known as the cancelation effect. The cancellation effect arises when the second pulse partially or completely neutralizes the effects of the first pulse, simultaneously diminishing cells’ plasma membrane permeabilization and the overall efficiency of the procedure. Introducing a temporal gap between the positive and negative phases of the bipolar pulses during electroporation procedures may help to overcome the cancellation phenomenon; however, the exact thresholds are not yet known. Therefore, in this work, we have tested the influence of different interphase delay values (from 0 ms to 95 ms) using symmetric bipolar nanoseconds (300 and 500 ns) on cell permeabilization using 10 Hz, 100 Hz, and 1 kHz protocols. As a model mouse hepatoma, the MH-22a cell line was employed. Additionally, we conducted in vitro electrochemotherapy with cisplatin, employing reduced interphase delay values (0 ms and 0.1 ms) at 10 Hz. Cell plasma membrane permeabilization and viability dependence on a variety of bipolar pulsed electric field protocols were characterized. It was shown that it is possible to minimize bipolar cancellation, enabling treatment efficiency comparable to monophasic pulses with identical parameters. At the same time, it was highlighted that bipolar cancellation has a significant influence on permeabilization, while the effects on the outcome of electrochemotherapy are minimal.

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The Effects of Bipolar Cancellation Phenomenon on Nano-Electrochemotherapy of Melanoma Tumors: In Vitro and In Vivo Pilot

Mickevičiūtė,

Radzevičiūtė-Valčiukė,

Malyško-Ptašinskė

et al. 2024

IJMS

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The phenomenon known as bipolar cancellation is observed when biphasic nanosecond electric field pulses are used, which results in reduced electroporation efficiency when compared to unipolar pulses of the same parameters. Basically, the negative phase of the bipolar pulse diminishes the effect of the positive phase. Our study aimed to investigate how bipolar cancellation affects Ca2+ electrochemotherapy and cellular response under varying electric field intensities and pulse durations (3–7 kV/cm, 100, 300, and 500 ns bipolar 1 MHz repetition frequency pulse bursts, n = 100). As a reference, standard microsecond range parametric protocols were used (100 µs × 8 pulses). We have shown that the cancellation effect is extremely strong when the pulses are closely spaced (1 MHz frequency), which results in a lack of cell membrane permeabilization and consequent failure of electrochemotherapy in vitro. To validate the observations, we have performed a pilot in vivo study where we compared the efficacy of monophasic (5 kV/cm × ↑500 ns × 100) and biphasic sequences (5 kV/cm × ↑500 ns + ↓500 ns × 100) delivered at 1 MHz frequency in the context of Ca2+ electrochemotherapy (B16-F10 cell line, C57BL/6 mice, n = 24). Mice treated with bipolar pulses did not exhibit prolonged survival when compared to the untreated control (tumor-bearing mice); therefore, the bipolar cancellation phenomenon was also occurrent in vivo, significantly impairing electrochemotherapy. At the same time, the efficacy of monophasic nanosecond pulses was comparable to 1.4 kV/cm × 100 µs × 8 pulses sequence, resulting in tumor reduction following the treatment and prolonged survival of the animals.

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Negative effects of cancellation during nanosecond range High-Frequency calcium based electrochemotherapy in vitro

Cited by 3 publications

References 63 publications

Enhancing lung cancer growth inhibition with calcium ions: Role of mid- and high-frequency electric field pulses

Enhancing lung cancer growth inhibition with calcium ions: Role of mid- and high-frequency electric field pulses

Threshold Interphase Delay for Bipolar Pulses to Prevent Cancellation Phenomenon during Electrochemotherapy

The Effects of Bipolar Cancellation Phenomenon on Nano-Electrochemotherapy of Melanoma Tumors: In Vitro and In Vivo Pilot

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