ABSTRACT
In this work, a new approach to construct self-assembled hybrid systems based on natural PSII-enriched thylakoid membranes (PSII BBY) is demonstrated. Superfine m-WO3 NPs (≈1-2 nm) are introduced into PSII BBY. Transmission electron microscopy (TEM) measurements showed that even the highest concentrations of NPs used did not degrade the PSII BBY membranes. Using atomic force microscopy (AFM), it is shown that the organization of PSII BBY depends strongly on the concentration of NPs applied. This proved that the superfine NPs can easily penetrate the thylakoid membrane and interact with its components. These changes are also related to the modified energy transfer between the external light-harvesting antennas and the PSII reaction center, shown by absorption and fluorescence experiments. The biohybrid system shows stability at pH 6.5, the native operating environment of PSII, so a high rate of O2 evolution is expected. In addition, the light-induced water-splitting process can be further stimulated by the direct interaction of superfine WO3 NPs with the donor and acceptor sides of PSII. The water-splitting activity and stability of this colloidal system are under investigation. RESEARCH HIGHLIGHTS: The phenomenon of the self-organization of a biohybrid system composed of thylakoid membranes enriched in photosystem II and superfine WO3 nanoparticles is studied using AFM and TEM. A strong dependence of the organization of PSII complexes within PSII BBY membranes on the concentration of NPs applied is observed. This observation turns out to be crucial to understand the complexity of the mechanism of the action of WO3 NPs on modifications of energy transfer from external antenna complexes to the PSII reaction center.
Subject(s)
Nanoparticles , Thylakoids , Thylakoids/chemistry , Thylakoids/metabolism , Photosystem II Protein Complex/analysis , Photosystem II Protein Complex/metabolism , Energy Transfer , Water/analysisABSTRACT
In this work, optimized size distribution and optical properties in the colloidal synthesis of gold nanoparticles (GNPs) were obtained using a proposed ultrasonic irradiation assisted Turkevich-Frens method. The effect of three nominal ultrasound (20 kHz) irradiation powers: 60, 150, and 210 W have been analyzed as size and shape control parameters. The GNPs colloidal solutions were obtained from chloroauric acid (HAuCl4) and trisodium citrate (C6H5Na3O7·2H2O) under continuous irradiation for 1 h without any additional heat or stirring. The surface plasmon resonance (SPR) was monitored in the UV-Vis spectra every 10 min to found the optimal time for localized SPR wavelength (λLSPR), and the 210 sample procedure has reduced the λLSPR localization at 20 min, while 150 and 60 samples have showed λLSPR at 60 min. The nucleation and growth of GNPs showed changes in shape and size distribution associated with physical (cavitation, temperature) and chemical (radical generation, pH) conditions in the aqueous solution. The results showed quasi-spherical GNPs as pentakis dodecahedron (λLSPR = 560 nm), triakis icosahedron (λLSPR = 535 nm), and tetrakis hexahedron (λLSPR = 525 nm) in a size range from 12 to 16 nm. Chemical effects of ultrasound irradiation were suggested in the disproportionation process, electrons of AuCl2- are rapidly exchanged through the gold surface. After AuCl4- and Cl- were desorbed, a tetrachloroaurate complex was recycled for the two-electron reduction by citrate, aurophilic interaction between complexes AuCl2-, electrons exchange, and gold seeds, the deposition of new gold atoms on the surface promoting the growth of GNPs. These mechanisms are enhanced by the effects of ultrasound, such as cavitation and transmitted energy into the solution. These results show that the plasmonic response from the reported GNPs can be tuned using a simple methodology with minimum infrastructure requirements. Moreover, the production method could be easily scalable to meet industrial manufacturing needs.
ABSTRACT
The integration of noble metal and magnetic nanoparticles with controlled structures that can couple various specific effects to the different nanocomposite in multifunctional nanosystems have been found interesting in the field of medicine. In this work, we show synthesis route to prepare small Au nanoparticles of sizes
Subject(s)
Cinnamomum/chemistry , Ferric Compounds/chemistry , Green Chemistry Technology/methods , Hyperthermia, Induced , Magnetics , Nanoparticles/chemistry , Polyphenols/chemistry , Vanilla/chemistry , Animals , Cell Line , Mice , Nanoparticles/ultrastructure , Spectroscopy, Fourier Transform Infrared , X-Ray DiffractionABSTRACT
We present a systematic study of core-shell Au/Fe3O4 nanoparticles produced by thermal decomposition under mild conditions. The morphology and crystal structure of the nanoparticles revealed the presence of Au core of d = (6.9 ± 1.0) nm surrounded by Fe3O4 shell with a thickness of ~3.5 nm, epitaxially grown onto the Au core surface. The Au/Fe3O4 core-shell structure was demonstrated by high angle annular dark field scanning transmission electron microscopy analysis. The magnetite shell grown on top of the Au nanoparticle displayed a thermal blocking state at temperatures below TB = 59 K and a relaxed state well above TB. Remarkably, an exchange bias effect was observed when cooling down the samples below room temperature under an external magnetic field. Moreover, the exchange bias field (HEX) started to appear at T~40 K and its value increased by decreasing the temperature. This effect has been assigned to the interaction of spins located in the magnetically disordered regions (in the inner and outer surface of the Fe3O4 shell) and spins located in the ordered region of the Fe3O4 shell.
ABSTRACT
We present a proof of concept on the use of thermomagnetic polymer films (TMFs) as heating devices for magnetic hyperthermia in vitro. The TMFs were prepared through spray assisted layer-by-layer assembly of polysaccharides and magnetic iron oxide nanoparticles, yielding an alternate magnetic-polymer multilayer structure. By applying a remote alternating magnetic field (AMF) (f = 180 kHz; H = 35 kA m-1) we increased the temperature of the TMFs in a thickness-dependent way, up to 12 °C within the first 5 minutes. To test our films as heating substrates for magnetic hyperthermia, a series of in vitro experiments were designed using human neuroblastoma SH-SY5Y cells, known by their tolerance to thermal stress. The application of two AMF cycles (30 minutes each) showed that the exogenous magnetic hyperthermia resulted in an 85% reduction of cell viability. This capacity of the TMFs of hyperthermia-mediated cell killing using a remote AMF opens new options for the treatment of small and superficial tumor lesions by means of remotely-triggered magnetic hyperthermia.
ABSTRACT
In this research work, DEXTRAN- and polyethylene glycol (PEG)-coated iron-oxide superparamagnetic nanoparticles were synthetized and their cytotoxicity and biodistribution assessed. Well-crystalline hydrophobic Fe3 O4 SPIONs were formed by a thermal decomposition process with d = 18 nm and σ = 2 nm; finally, the character of SPIONs was changed to hydrophilic by a post-synthesis procedure with the functionalization of the SPIONs with PEG or DEXTRAN. The nanoparticles present high saturation magnetization and superparamagnetic behavior at room temperature, and the hydrodynamic diameters of DEXTRAN- and PEG-coated SPIONs were measured as 170 and 120 nm, respectively. PEG- and DEXTRAN-coated SPIONs have a Specific Power Absorption SPA of 320 and 400 W/g, respectively, in an ac magnetic field with amplitude of 13 kA/m and frequency of 256 kHz. In vitro studies using VERO and MDCK cell lineages were performed to study the cytotoxicity and cell uptake of the SPIONs. For both cell lineages, PEG- and DEXTRAN-coated nanoparticles presented high cell viability for concentrations as high as 200 µg/mL. In vivo studies were conducted using BALB/c mice inoculating the SPIONs intravenously and exposing them to the presence of an external magnet located over the tumour. It was observed that the amount of PEG-coated SPIONs in the tumor increased by up to 160% when using the external permanent magnetic as opposed to those animals that were not exposed to the external magnetic field.
Subject(s)
Dextrans/pharmacokinetics , Ferric Compounds/pharmacokinetics , Magnetic Fields , Nanoparticles , Animals , Chlorocebus aethiops , Dextrans/administration & dosage , Dextrans/toxicity , Dogs , Drug Carriers , Drug Evaluation, Preclinical , Female , Ferric Compounds/administration & dosage , Ferric Compounds/toxicity , In Vitro Techniques , Injections, Intravenous , Liver/metabolism , Lung/metabolism , Madin Darby Canine Kidney Cells , Magnetite Nanoparticles/administration & dosage , Magnetite Nanoparticles/toxicity , Mammary Neoplasms, Experimental/metabolism , Materials Testing , Mice , Mice, Inbred BALB C , Nanoparticles/administration & dosage , Nanoparticles/toxicity , Polyethylene Glycols , Skin/metabolism , Spectroscopy, Fourier Transform Infrared , Tissue Distribution , Vero CellsABSTRACT
Magnetic hyperthermia (MH) is based on the use of magnetic nanoparticles (MNPs) to selectively increase the temperature of MNP-loaded target tissues when applying an alternating magnetic field (AMF) in the range of radiofrequency. To date, all MH research has focused on heat generation in an attempt to elucidate the mechanisms for the death of MNP-loaded cells submitted to AMF. However, recent in vitro studies have demonstrated the feasibility of inducing dramatic cell death without increasing the macroscopic temperature during AMF exposure. Here, we show that the cell death observed following AMF exposure, specifically that of MNP-loaded dendritic cells (DCs) in culture, was caused by the release of toxic agents into the cell culture supernatants and not due to a macroscopic temperature increase. We performed MH in vitro experiments to demonstrate that the supernatant of the cell culture following AMF exposure was highly toxic when added to control unloaded DCs, as this treatment led to nearly 100% cell death. Therefore, our results demonstrate that heat is not the only agent responsible for triggering cell death following MH treatment. This finding offers new perspectives for the use of DCs as the proverbial Trojan horse to vectorise MNPs to the target tumour area and these results further support the use of DCs as therapeutic agents against cancer when submitted to AMF. Furthermore, this discovery may help in understanding the mechanism of cell death mediated by exposure to AMF.
Subject(s)
Apoptosis , Dendritic Cells/cytology , Hyperthermia, Induced , Magnetite Nanoparticles/chemistry , Cells, Cultured , Dendritic Cells/drug effects , Humans , Magnetic Fields , Magnetite Nanoparticles/therapeutic use , Magnetite Nanoparticles/toxicity , Neoplasms/drug therapy , Rhodamines/chemistryABSTRACT
BACKGROUND: Magnetic hyperthermia is currently a clinical therapy approved in the European Union for treatment of tumor cells, and uses magnetic nanoparticles (MNPs) under time-varying magnetic fields (TVMFs). The same basic principle seems promising against trypanosomatids causing Chagas disease and sleeping sickness, given that the therapeutic drugs available have severe side effects and that there are drug-resistant strains. However, no applications of this strategy against protozoan-induced diseases have been reported so far. In the present study, Crithidia fasciculata, a widely used model for therapeutic strategies against pathogenic trypanosomatids, was targeted with Fe(3)O(4) MNPs in order to provoke cell death remotely using TVMFs. METHODS: Iron oxide MNPs with average diameters of approximately 30 nm were synthesized by precipitation of FeSO(4) in basic medium. The MNPs were added to C. fasciculata choanomastigotes in the exponential phase and incubated overnight, removing excess MNPs using a DEAE-cellulose resin column. The amount of MNPs uploaded per cell was determined by magnetic measurement. The cells bearing MNPs were submitted to TVMFs using a homemade AC field applicator (f = 249 kHz, H = 13 kA/m), and the temperature variation during the experiments was measured. Scanning electron microscopy was used to assess morphological changes after the TVMF experiments. Cell viability was analyzed using an MTT colorimetric assay and flow cytometry. RESULTS: MNPs were incorporated into the cells, with no noticeable cytotoxicity. When a TVMF was applied to cells bearing MNPs, massive cell death was induced via a nonapoptotic mechanism. No effects were observed by applying TVMF to control cells not loaded with MNPs. No macroscopic rise in temperature was observed in the extracellular medium during the experiments. CONCLUSION: As a proof of principle, these data indicate that intracellular hyperthermia is a suitable technology to induce death of protozoan parasites bearing MNPs. These findings expand the possibilities for new therapeutic strategies combating parasitic infection.
Subject(s)
Crithidia fasciculata/physiology , Crithidia fasciculata/radiation effects , Euglenozoa Infections/parasitology , Euglenozoa Infections/therapy , Hyperthermia, Induced/methods , Magnetic Field Therapy/methods , Magnetite Nanoparticles/therapeutic use , Animals , Cells, Cultured , Humans , Treatment OutcomeABSTRACT
PURPOSE: To investigate the effects of alternating magnetic fields (AMF) on the death rate of dendritic cells (DCs) loaded with magnetic nanoparticles (MNPs) as heating agents. AMF exposure time and amplitude as well as the MNPs concentration were screened to assess the best conditions for a controlled field-induced cell death. METHODS: Human-monocyte-derived DCs were co-incubated with dextran-coated MNPs. The cells were exposed to AMF (f = 260 kHz; 0 < H(0) < 12.7 kA/m) for intervals from 5 to 15 min. Morphology changes were assessed by scanning electron microscopy. Cell viability was measured by Trypan blue and fluorescence-activated cell sorting (FACS) using Annexin-propidium iodide markers. RESULTS: We were able to control the DCs viability by a proper choice AMF amplitude and exposure time, depending on the amount of MNPs uploaded. About 20% of cells showed Annexin-negative/PI-positive staining after 5-10 min of AMF exposure. CONCLUSIONS: Controlled cell death of MNP-loaded DCs can be obtained by adequate tuning of the physical AMF parameters and MNPs concentration. Necrotic-like populations were observed after exposure times as short as 10 min, suggesting a fast underlying mechanism for cell death. Power absorption by the MNPs might locally disrupt endosomic membranes, thus provoking irreversible cell damage.
Subject(s)
Cell Death , Hyperthermia, Induced , Magnetics , Metal Nanoparticles , Cell Survival , Dendritic Cells/cytology , Dendritic Cells/metabolism , Dendritic Cells/pathology , Ferric Compounds/metabolism , Flow Cytometry , Humans , Magnetic Fields , Microscopy, Electron, Scanning , Time FactorsABSTRACT
In this work, the capability of primary, monocyte-derived dendritic cells (DCs) to uptake iron oxide magnetic nanoparticles (MNPs) is assessed and a strategy to induce selective cell death in these MNP-loaded DCs using external alternating magnetic fields (AMFs) is reported. No significant decrease in the cell viability of MNP-loaded DCs, compared to the control samples, was observed after five days of culture. The number of MNPs incorporated into the cytoplasm was measured by magnetometry, which confirmed that 1-5 pg of the particles were uploaded per cell. The intracellular distribution of these MNPs, assessed by transmission electron microscopy, was found to be primarily inside the endosomic structures. These cells were then subjected to an AMF for 30 min and the viability of the blank DCs (i.e. without MNPs), which were used as control samples, remained essentially unaffected. However, a remarkable decrease of viability from approximately 90% to 2-5% of DCs previously loaded with MNPs was observed after the same 30 min exposure to an AMF. The same results were obtained using MNPs having either positive (NH(2)(+)) or negative (COOH(-)) surface functional groups. In spite of the massive cell death induced by application of AMF to MNP-loaded DCs, the number of incorporated magnetic particles did not raise the temperature of the cell culture. Clear morphological changes at the cell structure after magnetic field application were observed using scanning electron microscopy. Therefore, local damage produced by the MNPs could be the main mechanism for the selective cell death of MNP-loaded DCs under an AMF. Based on the ability of these cells to evade the reticuloendothelial system, these complexes combined with an AMF should be considered as a potentially powerful tool for tumour therapy.
Subject(s)
Dendritic Cells/cytology , Magnetics/methods , Nanoparticles/chemistry , Absorption , Antigens, Surface/metabolism , Cell Death , Cell Differentiation , Cell Survival , Cells, Cultured , Colloids , Dendritic Cells/ultrastructure , Endocytosis , Flow Cytometry , Humans , Hydrodynamics , Leukocytes, Mononuclear/cytology , Light , Nanoparticles/ultrastructure , Particle Size , Scattering, Radiation , Temperature , Trypan Blue/metabolismABSTRACT
We report on novel ferrogels derived from polysaccharides (sodium alginate and chitosan) with embedded iron oxide nanoparticles synthesized in situ and their combination with thermally responsive poly(N-isopropylacrylamide) for externally driven drug release using AC magnetic fields. Samples were characterized by Raman spectroscopy, transmission electron microscopy, and magnetic measurements. The obtained nanoparticles were found to be of â¼10 nm average size, showing magnetic properties very close to those of the bulk material. The thermal response was measured by power absorption experiments, finding specific power absorption values between 100 and 300 W/g, which was enough for attaining the lower critical solution temperature of the polymeric matrix within few minutes. This fast response makes these materials good candidates for externally controlled drug release.
Subject(s)
Alginates/chemistry , Chitosan/chemistry , Drug Carriers/chemistry , Hydrogels/chemistry , Magnetics , Absorption , Ferric Compounds/chemistry , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Hot Temperature , Nanoparticles/chemistryABSTRACT
Spherical shaped nanoparticles of series Y(2 - x)Eu(x)O(3) (x = 0.06, 0.10, 0.20, and 2) and Gd(2 - x)Eu(x)O(3) (x = 0.06, 0.10) were prepared by thermolysis of 2,4-pentanedione complexes of Y, Gd, and Eu. The bixbyite phase of Gd(2 - x)Eu(x)O(3) samples was formed at 500 degrees C, whereas the thermal decomposition of Y and Eu complexes' mixtures occurred at higher temperatures. Linearity in the concentration dependence on lattice parameter confirmed the formation of solid solutions. The distribution of Eu(3+) in Gd(2 - x)Eu(x)O(3) was changed with thermal annealing: in the as-prepared sample (x = 0.10) the distribution was preferential at C(3i) sites while in the annealed samples, Eu(3+) were distributed at both C(2) and C(3i) sites. Rietveld refinement of site occupancies as well as emission spectra showed a random distribution of cations in Y(2 - x)Eu(x)O(3). The photoluminescence (PL) measurements of the sample showed red emission with the main peak at 614 nm ((5)D(0)-(7)F(2)). The PL intensity increased with increasing concentration of Eu(3+) in both series. Infrared excitation was required to obtain good Raman spectra. The linear dependence of the main Raman peak wavenumber offers a non-destructive method for monitoring the substitution level and its homogeneity at the micron scale.
ABSTRACT
We have investigated the internalization of magnetic nanoparticles (NPs) into dendritic cells (DCs) in order to assess both the final location of the particles and the viability of the cultured cells. The particles, consisting of a metallic iron core covered with carbon, showed no toxic effects on the DCs and had no effect in their viability. We found that mature DCs are able to incorporate magnetic nanoparticles in a range of size from 10 nm to ca. 200 nm, after 24 h of incubation. We describe a method to separate cells loaded with NPs, and analyze the resulting material by electron microscopy and magnetic measurements. It is found that NPs are internalized in lysosomes, providing a large magnetic signal. Our results suggest that loading DCs with properly functionalized magnetic NPs could be a promising strategy for improved vectorization in cancer diagnosis and treatment.
Subject(s)
Cell Separation/methods , Dendritic Cells/metabolism , Magnetics , Metal Nanoparticles , Dendritic Cells/ultrastructure , Humans , Lysosomes/metabolism , Microscopy, Electron, TransmissionABSTRACT
There has been no consistent approach to the follow-up of Kawasaki disease patients for remote coronary perfusion abnormalities. Dobutamine stress echocardiography (DSE) has become a standard method for evaluation of perfusion abnormalities in adults with coronary artery disease. In addition, DSE has been used with success in some pediatric patients. The purposes of this study were to evaluate safety and accuracy of DSE in the follow-up of patients with Kawasaki disease, to evaluate whether DSE adds any additional value to the resting echocardiogram, and to determine the association of DSE results with American Heart Association (AHA) risk level categories. DSE was performed 1 month to 13 years after acute Kawasaki disease in 47 patients (range, 3.8-22.6 years; 33 males and 16 females). Patients were stratified according to AHA risk level categories (I-V). Ischemia was defined as a new or worsening regional wall motion abnormality or >1 mm ST segment depression on the electrocardiogram during DSE. In 45/47 patients, DSE was completed successfully (i.e., achievement of target heart rate or development of ischemia). No patients in risk levels lower than V (i.e., patients without coronary artery stenoses) had positive DSE, whereas 2/4 (50%) in the risk level V category had positive DSE, both of whom had coronary occlusion >50% confirmed by angiography. Of the 2 AHA risk level V patients with negative DSE, 1 had extensive collateralization and the other had coronary obstruction <50%. DSE is a safe and feasible method for the evaluation of children with Kawasaki disease. DSE provides a confirmatory benefit and may be a useful screening alternative to cardiac catheterization during follow-up. Patients in AHA risk levels I-IV are unlikely to have dobutamine-induced coronary perfusion abnormalities. Patients in the risk level V category may or may not have positive DSE depending on the degree of both coronary obstruction and collateralization.