Effectiveness Of Electrochemotherapy Research Articles

Treatment benefit of electrochemotherapy for superficial squamous cell carcinoma: a systematic review and single-arm meta-analysis.

Treating aggressive superficial squamous cell carcinoma (SCC) poses challenges due to invasiveness. Palliative care is recommended for inoperable cases with extensive tumors near vital organs, risking disfigurement or functional impairment. Electrochemotherapy (ECT) is an emerging cutaneous tumor treatment, but its efficacy against superficial SCC remains uncertain. This study conducts a systematic review and single-arm meta-analysis to evaluate ECT's effectiveness against superficial SCC and provide current evidence for clinical practice. Embase, PubMed and Cochrane Library were searched for studies up to May 2023. The random effects model analyzed complete response (CR) and partial response (PR), with subgroup assessment based on drug dosage, treatment response evaluation, tumor size, primary/recurrent status, and tumor location. Ten studies involving 162 patients and 208 tumors were included. Pooled CR and PR rates for ECT-treated superficial SCC were 66.5% (95% CI 48.4%-82.5%; I2 = 84%) and 20.3% (95% CI 10.5%-32.3%; I2 = 70%), respectively. Subgroup analysis indicated ECT's superiority in treating primary tumors (PR: 70%, CR: 30%) and tumors ≤ 3cm (PR: 81.3%, CR: 10.1%) compared to recurrent tumors (PR: 56.7%, CR: 36.5%) and tumors > 3cm (PR: 45.2%, CR: 34.4%). This single-arm meta-analysis confirms ECT's efficacy against superficial SCC, especially in primary tumors and those ≤ 3cm in diameter. The study highlights the impact of tumor location and response evaluation on ECT's benefits, warranting further investigation through additional research.

Effect of electrochemotherapy on myogenesis of mouse C2C12 cells in vitro

Electrochemotherapy (ECT) is a local ablative therapy for the treatment of different skin and subcutaneous tumors and certain tumors in internal organs. Skeletal muscle represents a major tumor- surrounding tissue, exposed to side effects of ECT. At the cellular level, side-effects of ECT on skeletal muscle and underlying mechanisms have not been examined yet. Thus, we aimed to determine the effect of ECT in the mouse muscle cell line C2C12 during in vitro myogenesis. We evaluated the electroporation efficiency and viability of C2C12 myotubes at increasing voltages (200–1300 V/cm) using propidium iodide (PI). Permeabilization of PI into the cells was voltage-dependent accounting up to 97 % efficiency at the highest voltage. High cell viability and myotube integrity were maintained until 4 days after electroporation. ECT with the cytostatic drugs bleomycin and cisplatin decreased the viability of C2C12 myoblasts and myotubes in a dose-dependent manner. However, myoblasts were more sensitive to ECT than myotubes. Increased secretion of IL-6, observed 3 days after ECT, confirming its effects on early myogenesis. Only minor effects of ECT were observed in treated myotubes. These results contribute to the safety profile of ECT in tumor treatment.

The Effect of Electrochemotherapy on Breast Cancer Cell Lines

Abstract Despite advances in treatment, breast cancer remains one of the leading causes of death, and obviously new approaches to the treatment are needed. Due to minimal side effects, unlike more aggressive forms of therapy such as chemotherapy and radiotherapy, the application of irreversible electroporation-electrochemotherapy represents a new modality in the treatment of cancer. Electrochemotherapy uses an electric field (375 V cm -1) to allow increased absorption of chemotherapeutic drugs selectively in tumor cells. Accordingly, the total dose of these agents can be significantly reduced and numerous side effects can be avoided in this way. The Real Time Cell Analysis-RTCA-xCELLigence system was used to monitor the cytotoxic effects of the treatment. The results confirmed the justification of the use of paclitaxel in chemotherapy and showed cytotoxic effects of paclitaxel which were time and dose-dependent in both cell lines. When paclitaxel was administered in combination with an electric field, in both cell lines, the results showed a greater cytotoxic effect compared to the same treatment without electrochemotherapy. MCF-7 cells are more sensitive to electrochemotherapy treatment with paclitaxel compared to MDA-MB-231. Electrochemotherapy using paclitaxel in MCF-7 cells had a 6.4-fold higher cytotoxicity compared to the treatment only with paclitaxel. The results obtained support the current knowledge of the benefits of electrochemotherapy. It has been shown that electrochemotherapy can significantly increase the effects of paclitaxel in the tested cell lines. In this way, a very high concentration of chemotherapeutics in the targeted tissue was achieved, which represents localized chemotherapy.

Effects of Electrochemotherapy on Immunologically Important Modifications in Tumor Cells.

Electrochemotherapy (ECT) is a clinically acknowledged method that combines the use of anticancer drugs and electrical pulses. Electrochemotherapy with bleomycin (BLM) can induce immunogenic cell death (ICD) in certain settings. However, whether this is ubiquitous over different cancer types and for other clinically relevant chemotherapeutics used with electrochemotherapy is unknown. Here, we evaluated in vitro in the B16-F10, 4T1 and CT26 murine tumor cell lines, the electrochemotherapy triggered changes in the ICD-associated damage-associated molecular patterns (DAMPs): Calreticulin (CRT), ATP, High Mobility Group Box 1 (HMGB1), and four immunologically important cellular markers: MHCI, MHC II, PD-L1 and CD40. The changes in these markers were investigated in time up to 48 h after ECT. We showed that electrochemotherapy with all three tested chemotherapeutics induced ICD-associated DAMPs, but the induced DAMP signature was cell line and chemotherapeutic concentration specific. Similarly, electrochemotherapy with CDDP, OXA or BLM modified the expression of MHC I, MHC II, PD-L1 and CD40. The potential of electrochemotherapy to change their expression was also cell line and chemotherapeutic concentration specific. Our results thus put the electrochemotherapy with clinically relevant chemotherapeutics CDDP, OXA and BLM on the map of ICD inducing therapies.

Repetitive Bleomycin-Based Electrochemotherapy Improves Antitumor Effectiveness in 3D Tumor Models of Conjunctival Melanoma.

Conjunctival melanoma (CM) is associated with a high rate of local recurrence and poor survival rate. Novel therapeutic options are needed to reduce recurrence rate. The objective of the study was to demonstrate the improved effectiveness of electrochemotherapy (ECT) on CM using repetitive application. Tumor spheroids of three CM cell lines (CRMM1, CRMM2, CM2005.1) were treated repetitively with ECT using the chemotherapeutic agent bleomycin on days 3, 5, and 7 of culture. Application of bleomycin alone and electroporation alone served as controls. The cytotoxic effect was analyzed on day 10 compared to untreated control using an independent t-test. The spheroid outgrowth rate was measured. CM tumor spheroid size (median value: 78%, SD: 32%) and viability (median value: 11%, SD: 11%) were dramatically reduced after repetitive ECT treatment (p-value < 0.001). Decreased proliferation capacity (down to 8%) and an increase of apoptotic cells were observed. In most repetitive ECT-treated spheroids, no viable or proliferating cells were detected. Only 33-40% of repetitive ECT-treated spheroids exhibited single outgrowing cells with a delay of time up to 38 days. Repetitive ECT application effectively induces cytotoxic effects in CM spheroids by inducing apoptosis, inhibiting proliferation and decreasing the percentage of surviving tumor cells. Thus, repetitive ECT results in improved antitumor effectiveness in CM and could be an alternative therapy option.

Defining optimal parameters to maximize the effect of electrochemotherapy on lung cancer cells whilst preserving the integrity of immune cells

Electrochemotherapy (ECT) is becoming an established therapy for melanoma and is under investigation for application in additional cancer types. One potential cancer type that may benefit from ECT is lung cancer as lung cancer treatments remain unable to deliver long-lasting treatment responses. Given the importance of the immune system in lung cancer, here we have also examined the impact of ECT on immune populations.The impact of electroporation and ECT on three human lung cancer cell lines (A549, H460, SK-MES 1), one murine cell line (LLC) and murine T cells, dendritic cells and macrophages was examined. The viability, metabolic activity and recovery potential post-treatment of all cell types was determined to evaluate the potential utility of ECT as a lung cancer treatment. Our findings demonstrate that cisplatin at 11 µM would be the suggested drug of choice when using ECT for lung cancer treatment. Our study also shows that T cells are not impacted by any tested condition, whilst dendritic cells and macrophages are significantly negatively impacted by electric field strengths surpassing 800 V/cm in vitro. Therefore, current ECT protocols (using 1000 V/cm in vivo) might need to adapted to improve viability of the immune population, thus improving therapy outcomes.

Sunitinib potentiates the cytotoxic effect of electrochemotherapy in pancreatic carcinoma cells.

BackgroundOne of the new treatment options for unresectable locally advanced pancreatic cancer is electrochemotherapy (ECT), a local ablative therapy that potentiates the entry of chemotherapeutic drugs into the cells, by the application of an electric field to the tumor. Its feasibility and safety were demonstrated in preclinical and clinical studies; however, there is a lack of preclinical studies assessing the actions of different drugs used in ECT, their mechanisms and interactions with other target drugs that are used in clinical practice.Materials and methodsThe aim of the study was to determine the cytotoxicity of two chemotherapeutic drugs usually used in ECT (bleomycin and cisplatin) in the BxPC-3 human pancreatic carcinoma cell line and evaluate the interactions of ECT with the targeted drug sunitinib. First, the cytotoxicity of ECT using both chemotherapeutics was determined. In the next part, the interactions of ECT and sunitinib were evaluated through determination of combined cytotoxicity, sunitinib targets and kinetics of cell death.ResultsThe results demonstrate that ECT is effective in pancreatic cancer cell line, especially when bleomycin is used, with the onset of cell death in the first hours after the treatment, reaching a plateau at 20 hours after the treatment. Furthermore, we provide the rationale for combining ECT with bleomycin and the targeted drug sunitinib to potentiate cytotoxicity. The combined treatment of sunitinib and ECT was synergistic for bleomycin only at the highest used concentration of bleomycin 0.14 μM, whereas with lower doses of bleomycin, this effect was not observed. The interaction of ECT and treatment with sunitinib was confirmed by course of the cell death, also indicating on synergism.ConclusionsECT and sunitinib combined treatment has clinical potential, and further studies are warranted.

(IMCS First Place Best Paper Award) A Novel Lab-on-a-Chip Microdevice for Study the Effectiveness of Electrochemotherapy

IntroductionLab-on-a-Chip systems are innovative tools which can be used in the field of life sciences. They find applications e.g. in single cell analysis or cytotoxicity tests [1]. Despite the growing popularity of the use of microsystems in biological research, there is a lack of microsystems that may be used in the electrochemotherapy (ECT) studies. ECT is a antitumor therapy, based on an application of electroporation (EP) during standard chemotherapy (CT). EP uses external electric field to form hydrophilic pores in the cells membrane. Electropores are additional migration pathway for molecules which enhanced their delivery into cells [2]. ECT allows to use lower concentration of drug and reduces side effects in comparison to standard chemotherapy [3]. We develop a Lab-on-a-Chip microsystem for cell electroporation that could be used to examine the effectiveness of chemotherapy as well as for evaluation the effectiveness of electrochemotherapy.Microsystem fabricationThe microsystem is made of polydimethylsiloxane (PDMS) and glass. Casting method was used to obtained the microchannels and microchambers in PDMS layer. Pairs of gold electrodes were arranged parallelly along the microchannel with microchambers at the distance of 2 mm. The microsystem allows simultaneous culturing of normal and tumor cells. There are four rows of microchambers for each cell line: I - cells not exposed to compound or electric field (control), II - electroporated cells not exposed to compound (control for EP), III – cells electroporated with compound (simulating condition of ECT), IV - cells incubated with compound (simulating condition of CT). In this way, it is possible to evaluate and compare the effectiveness of two types of therapeutic procedures. Method The microchip was sterilized using 70% ethanol and UV radiation. After that, the cell suspension was introduced using a peristaltic pump at a speed of 3.5 µl/min. After 24 h of incubation, the cells medium (control) and the solution of the test compound were respectively introduced into the microsystem, nextly the cells were electroporated. Cell observations were performed using an inverted fluorescence microscope. In addition, fluorescence intensity measurements of the introduced molecules were carried out using a multi-well plate reader. The AlamarBlue test was performed to determine cell viability. For this purpose, a 10% AlamarBlue solution was introduced into the microsystem, than fluorescence intensity was measured at λex=558 nm and λem= 585 nm. Results and Conclusions To determine the optimal electroporation parameters (pulse length, their number, voltage) preliminary experiments were led using propidium iodide (PI). Tests were carried out for two skin cell lines: normal HaCaT and tumor A375. Two sets of parameters were examined: 1 pulse 10 ms and 8 pulses 0.1 ms, each in three voltage variants: 150, 180 and 200V. Cell viability after electroporation was determined. It was found that there were no significant changes in the cell viability after electroporation with the voltage lower than 200V. The efficiency of PI delivery into cells was confirmed by microscopic observation as well as determined by fluorescence intensity measurements. Significantly lower PI level inside cells (at the level of 30%) using 8 pulses 0.1 ms for both cell lines was observed. The best efficiency of PI delivery (about 90%) was observed when 1 pulse of 180V lasting 10 ms was applied. In addition, cell morphology was observed and cells parameters such as: shape factor, sphericity, convexity and elongation were determined. It was confirmed that the electroporation of cells does not change their morphology. Based on the obtained results it was concluded that the optimal electroporation conditions for HaCaT and A375 cell lines are: 1 pulse 10 ms 180V.

High-Frequency and High-Voltage Asymmetric Bipolar Pulse Generator for Electroporation Based Technologies and Therapies

Currently, in high-frequency electroporation, much progress has been made but limited to research groups with custom-made laboratory prototype electroporators. According to the review of electroporators and economic evaluations, there is still an area of pulse parameters that needs to be investigated. The development of an asymmetric bipolar pulse generator with a maximum voltage of 4 kV and minimum duration time of a few hundred nanoseconds, would enable in vivo evaluation of biological effects of high-frequency electroporation pulses. Herein, from a series of most commonly used drivers and optical isolations in high-voltage pulse generators the one with optimal characteristics was used. In addition, the circuit topology of the developed device is described in detail. The developed device is able to generate 4 kV pulses, with theoretical 131 A maximal current and 200 ns minimal pulse duration, the maximal pulse repetition rate is 2 MHz and the burst maximal repetition rate is 1 MHz. The device was tested in vivo. The effectiveness of electrochemotherapy of high-frequency electroporation pulses is compared to “classical” electrochemotherapy pulses. In vivo electrochemotherapy with high-frequency electroporation pulses was at least as effective as with “classical” well-established electric pulses, resulting in 86% and 50% complete responses, respectively. In contrast to previous reports, however, muscle contractions were comparable between the two protocols.

PARP inhibitor olaparib has a potential to increase the effectiveness of electrochemotherapy in BRCA1 mutated breast cancer in mice

Electrochemotherapy (ECT), a local therapy, has different effectiveness among tumor types. In breast cancer, its effectiveness is low; therefore, combined therapies are needed. The aim of our study was to combine ECT with PARP inhibitor olaparib, which could inhibit the repair of bleomycin or cisplatin induced DNA damage and potentiate the effectiveness of ECT. The effects of combined therapy were studied in BRCA1 mutated (HCC1937) and non-mutated (HCC1143) triple negative breast cancer cell lines. Therapeutic effectiveness was studied in 2D and 3D cell cultures and in vivo on subcutaneous HCC1937 tumor model in mice. The underlying mechanism of combined therapy was determined with the evaluation of γH2AX foci. Combined therapy of ECT with bleomycin and olaparib potentiated the effectiveness of ECT in BRCA1 mutated HCC1937, but not in non-mutated HCC1143 cells. The combined therapy had a synergistic effect, which was due to the increased number of DNA double strand breaks. Addition of olaparib to ECT with bleomycin in vivo in HCC1937 tumor model had only minimal effect, indicating repetitive olaparib treatment would be needed. This study demonstrates that DNA repair inhibiting drugs, like olaparib, have the potential to increase the effectiveness of ECT with bleomycin.

Potentiation of electrochemotherapy effectiveness by immunostimulation with IL-12 gene electrotransfer in mice is dependent on tumor immune status

Electrochemotherapy (ECT) exhibits high therapeutic effectiveness in the clinic, achieving up to 80% local tumor control but without a systemic (abscopal) effect. Therefore, we designed a combination therapy consisting of ECT via intratumoral application of bleomycin, oxaliplatin or cisplatin with peritumoral gene electrotransfer of a plasmid encoding interleukin-12 (p. t. IL-12 GET). Our hypothesis was that p. t. IL-12 GET potentiates the effect of ECT on local and systemic levels and that the potentiation varies depending on tumor immune status. Therefore, the combination therapy was tested in three immunologically different murine tumor models. In poorly immunogenic B16F10 melanoma, IL-12 potentiated the antitumor effect of ECT with biologically equivalent low doses of cisplatin, oxaliplatin or bleomycin. The most pronounced potentiation was observed after ECT using cisplatin, resulting in a complete response rate of 38% and an abscopal effect. Compared to B16F10 melanoma, better responsiveness to ECT was observed in more immunogenic 4 T1 mammary carcinoma and CT26 colorectal carcinoma. In both models, p. t. IL-12 GET did not significantly improve the therapeutic outcome of ECT using any of the chemotherapeutic drugs. Collectively, the effectiveness of the combination therapy depends on tumor immune status. ECT was more effective in more immunogenic tumors, but GET exhibited greater contribution in less immunogenic tumors. Thus, the selection of the therapy, namely, either ECT alone or combination therapy with p. t. IL-12, should be predominantly based on tumor immune status.

Investigation of safety for electrochemotherapy and irreversible electroporation ablation therapies in patients with cardiac pacemakers

BackgroundThe effectiveness of electrochemotherapy of tumors (ECT) and of irreversible electroporation ablation (IRE) depends on different mechanisms and delivery protocols. Both therapies exploit the phenomenon of electroporation of the cell membrane achieved by the exposure of the cells to a series of high-voltage electric pulses. Electroporation can be fine-tuned to be either reversible or irreversible, causing the cells to either survive the exposure (in ECT) or not (in IRE), respectively. For treatment of tissues located close to the heart (e.g., in the liver), the safety of electroporation-based therapies is ensured by synchronizing the electric pulses with the electrocardiogram. However, the use of ECT and IRE remains contraindicated for patients with implanted cardiac pacemakers if the treated tissues are located close to the heart or the pacemaker. In this study, two questions are addressed: can the electroporation pulses interfere with the pacemaker; and, can the metallic housing of the pacemaker modify the distribution of electric field in the tissue sufficiently to affect the effectiveness and safety of the therapy?ResultsThe electroporation pulses induced significant changes in the pacemaker ventricular pacing pulse only for the electroporation pulses delivered during the pacing pulse itself. No residual effects were observed on the pacing pulses following the electroporation pulses for all tested experimental conditions. The results of numerical modeling indicate that the presence of metal-encased pacemaker in immediate vicinity of the treatment zone should not impair the intended effectiveness of ECT or IRE even when the casing is in direct contact with one of the active electrodes. Nevertheless, the contact between the casing and the active electrode should be avoided due to significant tissue heating at the site of the other active electrode for the IRE protocol and may cause the pulse generator to fail to deliver the pulses due to excessive current draw.ConclusionsThe observed effects of electroporation pulses delivered in close vicinity of the pacemaker or its electrodes do not indicate adverse consequences for either the function of the pacemaker or the treatment outcome. These findings should contribute to making electroporation-based treatments accessible also to patients with implanted cardiac pacemakers.

Cobalt and ruthenium complexes with pyrimidine based schiff base: Synthesis, characterization, anticancer activities and electrochemotherapy efficiency

In this study, a new Schiff base ligand and its two M(II) complexes [CoCl·L(H2O)2]·2H2O, [RuCl(p-cymene)L] were synthesized. The structural features were confirmed from their micro analytical, IR, UV–Vis., 1H–13C NMR, TGA, X-ray diffraction analysis, mass spectral data and magnetic susceptibility measurements. The Co(II) and Ru(II) complexes displayed an octahedral geometry. In vitro anticancer activities of the Schiff base, Co(II) and Ru(II) complexes were evaluated on the human colon cancer cell line (Caco-2) and biocompatibility characteristics were determined in the l-929 (normal fibroblast cells) cell line by using the MTT assay. Furthermore, we examined the effectiveness of electrochemotherapy (ECT) on cytotoxic activities of these compounds in Caco-2 cancer cell line. According to the findings of the study, Co(II) and Ru(II) complexes showed considerable anticancer properties in the Caco-2 colon cancer cells; however, the ligand did not show significant anticancer activity. It was determined that the combined application of electroporation (EP)+complexes were much more effective than the application of complexes alone in the treatment of Caco-2 colon cancer cells. In a conclusion, the Co(II) and Ru(II) complexes, which showed significant anticancer activity in Caco-2 colon cancer cells, increased cytotoxicity levels by 2.07 and 2.12, respectively in their combined applications with EP. These complexes can be developed as chemotherapeutic agents for colon cancer treatment and can yield promising results when used in ECT.

To breathe or not to breathe? Hypoxia after pulsed-electric field treatment reduces the effectiveness of electrochemotherapy in vitro

Bleomycin, which is the most widely used drugs in electrochemotherapy, requires oxygen to be able to make single- or double-strand brakes in DNA. However, the concentration of oxygen in tumours can be lower than 1%. The aim of this study was to find out whether oxygen concentration in the medium in which cells loaded with bleomycin are incubated, affects the effectiveness of electrochemotherapy in vitro. Experiments were carried out on mouse hepatoma MH-22A cells. Cells were loaded with bleomycin by using a single square-wave electric pulse (2 kV/cm, 100 μs) under normoxic conditions, seeded into Petri dishes, and grown under normoxic and hypoxic conditions. Cell viability was determined by means of a colony-forming assay. We demonstrated that when cells loaded with bleomycin were incubated in hypoxia (0.2% O2), up to 5.3-fold higher concentrations of bleomycin were needed to kill them in comparison with cells grown in normoxia (18.7% O2).

A Novel Lab-on-a-Chip Microdevice for Study the Effectiveness of Electrochemotherapy

Introduction Lab-on-a-Chip systems are innovative tools which can be used in the field of life sciences. They find applications e.g. in single cell analysis or cytotoxicity tests [1]. Despite the growing popularity of the use of microsystems in biological research, there is a lack of microsystems that may be used in the electrochemotherapy (ECT) studies. ECT is a antitumor therapy, based on an application of electroporation (EP) during standard chemotherapy (CT). EP uses external electric field to form hydrophilic pores in the cells membrane. Electropores are additional migration pathway for molecules which enhanced their delivery into cells [2]. ECT allows to use lower concentration of drug and reduces side effects in comparison to standard chemotherapy [3]. We develop a Lab-on-a-Chip microsystem for cell electroporation that could be used to examine the effectiveness of chemotherapy as well as for evaluation the effectiveness of electrochemotherapy. Microsystem fabrication The microsystem is made of polydimethylsiloxane (PDMS) and glass. Casting method was used to obtained the microchannels and microchambers in PDMS layer. Pairs of gold electrodes were arranged parallelly along the microchannel with microchambers at the distance of 2 mm. The microsystem allows simultaneous culturing of normal and tumor cells. There are four rows of microchambers for each cell line: I - cells not exposed to compound or electric field (control), II - electroporated cells not exposed to compound (control for EP), III – cells electroporated with compound (simulating condition of ECT), IV - cells incubated with compound (simulating condition of CT). In this way, it is possible to evaluate and compare the effectiveness of two types of therapeutic procedures. Method The microchip was sterilized using 70% ethanol and UV radiation. After that, the cell suspension was introduced using a peristaltic pump at a speed of 3.5 µl/min. After 24 h of incubation, the cells medium (control) and the solution of the test compound were respectively introduced into the microsystem, nextly the cells were electroporated. Cell observations were performed using an inverted fluorescence microscope. In addition, fluorescence intensity measurements of the introduced molecules were carried out using a multi-well plate reader. The AlamarBlue test was performed to determine cell viability. For this purpose, a 10% AlamarBlue solution was introduced into the microsystem, than fluorescence intensity was measured at λex=558 nm and λem= 585 nm. Results and Conclusions To determine the optimal electroporation parameters (pulse length, their number, voltage) preliminary experiments were led using propidium iodide (PI). Tests were carried out for two skin cell lines: normal HaCaT and tumor A375. Two sets of parameters were examined: 1 pulse 10 ms and 8 pulses 0.1 ms, each in three voltage variants: 150, 180 and 200V. Cell viability after electroporation was determined. It was found that there were no significant changes in the cell viability after electroporation with the voltage lower than 200V. The efficiency of PI delivery into cells was confirmed by microscopic observation as well as determined by fluorescence intensity measurements. Significantly lower PI level inside cells (at the level of 30%) using 8 pulses 0.1 ms for both cell lines was observed. The best efficiency of PI delivery (about 90%) was observed when 1 pulse of 180V lasting 10 ms was applied. In addition, cell morphology was observed and cells parameters such as: shape factor, sphericity, convexity and elongation were determined. It was confirmed that the electroporation of cells does not change their morphology. Based on the obtained results it was concluded that the optimal electroporation conditions for HaCaT and A375 cell lines are: 1 pulse 10 ms 180V.

Cell Electropermeabilisation Enhancement by Non-Thermal-Plasma-Treated PBS.

The effectiveness of electrochemotherapy (ECT) in local eradication of tumours in human and veterinary medicine has been proven. ECT consists of increasing the uptake of cytotoxic drugs by means of pulsed electric fields (PEFs) that transiently permeabilise the cell membrane. Still, this tumour treatment includes some drawbacks that are linked to the characteristics of the intense electric pulses (EPs) used. Meanwhile, the emerging field of cancer therapies that are based on the application of non-thermal plasmas (NTP) has recently garnered interest because of their potentialities as rich sources of reactive species. In this work, we investigated the potential capabilities of the combined application of indirect NTP treatment and microsecond PEFs (µsPEFs) to outperform in vitro cell electropermeabilisation, the basis of ECT. Thus, phosphate-buffered saline (PBS) was plasma-treated (pPBS) and used afterwards to explore the effects of its combination with µsPEFs. Analysis of two different cell lines (DC-3F Chinese hamster lung fibroblasts and malignant B16-F10 murine melanoma cells), by flow cytometry, revealed that this combination resulted in significant increases of the level of cell membrane electropermeabilisation, even at very low electric field amplitude. The B16-F10 cells were more sensitive to the combined treatment than DC-3F cells. Importantly, the percentage of permeabilised cells reached values similar to those of cells exposed to classical electroporation field amplitude (1100 V/cm) when the cells were treated with pPBS before and after being exposed only to very low PEF amplitude (600 V/cm). Although the level of permeabilisation of the cells that are treated by the pPBS and the PEFs at 600 V/cm is lower than the level reached after the exposure to µsPEFs alone at 1100 V/cm, the combined treatment opens the possibility to reduce the amplitude of the EPs used in ECT, potentially allowing for a novel ECT with reduced side-effects.

Use of electrochemotherapy in women with vulvar cancer to improve quality-of-life in the palliative setting: a meta-analysis

IntroductionElectrochemotherapy involves the use of transient tumor permeabilization via electric pulses in combination with low-dose chemotherapeutic agents. It has recently emerged as an alternative treatment modality in vulvar cancer. The...

Quantitative proteomic analysis of enhanced cellular effects of electrochemotherapy with Cisplatin in triple-negative breast cancer cells

Due to the lack of the three main receptors, triple negative breast cancer (TNBC) is refractive to standard chemotherapy. Hence, alternate therapies are needed. TNBCs utilize glycolysis, which heightens their growth, proliferation, invasiveness, chemotherapeutic resistance and poor therapeutic response. This calls for novel therapeutic strategies to target these metabolic vulnerabilities present in TNBC. Electroporation-mediated chemotherapy, known as electrochemotherapy (ECT) is gaining momentum as an attractive alternative. However, its molecular mechanisms need better understanding. Towards this, label-free quantitative proteomics is utilized to gain insight into the anticancer mechanisms of ECT using electrical pulses (EP) and Cisplatin (CsP) on MDA-MB-231, human TNBC cells. The results indicate that EP + CsP significantly downregulated 14 key glycolysis proteins (including ENO1, LDHA, LDHB, ACSS2, ALDOA, and PGK1), compared to CsP alone. EP + CsP caused a switch in the metabolism with upregulation of 34 oxidative phosphorylation pathway proteins and 18 tricarboxylic acid (TCA) cycle proteins compared to CsP alone, accompanied by the upregulation of proteins linked to several metabolic reactions, which produce TCA cycle intermediates. Moreover, EP + CsP promoted multiple pathways to cause 1.3-fold increase in the reactive oxygen species concentration and induced apoptosis. The proteomics results correlate well with cell viability, western blot, and qPCR data. While some effects were similar for EP, more comprehensive and long-lasting effects were observed for EP + CsP, which demonstrate the potential of EP + CsP against TNBC cells.

Effect of electrochemotherapy with betulinic acid or cisplatin on regulation of heat shock proteins in metastatic human carcinoma cells invitro.

Betulinic acid (BTA) is naturally occurring triterpene that has received interest as a novel therapeutic substance with cytotoxicity towards a number of cancer cell lines. Despite the wide spectrum of biological and pharmacological effects, its effect may be limited its lipophobic properties. Therefore, strategies to improve the access of BTA to the cells are required to enhance the anticancer effects. Electroporation (EP) enables increased inflow of drugs into cancer cells, even at low doses, which may reduce the side effects caused by high doses of chemotherapy. The potential application of BTA in electrochemotherapy (ECT) in metastatic type of cancers was investigated in the present study. The efficacy of BTA with EP was estimated using a cell survival assay (MTT assay), microscopical morphology analysis and the immunocytochemical expression of heat shock proteins (HSPs). HSPs are molecules that protect the cell from harmful environmental, chemical and physical stresses, and ensure cell survival, recovery and proper functioning. HSP expression is induced various stress factors. Therefore, the expression of HSP27 and HSP70 was evaluated after cells were exposed to an external pulsed electric field and anticancer drugs. Facilitated drug delivery and the anticancer effect on metastatic tumor cells were evaluated in vitro. The effect of BTA was compared with cisplatin (CP), a standard cytostatic agent. Two different metastatic cancer cell lines were used, an ovary adenocarcinoma cell line (SW626) and melanoma cell line (Me45). BTA combined with EP exhibited similar efficacy to CP with EP after 24 and 48 h in SW626 and Me45 cancer cells. Me45 cells also had high HSP27 and low HSP70 immunosignals post‑ECT treatment. ECT caused increased expression of HSP27 and HSP70 proteins in SW626 cells, which were less sensitive to ECT than the Me45 melanoma cell line. The results indicate that BTA may be efficiently applied instead of CP in ECT approaches, but its activity differs between tumor cell lines.

Electrochemotherapy in the treatment of locally advanced or recurrent eyelid-periocular basal cell carcinomas

There is increasing evidence about the effectiveness of electrochemotherapy (ECT) in the treatment of basal cell carcinomas in the head and neck region, although its role in the management of eyelid-periocular skin tumors has to be clarified. The aim of the present study is to evaluate the results of ECT in the treatment of locally advanced primary and recurrent eyelid-periocular skin basal cell carcinomas. Twelve patients with basal cell carcinoma involving the eyelid-periocular skin region were treated with ECT. Three patients had locally advanced primary tumors, while 9 patients had recurrent tumors. All treatments were performed according to the ESOPE guidelines, using Cliniporator TM device. All patients received bleomycin based ECT. The route of administration was intratumoral in 3 patients and intravenous in 9 patients. Tumor response was evaluated using the RECIST 1.1. criteria. ECT resulted in complete response of the periocular skin tumors in all patients. Lower eyelid ectropion was developed in 3 patients which had to be corrected surgically. ECT can be used effectively in the treatment of locally advanced or recurrent basal cell carcinomas in the eyelid-periocular skin region. Excellent tumor control can be achieved with good functional and cosmetic results without systemic adverse events with short interval follow-up.