Piotr Suchocki

Selol-loaded magnetic nanocapsules: A new approach for hyperthermia cancer therapy

Polylactic-co-glycolic nanocapsules, loaded with nanosized magnetic particles and Selol (a selenium-based anticancer drug), were successfully prepared by the precipitation method. Maghemite (γ-Fe2O3) nanoparticles were incorporated into the nanocapsules using a highly stable ionic magnetic fluid sample. The obtained nanocapsules presented no agglomeration, negative surface charge while revealing a narrow monomodal size distribution. All the nanocapsule formulations exhibited a good physical stability at 4 °C during 3 month storage period. The in vitro antitumoral activity of Selol-magnetic nanocapsules was assessed using a murine melanoma cell line. The influence of nanocapsules on cell viability was investigated by spectrophotometric assay. The results demonstrated that Selol-loaded magnetic nanocapsules (at 100 μg/ml/5 × 109 particle/ml) showed antitumoral activity of 50% on melanoma cells (absence of magnetic field). These results clearly indicate that the loaded nanocapsules represent a novel and prom...

The Comparison of MTT and CVS Assays for the Assessment of Anticancer Agent Interactions.

Multiple in vitro tests are widely applied to assess the anticancer activity of new compounds, including their combinations and interactions with other drugs. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay is one of the most commonly used assays to assess the efficacy and interactions of anticancer agents. However, it can be significantly influenced by compounds that modify cell metabolism and reaction conditions. Therefore, several assays are sometimes used to screen for potential anticancer drugs. However, the majority of drug interactions are evaluated only with this single method. The aim of our studies was to verify whether the choice of an assay has an impact on determining the type of interaction and to identify the source of discrepancies. We compared the accuracy of MTT and CVS (crystal violet staining) assays in the interaction of two compounds characterized by similar anticancer activity: isothiocyanates (ITCs) and Selol. Confocal microscopy studies were carried out to assess the influence of these compounds on the reactive oxygen species (ROS) level, mitochondrial membrane potential, dead-to-live cell ratio and MTT-tetrazolium salt reduction rate. The MTT assay was less reliable than CVS. The MTT test of Selol and 2-oxoheptyl ITC, which affected the ROS level and MTT reduction rate, gave false negative (2-oxoheptyl ITC) or false positive (Selol) results. As a consequence, the MTT assay identified an antagonistic interaction between Selol and ITC, while the metabolism-independent CVS test identified an additive or synergistic interaction. In this paper, we show for the first time that the test assay may change the interpretation of the compound interaction. Therefore, the test method should be chosen with caution, considering the mechanism of action of the compound.

Co-nanoencapsulation of magnetic nanoparticles and selol for breast tumor treatment: in vitro evaluation of cytotoxicity and magnetohyperthermia efficacy

Antitumor activities have been described in selol, a hydrophobic mixture of molecules containing selenium in their structure, and also in maghemite magnetic nanoparticles (MNPs). Both selol and MNPs were co-encapsulated within poly(lactic-co-glycolic acid) (PLGA) nanocapsules for therapeutic purposes. The PLGA-nanocapsules loaded with MNPs and selol were labeled MSE-NC and characterized by transmission and scanning electron microscopy, electrophoretic mobility, photon correlation spectroscopy, presenting a monodisperse profile, and positive charge. The antitumor effect of MSE-NC was evaluated using normal (MCF-10A) and neoplastic (4T1 and MCF-7) breast cell lines. Nanocapsules containing only MNPs or selol were used as control. MTT assay showed that the cytotoxicity induced by MSE-NC was dose and time dependent. Normal cells were less affected than tumor cells. Cell death occurred mainly by apoptosis. Further exposure of MSE-NC treated neoplastic breast cells to an alternating magnetic field increased the antitumor effect of MSE-NC. It was concluded that selol-loaded magnetic PLGA-nanocapsules (MSE-NC) represent an effective magnetic material platform to promote magnetohyperthermia and thus a potential system for antitumor therapy.

Selenitetriglicerydes affect CYP1A1 and QR activity by involvement of reactive oxygen species and Nrf2 transcription factor

Selenitetriglycerides are a group of compounds that contain selenium (Se) (IV). In this paper, we present the results of examinations of three structurally-related selenitetriglicerydes that contain various Se concentrations: 2%, 5% and 7% Selol. The present study concentrates on the effect of Selol on phase 1 and 2 enzyme activity and the implications of free radicals and the nuclear erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway in the activity of this compound. The cytotoxic and cytostatic activities of the three kinds of Selol were evaluated; however, the cytotoxic effect was observed only for 7% Selol. Our results show that 2% Selol acts as a monofunctional inducer of phase 2 enzyme activity, and the induction is mediated by the Nrf2 transcription factor. Selol 7% acts in an opposite manner and induces phase 1 with simultaneous inhibition of phase 2 enzyme activity. The differential effect can be associated with the increase in Se content, leading to a change in the structure of the compound.

Optimization of Selenium-Enriched Mycelium of Lentinula edodes (Berk.) Pegler as a Food Supplement

Our goal was to optimize the growth conditions of submerged mycelial cultures of Lentinula edodes (Shiitake mushroom) in order to obtain a new dietary supplement enriched in selenium We designed a process technology in which mycelial cultures were cultivated in media composed of beet molasses, 10%; liquid stillage, 5%; corn steep liquor, 0.15%; and KH2PO4, 0.3%, enriched with selenium in concentrations ranging from 0 to 100 μg/mL by the addition of sodium selenite of selenic acid. Se concentrations in mycelial dry mass rose from 0.001 mg/g (mycelia cultivated in media containing no selenium) to 50 mg/g (mycelia cultivated in medium containing 100 μg Se/mL). The highest mycelial specific growth rate (0.46/day) was recorded when the concentration of selenium in the medium was lower than 20 μg/mL. In vitro, estimated Se bioavailabilities from selenized mycelium strongly depended on its preparation in a proper manner and were 60% and 82% for dried mycelium and mycelial lyophilizate, respectively. Our results suggest that these optimized culture conditions could be applied to obtain a new Se-enriched dietary supplement.

PVM/MA-shelled selol nanocapsules promote cell cycle arrest in A549 lung adenocarcinoma cells

BackgroundSelol is an oily mixture of selenitetriacylglycerides that was obtained as a semi-synthetic compound containing selenite. Selol is effective against cancerous cells and less toxic to normal cells compared with inorganic forms of selenite. However, Selol’s hydrophobicity hinders its administration in vivo. Therefore, the present study aimed to produce a formulation of Selol nanocapsules (SPN) and to test its effectiveness against pulmonary adenocarcinoma cells (A549).ResultsNanocapsules were produced through an interfacial nanoprecipitation method. The polymer shell was composed of poly(methyl vinyl ether-co-maleic anhydride) (PVM/MA) copolymer. The obtained nanocapsules were monodisperse and stable. Both free Selol (S) and SPN reduced the viability of A549 cells, whereas S induced a greater reduction in non-tumor cell viability than SPN. The suppressor effect of SPN was primarily associated to the G2/M arrest of the cell cycle, as was corroborated by the down-regulations of the CCNB1 and CDC25C genes. Apoptosis and necrosis were induced by Selol in a discrete percentage of A549 cells. SPN also increased the production of reactive oxygen species, leading to oxidative cellular damage and to the overexpression of the GPX1, CYP1A1, BAX and BCL2 genes.ConclusionsThis study presents a stable formulation of PVM/MA-shelled Selol nanocapsules and provides the first demonstration that Selol promotes G2/M arrest in cancerous cells.

In vitro cytotoxicity of Selol-loaded magnetic nanocapsules against neoplastic cell lines under AC magnetic field activation

The goals of this study are to evaluate invitro compatibility of magnetic nanomaterials and their therapeutic potential against cancer cells. Highly stable ionic magnetic fluid sample (maghemite, γ-Fe2O3) and Selol were incorporated into polymeric nanocapsules by nanoprecipitation method. The cytotoxic effect of Selol-loaded magnetic nanocapsules was assessed on murine melanoma (B16-F10) and oral squamous cell carcinoma (OSCC) cell lines following AC magnetic field application. The influence of different nanocapsules on cell viability was investigated by colorimetric MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. In the absence of AC magnetic field Selol-loaded magnetic nanocapsules, containing 100 µg/mL Selol plus 5 × 1012 particle/mL, showed antitumoral activity of about 50% on B16-F10 melanoma cells while OSCC carcinoma cells demonstrated drug resistance at all concentrations of Selol and magnetic fluid (range of 100–500 µg/mL Selol and 5 × 1012−2.5 × 1013 particle/mL). On the...

Selenitetriglycerides—Redox-active agents

BACKGROUND Human prostate cancer (hPCa) is the most commonly diagnosed cancer in elderly men and is the second leading cause of male cancer death. Data from epidemiological, eco-environmental, nutritional prevention and clinical trials suggest that selenium Se(IV) can prevent prostate cancer. Selol, a new organic semisynthetic derivative of Se(IV), is a mixture of selenitetriglycerides. This mixture is non-toxic and non-mutagenic, and after po treatment - 56-times less toxic (in mice) than sodium selenite. It exhibits strong anti-cancer activity in vitro in many cancer cell lines and can overcome the cell resistance to doxorubicin. Selol seems a promising compound for prostate cancer therapy. MATERIALS AND METHODS The aim of the present study is the evaluation of Selols influence on intracellular redox state (Eh) of prostatic tumors and the liver in androgen-dependent hPCa-bearing mice, and extracellular redox state in serum of these mice. RESULTS AND CONCLUSIONS The anticancer activity of Selol involves perturbation of the redox regulation in the androgen dependent hPCa (LNCaP) cells, but not in healthy cells. After Selol treatment, intracellular Eh has increased in tumors from -223 mV to -175 mV, while in serum it has decreased (-82 mV vs -113 mV). It shows significant changes Eh in the extra- and intracellular environment. The difference decreases from 141 mV to 62 mV. The changes suggest that a tumor cell was probably directed toward apoptosis. This is exemplified in a significant decrease in cancer tumor mass by approx. 17% after the three weeks of Selol administration.

Effects of Selol 5% supplementation on the activity or concentration of antioxidants and malondialdehyde level in the blood of healthy mice

BACKGROUND Selol is a novel organoselenium Se(IV) compound. It reveals lower potential of toxicity than sodium selenite and does not exhibit mutagenic activity. Its antioxidant and anticancer properties including overcoming cancer cell resistance to standard therapy of the drug were proven. This is the first publication describing the influence of Selol 5% on the activity of blood antioxidant status in vivo. MATERIALS AND METHODS We investigated the influence of Selol 5% short-term (24h) and long-term (28 days) administration on the activity of antioxidant enzymes, including the main selenoenzymes, in healthy mice plasma and erythrocytes. Plasma oxygen radical absorbance capacity value (ORAC) and the concentration of malonyldialdehyde (MDA) in plasma as a biomarker of oxidative stress as well as the value of selenium (Se) concentration in erythrocytes were shown. RESULTS A significant increase of the selenium dependent glutathione peroxidase (Se-GSHPx) activity in plasma and erythrocytes, plasma selenoprotein P concentration, ORAC values, and Se concentration were observed during long-term supplementation as well as after Selol 5% single-dose administration, with two distinct increases of activity a few hours after the beginning of the experiment and before its end. We found a decreased thioredoxin reductase (THRR) activity and an increased MDA level during Selol 5% long-term supplementation. Glutathione S-transferase activity (GST) remained unchanged. CONCLUSION Selol 5% supplementation in vivo affects the selenoenzymes activities as well as the antioxidant status of plasma and erythrocytes. Selol 5% is an inhibitor of thioredoxin reductase activity, which can be important in anticancer therapy.

Antimitotic effect of Selol and sodium selenate (IV) on Allium test cells

Selol is a mixture of selenitetriglycerides, obtained by esterification of pre-oxidated triglycerides from sunflower oil with selenous acid. Investigations have been carried out on the anticancer use of Selol and its antimitotic properties. The aim of our work was to compare the effect of two compounds (an organic and an inorganic one) containing selenium with oxidation number at +4: Selol and sodium selenate (IV), on plant cells of Allium test. Structural changes in cells, their mitotic activity and proportions of mitotic phases were subject to experimental analysis. Incubation in both selenium compounds led to changes in structure of chromosomes, consisting in their strong condensation and contraction (cc chromosomes), accompanied by gradual reduction or complete inhibition of cell division. High concentrations of Selol led to strong condensation of chromatin in interphase cells, while cells treated with high concentrations of sodium selenate (IV) displayed typical “pseudoapoptotic” bodies and totally fragmented nuclei. Both compounds caused inhibition of cell division; however, sodium selenate (IV) was more effective, while low concentrations of Selol stimulated division. Use of Selol resulted in changes in chromosome structure, condensation and contraction, which made it impossible to pass to further phases of cell division. The effect of sodium selenate (IV) consisted in raised prophase index, and high concentrations led to inhibition of divisions and arrest of cells at different stages of mitosis. The results obtained show markedly lower toxicity of Selol in comparison with sodium selenate, which makes it advisable to further investigate that compound as a potential safe anticancer drug.