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1.
Sci Rep ; 14(1): 12546, 2024 05 31.
Article in English | MEDLINE | ID: mdl-38822068

ABSTRACT

Nanosecond pulsed electric field (nsPEF) has emerged as a promising approach for inducing cell death in melanoma, either as a standalone treatment or in combination with chemotherapeutics. However, to date, there has been a shortage of studies exploring the impact of nsPEF on the expression of cancer-specific molecules. In this investigation, we sought to assess the effects of nsPEF on melanoma-specific MAGE (Melanoma Antigen Gene Protein Family) expression. To achieve this, melanoma cells were exposed to nsPEF with parameters set at 8 kV/cm, 200 ns duration, 100 pulses, and a frequency of 10 kHz. We also aimed to comprehensively describe the consequences of this electric field on melanoma cells' invasion and proliferation potential. Our findings reveal that following exposure to nsPEF, melanoma cells release microvesicles containing MAGE antigens, leading to a simultaneous increase in the expression and mRNA content of membrane-associated antigens such as MAGE-A1. Notably, we observed an unexpected increase in the expression of PD-1 as well. While we did not observe significant differences in the cells' proliferation or invasion potential, a remarkable alteration in the cells' metabolomic and lipidomic profiles towards a less aggressive phenotype was evident. Furthermore, we validated these results using ex vivo tissue cultures and 3D melanoma culture models. Our study demonstrates that nsPEF can elevate the expression of membrane-associated proteins, including melanoma-specific antigens. The mechanism underlying the overexpression of MAGE antigens involves the initial release of microvesicles containing MAGE antigens, followed by a gradual increase in mRNA levels, ultimately resulting in elevated expression of MAGE antigens post-experiment. These findings shed light on a novel method for modulating cancer cells to overexpress cancer-specific molecules, thereby potentially enhancing their sensitivity to targeted anticancer therapy.


Subject(s)
Exocytosis , Melanoma-Specific Antigens , Melanoma , Humans , Melanoma/metabolism , Melanoma/pathology , Melanoma/genetics , Melanoma/immunology , Cell Line, Tumor , Melanoma-Specific Antigens/metabolism , Melanoma-Specific Antigens/genetics , Cell Proliferation , Gene Expression Regulation, Neoplastic , Antigens, Neoplasm/metabolism , Antigens, Neoplasm/genetics
2.
Pharmaceuticals (Basel) ; 16(6)2023 May 30.
Article in English | MEDLINE | ID: mdl-37375757

ABSTRACT

Breast cancer ranks among the top three most common malignant neoplasms in Poland. The use of calcium ion-assisted electroporation is an alternative approach to the classic treatment of this disease. The studies conducted in recent years confirm the effectiveness of electroporation with calcium ions. Electroporation is a method that uses short electrical pulses to create transitional pores in the cell membrane to allow the penetration of certain drugs. The aim of this study was to investigate the antitumor effects of electroporation alone and calcium ion-assisted electroporation on human mammary adenocarcinoma cells that are sensitive (MCF-7/WT) and resistant to doxorubicin (MCF-7/DOX). The cell viability was assessed using independent tests: MTT and SRB. The type of cell death after the applied therapy was determined by TUNEL and flow cytometry (FACS) methods. The expression of Cav3.1 and Cav3.2 proteins of T-type voltage-gated calcium channels was assessed by immunocytochemistry, and changes in the morphology of CaEP-treated cells were visualized using a holotomographic microscope. The obtained results confirmed the effectiveness of the investigated therapeutic method. The results of the work constitute a good basis for planning research at the in vivo level and in the future to develop a more effective and safer method of breast cancer treatment for patients.

3.
Sci Rep ; 13(1): 351, 2023 01 07.
Article in English | MEDLINE | ID: mdl-36611083

ABSTRACT

Nanosecond pulsed electric fields (nsPEF) have been shown to exert anticancer effects; however, little is known about the mechanisms triggered in cancer cells by nanosecond-length pulses, especially when low, sub-permeabilization voltage is used. In this study, three human pancreatic cancer cell lines were treated with nsPEF and molecular changes at the cellular level were analyzed. Further, we assessed the efficacy of paclitaxel chemotherapy following nsPEF treatment and correlated that with the changes in the expression of multi-drug resistance (MDR) proteins. Finally, we examined the influence of nsPEF on the adhesive properties of cancer cells as well as the formation and growth of pancreatic cancer spheroids. Cell line response differed with the application of a 200 ns, 100 pulses, 8 kV/cm, 10 kHz PEF treatment. PEF treatment led to (1) the release of microvesicles (MV) in EPP85-181RDB cells, (2) electropermeabilization in EPP85-181RNOV cells and (3) cell shrinkage in EPP85-181P cells. The release of MV's in EPP85-181RDB cells reduced the membrane content of P-gp and LRP, leading to a transient increase in vulnerability of the cells towards paclitaxel. In all cell lines we observed an initial reduction in size of the cancer spheroids after the nsPEF treatment. Cell line EPP85-181RNOV exhibited a permanent reduction in the spheroid size after nsPEF. We propose a mechanism in which the surface tension of the membrane, regulated by the organization of actin fibers, modulates the response of cancer cells towards nsPEF. When a membrane's surface tension remains low, we observed some cells form protrusions and release MVs containing MDR proteins. In contrast, when cell surface tension remains high, the cell membrane is being electroporated. The latter effect may be responsible for the reduced tumor growth following nsPEF treatment.


Subject(s)
Drug Resistance, Multiple , Pancreatic Neoplasms , Humans , Cell Line , Cell Membrane/metabolism , Electroporation , Pancreatic Neoplasms/therapy , Pancreatic Neoplasms/metabolism , Pancreatic Neoplasms
4.
Bioelectrochemistry ; 150: 108356, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36566573

ABSTRACT

Ultrashort electric pulses in the nanosecond range (nsPEF) can affect extra- and intracellular lipid structures and can also alternate cell functioning reversibly and irreversibly. Several of the nsPEF effects are due to the abrupt rise in intracellular free calcium levels and calcium ions influx from the outside. Calcium is one of the most important factors in cell proliferation, differentiation, and cell death (apoptosis or necrosis). Manipulating calcium levels using electroporation can have different effects on normal and malignant cells. This study aimed to examine the impact of nsPEFs, combined with 1 mM Ca2+ in human colon adenocarcinoma cell lines: sensitive- LoVo and drug resistant-LoVoDX. In this study 200 pulses of 10 ns and high voltage (12.5-50 kVcm-1) were used. Cell viability was determined by MTT and clonogenic assay. Proteasomal activity, GSH/GSSG assay, ROS production, and PALS-1 protein were evaluated as oxidative stress markers and protein damage. Cell morphology was visualized by AFM, SEM, and confocal microscopy imaging. The results revealed that nsPEF with 1 mM Ca2+ is cytotoxic, particularly for LoVoDX cells, and safe for normal cells. NsPEF provoked ROS release, altered cell polarity, and destabilized cell morphology. These results can be important for future protocols for colon adenocarcinoma using calcium nsPEF.


Subject(s)
Adenocarcinoma , Colonic Neoplasms , Humans , Reactive Oxygen Species/metabolism , Calcium/metabolism , Colonic Neoplasms/metabolism , Cell Membrane/metabolism , Electroporation/methods , Drug Resistance
5.
Adv Clin Exp Med ; 31(9): 1049-1053, 2022 09.
Article in English | MEDLINE | ID: mdl-36135813

ABSTRACT

BACKGROUND: Annually, approx. 4000 patients are diagnosed with pancreatic cancer in Poland, and the number of deaths is close to the number of diagnoses. Such a high morbidity/mortality ratio is caused by a high percentage of unresectable lesions (about 80%) and chemoresistance, which, among other things, is due to the specific desmoplastic environment. Currently, there are 2 main systemic treatment regimens for pancreatic cancer: FOLFIRINOX (which is a combination of folic acid, fluorouracil (5-FU), irinotecan, and oxaliplatin) and combined treatment with nab-paclitaxel plus gemcitabine (NPXL+GMC). OBJECTIVES: In order to increase the effectiveness of systemic treatments for individual patients, cell lines derived from resected pancreatic tumors were developed and their chemosensitivity to various agents was examined. The hypothesis was that patients may benefit from individualization of chemotherapy. MATERIAL AND METHODS: Patients with histopathologically confirmed pancreatic cancer were operated on using irreversible electroporation (IRE) procedure. After isolating and establishing individual cell lines, chemosensitivity to 5-FU, GMC and NPXL was determined using MTT assay in primary and metastatic cell cultures. RESULTS: Three primary cell lines were isolated for the prediction of chemosensitivity. Gemcitabine was shown to be more effective at lower doses compared to 5-FU, while NPXL was more effective than 5-FU, and both of these were less effective in metastatic cells. Pancreatic cancer cell chemoresistance was confirmed in stage IV. CONCLUSION: Determination of chemosensitivity profiles using cell lines may help in the selection of systemic treatments for individual patients. This method can be the basis for a personalized planned chemotherapeutic protocol.


Subject(s)
Pancreatic Neoplasms , Antineoplastic Combined Chemotherapy Protocols/pharmacology , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Fluorouracil/pharmacology , Fluorouracil/therapeutic use , Humans , Irinotecan/pharmacology , Irinotecan/therapeutic use , Leucovorin/therapeutic use , Oxaliplatin/pharmacology , Oxaliplatin/therapeutic use , Pancreatic Neoplasms/drug therapy , Pancreatic Neoplasms
6.
Front Immunol ; 13: 889950, 2022.
Article in English | MEDLINE | ID: mdl-35874714

ABSTRACT

Cancer cells possess specific properties, such as multidrug resistance or unlimited proliferation potential, due to the presence of specific proteins on their cell membranes. The release of proliferation-related proteins from the membrane can evoke a loss of adaptive ability in cancer cells and thus enhance the effects of anticancer therapy. The upregulation of cancer-specific membrane antigens results in a better outcome of immunotherapy. Moreover, cytotoxic T-cells may also become more effective when stimulated ex-vivo toward the anticancer response. Therefore, the modulation of membrane proteins may serve as an interesting attempt in anticancer therapy. The presence of membrane antigens relies on various physical factors such as temperature, exposure to radiation, or drugs. Therefore, changing the tumor microenvironment conditions may lead to cancer cells becoming sensitized to subsequent therapy. This paper focuses on the therapeutic approaches modulating membrane antigens and enzymes in anticancer therapy. It aims to analyze the possible methods for modulating the antigens, such as pharmacological treatment, electric field treatment, photodynamic reaction, treatment with magnetic field or X-ray radiation. Besides, an overview of the effects of chemotherapy and immunotherapy on the immunophenotype of cancer cells is presented. Finally, the authors review the clinical trials that involved the modulation of cell immunophenotype in anticancer therapy.


Subject(s)
Neoplasms , Humans , Immunotherapy/methods , Neoplasms/pathology , T-Lymphocytes, Cytotoxic , Tumor Microenvironment
7.
Bioelectrochemistry ; 147: 108199, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35841647

ABSTRACT

Catechin is a bioflavonoid known for its anti-cancer properties. In the present study, we combined theoretical and experimental approaches to reveal the potential of catechin application in the electroporation (EP) or electrochemotherapy (ECT) of pancreatic cancer cells. The molecular dynamics simulations were implemented to examine the interactions of catechin with a model of a membrane, its influence on the membrane's thickness, and the impact of the catechin-membrane interaction on the pore formation. The data were confronted with experimental measurement of the threshold electric field required for permeabilization of pancreatic cancer cells to a fluorescent dye YO-PRO-1. Further, we examined the influence of catechin on cell viability following electroporation with cisplatin or calcium ions. Finally, we investigated the catechin impact on four proteins associated with multidrug resistance: P-glycoprotein, MRP1, BCRP, and LRP. We demonstrated that catechin may boost the effects of electroporation through various mechanisms: i) increasing the cell permeability prior to electroporation ii) increasing the electroporation threshold iii) sensitization of cells to chemotherapeutic compounds. We showed that catechin incubation influences mRNA levels and mitigates the immunoreactivity of Pgp, MRP1, BCRP, and LRP but these changes did not translate to the efficacy of electrochemotherapy.


Subject(s)
Catechin , Electrochemotherapy , Pancreatic Neoplasms , ATP Binding Cassette Transporter, Subfamily B/therapeutic use , ATP Binding Cassette Transporter, Subfamily G, Member 2 , Catechin/pharmacology , Catechin/therapeutic use , Cell Line, Tumor , Electroporation , Humans , Neoplasm Proteins , Pancreatic Neoplasms/drug therapy , Pancreatic Neoplasms
8.
Int J Mol Sci ; 22(20)2021 Oct 16.
Article in English | MEDLINE | ID: mdl-34681827

ABSTRACT

Two generations of positively charged poly(amidoamine) dendrimers (PAMAMs) were selected for study as potential carriers for the anticancer drug 5-fluorouracil (5FU), a drug primarily used in the treatment of colorectal cancer. Analytical techniques, such as UV-Vis spectrophotometry, NMR Spectroscopy and Laser Doppler Velocimetry (LDV), have shown that the most critical factor determining the formation of a PAMAM-5FU complex is the starting components' protonation degree. The tests confirmed the system's ability to attach about 20 5FU molecules per one dendrimer molecule for the G4PAMAM dendrimer and about 25 molecules for the G6PAMAM dendrimer, which gives a system yield of 16% for the fourth generation and 5% for sixth generation dendrimers. Additionally, using the QCM-D method, the adsorption efficiency and the number of drug molecules immobilized in the dendrimer structure were determined. A new aspect in our study was the determination of the change in zeta potential (ζ) induced by the immobilization of 5FU molecules on the dendrimer's outer shell and the importance of this effect in the direct contact of the carrier with cells. Cytotoxicity tests (resazurin reduction and MTS tests) showed no toxicity of dendrimers against mouse fibroblast cells (L929) and a significant decrease in cell viability in the case of four human malignant cell lines: malignant melanoma (A375), glioblastoma (SNB-19), prostate cancer (Du-145) and colon adenocarcinoma (HT-29) during incubation with PAMAM-5FU complexes. The purpose of our work was to investigate the correlation between the physicochemical properties of the carrier and active substance and the system efficiency and optimizing conditions for the formation of an efficient system based on PAMAM dendrimers as nanocarriers for 5-fluorouracil. An additional aspect was to identify potential application properties of the complexes, as demonstrated by cytotoxicity tests.


Subject(s)
Chemical Phenomena , Dendrimers/chemistry , Dendrimers/pharmacology , Fluorouracil/chemistry , Fluorouracil/pharmacology , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Cell Line , Drug Carriers/chemistry , Drug Carriers/pharmacology , Drug Delivery Systems/methods , Humans , Mice , Nanomedicine/methods , Nanostructures/chemistry , Nanotechnology
9.
Molecules ; 26(7)2021 Mar 25.
Article in English | MEDLINE | ID: mdl-33806009

ABSTRACT

Modifications of the composition or organization of the cancer cell membrane seem to be a promising targeted therapy. This approach can significantly enhance drug uptake or intensify the response of cancer cells to chemotherapeutics. There are several methods enabling lipid bilayer modifications, e.g., pharmacological, physical, and mechanical. It is crucial to keep in mind the significance of drug resistance phenomenon, ion channel and specific receptor impact, and lipid bilayer organization in planning the cell membrane-targeted treatment. In this review, strategies based on cell membrane modulation or reorganization are presented as an alternative tool for future therapeutic protocols.


Subject(s)
Cell Membrane , Drug Delivery Systems , Neoplasms , Cell Membrane/metabolism , Cell Membrane/pathology , Humans , Neoplasms/drug therapy , Neoplasms/metabolism , Neoplasms/pathology
10.
Bioelectrochemistry ; 140: 107806, 2021 Aug.
Article in English | MEDLINE | ID: mdl-33819839

ABSTRACT

Photodynamic therapy (PDT) and electrochemotherapy (ECT) are two methods designed to enhance the anticancer potential of various drugs. Various clinical trials proved the efficacy of both ECT and PDT in melanoma treatment. Curcumin is a natural polyphenolic compound with high anticancer potential against melanoma due to its light absorption properties and toxicity towards cancer cells; however, high reactivity and amphipathic structure of curcumin are limiting its utility. This study aimed to propose the most effective protocol for antimelanoma combination of both therapies (PDT and ECT) in the context of curcumin. The in vitro studies were carried on melanotic melanoma (A375), amelanotic melanoma (C32) and fibroblast (HGF) cell lines. In molecular dynamics studies curcumin presented the single-layer localization in the water-membrane interphase. Further, the mass spectrometry studies exposed that during the PDT treatment curcumin is degraded to vanillin, feruloylmethane, and ferulic acid. Instant ECT with curcumin followed by PDT is the most efficient approach due to its selective genotoxicity towards malignant cells. The metabolic activity of fibroblasts decreased, however, at the same time the fragmentation of DNA did not occur. Additionally, instant PDT with curcumin followed by ECT after 3 h of incubation was a therapy selective towards melanotic melanoma.


Subject(s)
Curcumin/chemistry , Curcumin/therapeutic use , Electroporation , Molecular Dynamics Simulation , Photochemotherapy/methods , Cell Line, Tumor , Combined Modality Therapy , Humans , Molecular Conformation , Water/chemistry
11.
J Pers Med ; 11(2)2021 Feb 04.
Article in English | MEDLINE | ID: mdl-33557038

ABSTRACT

PURPOSE: to assess the effect of photobiomodulation (PBM) on human gingival fibroblast proliferation. METHODS: The study was conducted using the primary cell cultures of human fibroblasts collected from systemically healthy donors. Three different laser types, Nd:YAG (1064 nm), infrared diode laser (980 nm), and prototype led laser emitting 405, 450, and 635 nm were used to irradiate the fibroblasts. Due to the patented structure of that laser, it was possible to irradiate fibroblasts with a beam combining two or three wavelengths. The energy density was 3 J/cm2, 25 J/cm2, 64 J/cm2. The viability and proliferation of cells were determined using the (Thiazolyl Blue Tetrazolium Blue) (MTT) test conducted 24, 48, and 72 h after laser irradiation. RESULTS: The highest percentage of mitochondrial activity (MA = 122.1%) was observed in the group irradiated with the 635 nm laser, with an energy density of 64 J/cm2 after 48 h. The lowest percentage of MA (94.0%) was observed in the group simultaneously irradiated with three wavelengths (405 + 450 + 635 nm). The use of the 405 nm laser at 25 J/cm2 gave similar results to the 635 nm laser. CONCLUSIONS: The application of the 635 nm and 405 nm irradiation caused a statistically significant increase in the proliferation of gingival fibroblasts.

12.
Cancers (Basel) ; 13(3)2021 Jan 22.
Article in English | MEDLINE | ID: mdl-33499085

ABSTRACT

The dynamic development of the space industry makes space flights more accessible and opens up new opportunities for biological research to better understand cell physiology under real microgravity. Whereas specialized studies in space remain out of our reach, preliminary experiments can be performed on Earth under simulated microgravity (sµg). Based on this concept, we used a 3D-clinostat (3D-C) to analyze the effect of short exposure to sµg on human keratinocytes HaCaT and melanoma cells A375 cultured on all-glass Lab-on-a-Chip (LOC). Our preliminary studies included viability evaluation, mitochondrial and caspase activity, and proliferation assay, enabling us to determine the effect of sµg on human cells. By comparing the results concerning cells cultured on LOCs and standard culture dishes, we were able to confirm the biocompatibility of all-glass LOCs and their potential application in microgravity research on selected human cell lines. Our studies revealed that HaCaT and A375 cells are susceptible to simulated microgravity; however, we observed an increased caspase activity and a decrease of proliferation in cancer cells cultured on LOCs in comparison to standard cell cultures. These results are an excellent basis to conduct further research on the possible application of LOCs systems in cancer research in space.

13.
Molecules ; 25(22)2020 Nov 19.
Article in English | MEDLINE | ID: mdl-33227916

ABSTRACT

Electroporation, applied as a non-thermal ablation method has proven to be effective for focal prostate treatment. In this study, we performed pre-clinical research, which aims at exploring the specific impact of this so-called calcium electroporation on prostate cancer. First, in an in-vitro study of DU 145 cell lines, microsecond electroporation (µsEP) parameters were optimized. We determined hence the voltage that provides both high permeability and viability of these prostate cancer cells. Subsequently, we compared the effect of µsEP on cells' viability with and without calcium administration. For high-voltage pulses, the cell death's mechanism was evaluated using flow-cytometry and confocal laser microscopy. For lower-voltage pulses, the influence of electroporation on prostate cancer cell mobility was studied using scratch assays. Additionally, we applied calcium-binding fluorescence dye (Fluo-8) to observe the calcium uptake dynamic with the fluorescence microscopy. Moreover, the molecular dynamics simulation visualized the process of calcium ions inflow during µsEP. According to our results calcium electroporation significantly decreases the cells viability by promoting apoptosis. Furthermore, our data shows that the application of pulsed electric fields disassembles the actin cytoskeleton and influences the prostate cancer cells' mobility.


Subject(s)
Adenocarcinoma/pathology , Calcium/metabolism , Electroporation/methods , Prostatic Neoplasms/pathology , Actins/metabolism , Apoptosis , Caspase 3/metabolism , Cell Death , Cell Line, Tumor , Cell Membrane Permeability , Cell Movement , Cell Survival , Extracellular Space/metabolism , Humans , Male , Molecular Dynamics Simulation , Necrosis
14.
Anticancer Res ; 40(5): 2613-2625, 2020 May.
Article in English | MEDLINE | ID: mdl-32366406

ABSTRACT

BACKGROUND/AIM: The occurrence of BRAFV600E mutation causes an up-regulation of the B-raf kinase activity leading to the stabilization of hypoxia-inducible factor 1-alpha (HIF-1α) - the promoter of the 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) enzyme. The aim of the study was to examine the effect of the (2E)-3-(3-Pyridinyl)-1-(4-pyridinyl)-2-propen-1-one (3PO), as an inhibitor of PFKFB3, on human melanoma cells (A375) with endogenous BRAFV600E mutation. MATERIALS AND METHODS: A375 cells were exposed to different concentrations of 3PO and the following tests were performed: docking, cytotoxicity assay, immunocytochemistry staining glucose uptake, clonogenic assay, holotomography imaging, and flow cytometry. RESULTS: Our studies revealed that 3PO presents a dose-dependent and time-independent cytotoxic effect and promotes apoptosis of A375 cells. Furthermore, the obtained data indicate that 3PO induces cell cycle arrest in G1/0 and glucose uptake reduction. CONCLUSION: Taking all together, our research demonstrated a here should be proapoptotic and antiproliferative effect of 3PO on A375 human melanoma cells.


Subject(s)
Enzyme Inhibitors/pharmacology , Melanoma/enzymology , Phosphofructokinase-2/antagonists & inhibitors , Pyridines/pharmacology , Apoptosis/drug effects , Caspase 3/metabolism , Caspase 8/metabolism , Catalytic Domain , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Enzyme Inhibitors/chemistry , Glucose/metabolism , Humans , Melanoma/pathology , Molecular Docking Simulation , Molecular Targeted Therapy , Phosphofructokinase-2/metabolism , Pyridines/chemistry , Tumor Stem Cell Assay
15.
Exp Ther Med ; 19(3): 2037-2044, 2020 Mar.
Article in English | MEDLINE | ID: mdl-32104263

ABSTRACT

The biological activity of chemical retraction/displacement agents in surrounding periodontal tissues is of unquestionable importance, but the activity of these agents has not been completely elucidated. In the present study, we aimed to evaluate the in vitro effects of vasoconstrictive retraction agents on primary human gingival fibroblasts (HGFs). A total of six commercial adrenergic solutions (0.05 and 0.01% HCl-epinephrine, two based on 0.05% HCl-tetrahydrozoline, 0.05% HCl-oxymetazoline, and 10% HCl-phenylephrine) and three experimental gel formulations (EG-1, EG-2, and EG-3) were used to treat primary HGFs. The biological effect of the retraction treatment on the expression of collagen types I and III was detected by performing immunocytochemical analysis. The generation of reactive oxygen species triggered by the retraction agents were evaluated by using the dichlorofluorescein (DCF) fluorescent probe. The effect of retraction agents on the expression of fibronectin was visualized by confocal laser scanning microscopy. According to the results, experimental retraction gels did not limit the expression of collagen types I and III. EG-3 even induced the synthesis of both types of collagen. The DCF assay indicated oxidative stress similar to the control cells for most of the selected retraction agents. Experimental gels did not cause degradation of the cellular shape and morphology of the primary HGFs. The proposed experimental retraction gels in the present study demonstrated higher biocompatibility with primary HGFs, suggesting their use as clinical vasoconstrictive agents for the application of gingival retraction with minimal damage to periodontal tissues.

16.
Bioelectrochemistry ; 132: 107433, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31891877

ABSTRACT

The permeabilized condition of the cell membrane after electroporation can last minutes but the underlying mechanisms remain elusive. Previous studies suggest that lipid peroxidation could be responsible for the lasting leaky state of the membrane. The present study aims to link oxidation within the plasma membrane of live cells to permeabilization by electric pulses. We have introduced a method for the detection of oxidation by ratiometric fluorescence measurements of BODIPY-C11 dye using total internal reflection fluorescence (TIRF) microscopy, limiting the signal to the cell membrane. CHO-K1 cells were cultured on glass coverslips coated with an electroconductive indium tin oxide (ITO) layer, which enabled electroporation with micro- and submicrosecond pulses. No oxidation was observed with the electric field directed towards the ITO (cathode), even at field strengths much higher than that needed for permeabilization. Oxidation was readily detectable with the opposite polarity of pulses, but with the threshold higher than the permeabilization threshold. Moreover, a decrease in the medium conductance had opposite effects on permeabilization and lipid oxidation (it enhanced the former and suppressed the latter). We conclude that lipid oxidation can indeed occur at the plasma membrane after electric pulses, but it is not the cause of lasting membrane permeabilization.


Subject(s)
Cell Membrane/metabolism , Electroporation/methods , Membrane Lipids/metabolism , Animals , Boron Compounds/metabolism , CHO Cells , Cricetulus , Oxidation-Reduction
17.
Biotechnol Appl Biochem ; 66(6): 945-954, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31476023

ABSTRACT

Primary cell cultures are challenging, but reliable model reflecting tumor response in vitro. The study was designed to examine if the increased electropermeabilization can overcame initial drug insensitivity in chondrosarcoma cells from lung metastasis. We established a primary cell culture and evaluated the cytotoxic impact of four drugs-cisplatin (CDDP), camptothecin, 2-methoxyestradiol, and leucovorin calcium (LeuCa). After determination of parameters allowing for electropermeabilization, we performed electrochemotherapy in vitro with the least toxic drugs-CDDP and LeuCa. Although combining CDDP and leucovorin together increased their toxicity and supported apoptosis, application of pulsed electric fields (PEFs) brought no advantage for their efficacy. The study emphasizes the need for introduction of primary cell cultures into studies on pulse electric fields as model frequently less sensitive to PEF-based treatments than continuous cell lines.


Subject(s)
Chondrosarcoma/pathology , Electroporation , Antineoplastic Agents/pharmacology , Cell Proliferation/drug effects , Cell Survival/drug effects , Chondrosarcoma/drug therapy , Chondrosarcoma/secondary , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Primary Cell Culture , Structure-Activity Relationship
18.
Front Pharmacol ; 10: 851, 2019.
Article in English | MEDLINE | ID: mdl-31427965

ABSTRACT

The current age of dynamic development of the space industry brings the mankind closer to routine manned space flights and space tourism. This progress leads to a demand for intensive astrobiological research aimed at improving strategies of the pharmacological protection of the human cells against extreme conditions. Although routine research in space remains out of our reach, it is worth noticing that the unique severe environment of the Earth's stratosphere has been found to mimic subcosmic conditions, giving rise to the opportunity to use the stratospheric surface as a research model for the astrobiological studies. Our study included launching into the stratosphere a balloon containing mammalian normal and cancer cells treated with various compounds to examine whether these substances are capable of protecting the cells against stress caused by rapidly varying temperature, pressure, and radiation, especially UV. Owing to oxidative stress caused by irradiation and temperature shock, we used natural compounds which display antioxidant properties, namely, catechin isolated from green tea, honokiol derived from magnolia, curcumin from turmeric, and cinnamon extract. "After-flight" laboratory tests have shown the most active antioxidants as potential agents which can minimize harmful impact of extreme conditions on human cells.

19.
Nutrients ; 11(6)2019 Jun 25.
Article in English | MEDLINE | ID: mdl-31242602

ABSTRACT

Cancers are one of the leading causes of deaths affecting millions of people around the world, therefore they are currently a major public health problem. The treatment of cancer is based on surgical resection, radiotherapy, chemotherapy or immunotherapy, much of which is often insufficient and cause serious, burdensome and undesirable side effects. For many years, assorted secondary metabolites derived from plants have been used as antitumor agents. Recently, researchers have discovered a large number of new natural substances which can effectively interfere with cancer cells' metabolism. The most famous groups of these compounds are topoisomerase and mitotic inhibitors. The aim of the latest research is to characterize natural compounds found in many common foods, especially by means of their abilities to regulate cell cycle, growth and differentiation, as well as epigenetic modulation. In this paper, we focus on a review of recent discoveries regarding nature-derived anticancer agents.


Subject(s)
Antimitotic Agents/therapeutic use , Antineoplastic Agents, Phytogenic/therapeutic use , Diet , Neoplasms/drug therapy , Topoisomerase Inhibitors/therapeutic use , Animals , Cell Cycle/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Drug Resistance, Neoplasm , Energy Metabolism/drug effects , Epigenesis, Genetic/drug effects , Humans , Neoplasms/genetics , Neoplasms/metabolism , Neoplasms/pathology
20.
Biomed Pharmacother ; 106: 1098-1107, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30119176

ABSTRACT

Photodynamic therapy (PDT) is a modern and non-invasive form of therapy, used in the treatment of non-oncological diseases as well as cancers of various types and locations. It is based on the local or systemic application of a photosensitive compound - the photosensitizer, which is accumulated in pathological tissues. The photosensitizer molecules absorb the light of the appropriate wavelength, initiating the activation processes leading to the selective destruction of the inappropriate cells. The photocytotoxic reactions occur only within the pathological tissues, in the area of photosensitizer distribution, enabling selective destruction. Over the last decade, a significant acceleration in the development of nanotechnology has been observed. The combination of photosensitizers with nanomaterials can improve the photodynamic therapy efficiency and eliminate its side effects as well. The use of nanoparticles enables achievement a targeted method which is focused on specific receptors, and, as a result, increases the selectivity of the photodynamic therapy. The object of this review is the anticancer application of PDT, its advantages and possible modifications to potentiate its effects.


Subject(s)
Drug Delivery Systems/methods , Neoplasms/drug therapy , Photochemotherapy/methods , Photosensitizing Agents/administration & dosage , Animals , Drug Carriers , Electrochemotherapy , Humans , Nanomedicine , Nanoparticles , Neoplasms/pathology , Photochemotherapy/adverse effects , Photosensitizing Agents/adverse effects
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