Our Publications Database

Article 39

Noninvasive detection of changes in cells' cytosol conductivity by combining dielectrophoresis with optical tweezers

Moisescu, Mihaela Georgeta; Savopol, Tudor; Dimitriu, Liviu; Cemazar, Jaka; Kovacs, Eugenia; Radu, Mihai

Journal: Analytica Chimica ACTA

Year: 2018

Cytosol conductivity; Dielectrophoresis; Optical tweezers; Electroporation; Clausius-Mossotti factor; Murine melanoma cells

Cellular electrical properties are modulated by various physical and/or chemical stresses and detection of these changes is a challenging issue. Optical tweezers (OT) and dielectrophoresis (DEP) are frequently integrated to devices dedicated to the investigation of cells properties. Here we provide a technique to detect changes in cytosol conductivity of cells by using a combination of DEP and OT. The method was exemplified for the case of cells electroporation and is based on balancing the DEP force by a controlled OT force. We observed a decrease of the DEP force in the case of electroporated cells which was correlated to a decrease of cytosol conductivity by means of Clausius- Mossotti factor modeling. For highly stressing electroporation pulses, the cytosol conductivity drops to values close to those of the cells suspending medium. These results are consistent with those reported in the literature proving the robustness of our proposed sensing method. (C) 2018 Elsevier B.V. All rights reserved.

Article 38

Evaluation of the metastatic potential of malignant cells by image processing of digital holographic microscopy data

Violeta L Calin, Mona Mihailescu, Eugen I Scarlat, Alexandra V Baluta, Daniel Calin, Eugenia Kovacs, Tudor Savopol, Mihaela G. Moisescu

Journal: FEBS Open Bio

Year: 2017

Bimodality coefficient analysis of quantitative phase image; Clonogenic test; Digital holographic microscopy; Impedance‐based cell proliferation test; Metastatic potential; Murine B16 malignant cells

The cell refractive index has been proposed as a putative cancer biomarker of great potential, being correlated with cell content and morphology, cell division rate and membrane permeability. We used digital holographic microscopy to compare the refractive index and dry mass density of two B16 murine melanoma sublines of different metastatic potential. Using statistical methods, the distribution of phase shifts within the reconstructed quantitative phase images was analyzed by the method of bimodality coefficients. The observed correlation of refractive index, dry mass density and bimodality profile with the metastatic potential of the cells was validated by real time impedance-based assay and clonogenic tests. We suggest that the refractive index and bimodality analysis of quantitative phase image histograms could be developed as optical biomarkers useful in label-free detection and quantitative evaluation of cell metastatic potential.

Article 36

Changes in optical properties of electroporated cells as revealed by digital holographic microscopy

Violeta L Calin, Mona Mihailescu, Nicolae Mihale, Alexandra V. Baluta, Eugenia Kovacs, Tudor Savopol, Mihaela G. Moisescu

Journal: Biomedical Optics Express

Year: 2017

Digital holography; Medical optics and biotechnology; Cell analysis

Changes in optical and shape-related characteristics of B16F10 cells after electroporation were investigated using digital holographic microscopy (DHM). Bipolar rectangular pulses specific for electrochemotherapy were used. Electroporation was performed in an “off-axis” DHM set-up without using exogenous markers. Two types of cell parameters were monitored seconds and minutes after pulse train application: parameters addressing a specifically defined area of the cell (refractive index and cell height) and global cell parameters (projected area, optical phase shift profile and dry mass). The biphasic behavior of cellular parameters was explained by water and mannitol dynamics through the electropermeabilized cell membrane.

Article 35

Polygonum bistorta L. herba et flores. Polyphenols profile, antioxidant properties and cytotoxic effect on murine fibroblast cell line NIH3T3

Lucia Pîrvu, Fawzia Sha’at, Luminița Claudia Miclea, Tudor Savopol, Georgeta Neagu, Denisa Ioana Udeanu, Mihaela Georgeta Moisescu

Journal: Farmacia

Year: 2017

DOI: 10.1364/BOE.8.002222

Polygonum bistorta, Polyphenols, Antioxidant, Cytotoxicity, Fibroblast cells, NIH3T3

This work aimed to study the polyphenols content, the antioxidant activity and the potential cytotoxic effects on fibroblast
murine cells NIH3T3 of an ethanolic extract (70%, v/v) obtained from the aerial part (herba et flores) of Polygonum bistorta
L. (Polygonaceae family). The tested extract was assessed for its total phenols content: 1.15 mg gallic acid equivalents
(GAE) per 1 mL extract and showed numerous flavonoids compounds such as quercetin, kaempferol and luteolin derivates,
as well as caffeic and chlorogenic phenolic acids. The chemiluminescence (CL) tests indicated a high antioxidant capacity
(IC50 = 0.57 µg GAE/mL), superior to gallic acid (IC50 = 0.85 µg/mL) and rutin (IC50 = 2.54 µg/mL), used as reference
compounds. The cytotoxicity test demonstrated that, at concentrations over 25 µg GAE/mL extract, the ethanolic extracts
from the aerial part of Polygonum bistorta plant species have a significant toxicity on mouse embryonic fibroblast NIH3T3 cells.

Article 33

Hemorrhagic risk due to platelet dysfunction in myelodysplastic patients, correlations with anemia severity and iron overload

Popov, Viola M.; Vladareanu, Ana M.; Bumbea, Horia; Kovacs, Eugenia; Savopol, Tudor; Iordache, Maria M.; Moisescu, Mihaela G.

Journal: Blood Coagulation & Fibrinolysis

Year: 2015

DOI: 10.1097/MBC.0000000000000287 

Aggregation; Anemias; Hemorrhages; Hemosideroses; Membrane glycoproteins; Myelodysplastic syndrome; Platelet

Platelet function is influenced by changes in membrane fluidity that has an important role in the expression of platelet receptors and in modulating the activity of proteins like phospholipase C or proteinkinase C. In freshly prepared platelets, membrane fluidity modifies the aggregation/agglutination function. Reactive oxygen species (ROS) represent another important parameter involved in platelet receptor activation. There is a certain association of high levels of ROS and iron overload. Patients with hemochromatosis have low platelet aggregation induced by thrombin; little is known about the anemia and effects of iron overload on platelet activation in myelodysplastic syndromes (MDS) patients. Study of platelet membrane fluidity and ROS production changes in patients with MDS and possible correlations with altered platelet function as reflected in aggregation curves and platelet receptor expression. To find out possible correlations of fluidity of platelet membrane and ROS level with hematologic parameters and iron levels. The prospective study included 34 patients with myelodysplastic syndromes classified according to French-American-British cooperative group proposals and 29 healthy volunteers. Platelet membrane fluidity was quantified by fluorescence anisotropy measurements using the marker 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluenesulfonate. ROS production was evaluated by fluorescence measurements using 2′,7′-dichlorodihydrofluorescein diacetate. Platelet function was analyzed by optical aggregometry using the agonists adenosine diphosphate, collagen, ristocetin and epinephrine. The expression of platelet receptors CD41/ CD61, CD42a/CD42b and CD62P/CD63 was evaluated by flow cytometry. Platelet membrane fluidity in patients with MDS was similar to that of healthy volunteers and did not vary according to the risk category. Patients with MDS had increased platelet ROS production compared with the control group without statistical correlation with membrane fluidity. We found a negative correlation of ROS levels with the severity of anemia (R=-0.587, P=0.017). Platelet response was reduced in patients with MDS compared with volunteers, for all reagents. The response was different according to the risk category only in case of ristocetin or collagen. Patients with anemia presented a decreased platelet aggregation induced by collagen or ristocetin (collagen: R=0.395, P=0.003; ristocetin: R=0.420, P=0.002). The membrane fluidity of platelets from MDS patients appeared unmodified, but the ROS production was increased in all risk categories of MDS. The levels of ROS were correlated with the degree of anemia, which, in turn, had a negative impact on the platelet aggregation function induced by collagen or ristocetin. Copyright (c) 2015 Wolters Kluwer Health, Inc. All rights reserved.

Article 32

Stretching of red blood cells using an electro-optics trap

Haque, Md. Mozzammel; Moisescu, Mihaela G.; Valkai, Sandor; Der, Andras; Savopol, Tudor

Journal: Biomedical Optics Express

Year: 2015

Human Erythrocyte-Membrane; Optical Tweezers; Deformability; Manipulation; Mechanics; System

The stretching stiffness of Red Blood Cells (RBCs) was investigated using a combination of an AC dielectrophoretic apparatus and a single-beam optical tweezer. The experiments were performed at 10 MHz, a frequency high enough to avoid conductivity losses, but below the second turnover point between positive and negative dielectrophoresis. By measuring the geometrical parameters of single healthy human RBCs as a function of the applied voltage, the elastic modulus of RBCs was determined (mu = 1.80 +/- 0.5 mu N/m) and compared with similar values of the literature got by other techniques. The method is expected to be an easy-to-use, alternative tool to determine the mechano-elastic properties of living cells, and, on this basis, to distinguish healthy and diseased cells (C) 2014 Optical Society of America

Article 31

Correlation of the intracellular reactive oxygen species levels with textural properties of functionalized mesostructured silica

Laura Bajenaru, Daniela Berger, Luminita Miclea, Cristian Matei, Silviu Nastase, Cristian Andronescu, Mihaela G Moisescu, Tudor Savopol

 

Journal: Journal Of Biomedical Materials

Year: 2014

Fibroblasts; Functionalized MCM-41 silica; Intracellular reactive oxygen species; Melanocytes

Mesostructured silica is frequently used in biomedical applications, being considered nontoxic and biocompatible material, suitable for the development of drug delivery systems (DDS). Four functionalized MCM-41 silica materials with hydrophobic (methyl and vinyl) and hydrophilic (3-aminopropyl and 3-mercaptopropyl) groups were obtained by post-synthesis functionalization and characterized by small-angle X-ray diffraction, infrared spectroscopy, thermal analysis, and nitrogen adsorption-desorption isotherms. The main structural and textural parameters of the obtained silica were determined. The effect of the functionalized silica on fibroblast (NIH3T3) and melanocyte cells (B16F10) was studied with respect to the proliferation rate and the levels of reactive oxygen species (ROS). It was found that the textural properties of all samples influenced the levels of intracellular ROS and consequently, the proliferation rate. Both, healthy and malignant cells exhibited linear dependence of ROS levels with the specific surface area values, but with different response. The contribution of the methyl functionalized silica to the ROS level is apart to the general trend.

Article 30

The interaction of aminoglycosides with endothelial cells infected by Pseudomonas Aeruginosa

Savopol, Tudor ; Moraru, Roxana ; Dinn, A ; Kovacs, Eugenia ; Sajin, Gheorghe

Journal: Romanian Journal of Biophysics

Year: 2014

ISBN: 1220-515X

Aminoglycosides, Pseudomonas Aeruginosa, Endothelial Cells

The membrane organization of cultured human endothelial cells (EA.hy926) infected with Pseudomonas aeruginosa (PS) was studied in the temperature range 15–37 oC by using fluorescent depolarization and generalized polarization (GP) measurements on the cells suspensions labeled with TMA-DPH and laurdan, respectively. The opportunistic pathogen Pseudomonas aeruginosa was chosen since this pathogen is the etiologic agent of resistant and recurrent pulmonary infections in cystic fibrosis (CF) patients, who require sustained treatment with antibiotics. The effect of several aminoglycoside antibiotics – gentamicin, amikacin and kanamycin – on membrane organization of infected cultured cells was recorded. The measurements results showed that the presence of aminoglycosides cations induces a rigidity of the infected cell membrane, gentamicin being the most efficient in this respect. This effect is temperature sensitive, being much more pronounced at temperatures close to physiological range. The above information may be of use in determining the treatment regimen of a CF patient who has recurrent infections and requires antibiotics treatment for a longer time.

Article 29

Mesostructured silica matrix for irinotecan delivery systems

Nastase, Silviu; Bajenaru, Laura; Berger, Daniela; Matei, Cristian; Moisescu, Mihaela G.; Constantin, Doru; Savopol, Tudor

Journal: Central European Journal Of Chemistry

Year: 2014

DOI: 10.2478/s11532-014-0501-y 

  

 

Irinotecan; Drug delivery; Cytotoxicity; Mesoporous silica; MCM-41

Three mesostructured silica-type carriers, MCM-41 and MCM-41 functionalized by a postsynthesis grafting procedure with hydrophilic aminopropyl groups (MCM-APTES) and hydrophobic vinyl moieties (MCM-VTES), respectively, were investigated in order to elaborate drug delivery systems (DDS) for irinotecan molecules. All studied drug delivery systems exhibited higher cytotoxicity on murine embrionary fibroblastic (MEF) cells than free irinotecan at the same content of the cytostatic agent, whereas no toxicity was observed for the three unloaded carriers. The cytotoxic effect of irinotecan loaded on MCM-41-type carriers continued to increase even 24 h after ceasing the cell exposure to the drug and remained significantly higher than that of free irinotecan. The cellular uptake of silica-type hybrids was investigated by labelling MCM-APTES with Rhodamine B. In the case of the studied DDS, an endocytotic mechanism was found to be involved in the cell uptake process, and it was used to explain the cytotoxicity differences between free irinotecan and drug loaded on MCM-41-type supports.

Article 28

Setup for simultaneous microwave heating and real-time spectrofluorometric measurements in biological systems

Kohler, Sophie; Ticaud, Nicolas, Iordache, Maria-Minodora; Moisescu, Mihaela G; Savopol, Tudor; Leveque, Philippe; Arnaud-Cormos, Delia

Journal: Progress in electromagnetics research-pier

Year: 2014

Plus modern organic-synthesis, Exposure system, Laurdan fluorescence, Fields, Membranes, Model

In this paper, a delivery system allowing simultaneous microwave heating and real-time spectrofluorometric measurements in biological systems is proposed and characterized. This system is used to investigate the phase behavior of lipid bilayers from about 15 C to 45 C. The delivery system is based on an open transverse electromagnetic (TEM) cell combined with a spectrofluorometer via an optical cable system. A numerical and experimental dosimetry of the delivery system is conducted. The Specific Absorption Rate (SAR) efficiency of the system is 26.1 +/- 2.1 W/kg/W. Spectrofluorometric measurements on Laurdan labeled small unilamellar vesicles (SUVs) are carried out. Generalized polarization (GP) of the SUV’s membrane is obtained from the fluorescence intensities measured at two emission wavelengths.

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