Síntese e caracterização de nanopartículas magnéticas de óxido de ferro para aplicações biomédicas – um estudo citotóxico em linhagem celular de carcinoma cervical humano (células HeLa)

Nanopartículas magnéticas (NPMs) têm sido alvo de inúmeras investigações por seu grande potencial de aplicação nos mais diferentes campos tecnológicos. Dentre tantos, elas destacam-se na área biomédica, seja no diagnóstico ou tratamento de diversas doenças. Neste trabalho foram sintetizadas nanopartículas magnéticas (NPMs) de óxido de ferro (magnetita) pelo método de coprecipitação de íons Fe2+ e Fe3+ em meio alcalino. O objetivo central foi estudar as características morfológicas, estruturais, magnéticas e o comportamento biológico desses compostos em células cancerígenas, visando futuras aplicações biomédicas. Inicialmente as nanopartículas magnéticas foram avaliadas em função dos parâmetros físico-químicos que influenciam diretamente as características finais do produto (pH, molaridade, temperatura e tipo de base) para se observar as melhores condições de síntese e a influência de cada um nas características do produto. As nanopartículas foram caracterizadas por difratometria de raios-X, microscopia eletrônica de varredura, potencial zeta e magnetometria. Desse primeiro estudo concluiu-se que a coprecipitação produz partículas com polidispersão de tamanhos alta e que os parâmetros de síntese influenciam drasticamente as propriedades dos materiais, no entanto, todas as amostras exibiam características magnéticas. Depois de estabelecida esta etapa, as NPMs foram submetidas à transfecção em cultura celular de carcinoma cervical humano (células HeLa) e a testes biológicos como coloração com Azul da Prússia e hematoxilina-eosina, ensaio de MTT e ensaio de apoptose para averiguação da citotoxidade. A principal observação vinda desses resultados foi que as nanopartículas magnéticas sintetizadas, salvo algumas adaptações de síntese...Magnetic nanoparticles (MNPs) have been the subject of numerous investigations because of its great potential application in many different fields of technology. Among many, they stand out in the biomedical area, either in diagnosis or treatment of various diseases. In this work were synthesized magnetic nanoparticles (MNPs) of iron oxide (magnetite) by the coprecipitation method of Fe2+ and Fe3+ in an alkaline medium. The main objective was to study the morphological, structural, magnetic and biological behavior of these compounds in cancer cells, in order to future biomedical applications. Initially the magnetic nanoparticles were evaluated against the physical and chemical parameters that directly influence the final characteristics of product (pH, molarity, temperature and type of base) to observe the best synthesis conditions and influence of each characteristics. The nanoparticles were characterized by X-ray diffraction, sccaning electron microscopy, zeta potential and magnetometry. In this first study showed that coprecipitation produces particles with high polydispersity of sizes and that the synthesis parameters dramatically influence the properties of materials, however all samples exhibited magnetic characteristics. After you make this step, the MNP were subjected to transfection in cell culture of human cervical carcinoma (HeLa) and biological tests such as staining with Prussian blue and hematoxylin-eosin, MTT assay and apoptosis assay to investigate cytotoxicity. The main observation was that these results coming magnetic nanoparticles synthesized, except for some adjustments in short, constitute a class of nanocomposites with enormous potential for therapeutic and / or diagnosis. The work brings together information ranging from the synthesis of nanoparticles to their behavior inside the cells, emphasizing the best conditions for each procedureConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Hematite Surface Modification toward Efficient Sunlight-Driven Water Splitting Activity : The Role of Gold Nanoparticle Addition

Localized surface plasmon resonance has been investigated to enhance light harvesting in hematite-based photoelectrodes modified with gold nanoparticles (AuNPs); meanwhile, an extensive understanding about the different processes involved in the hematite-AuNP system remains unclear. This work addresses a majority of effects associated with AuNP addition by comparing charge transfer, catalytic and light harvesting efficiencies. The obtained results revealed that the lower AuNP amount leads to a higher photocurrent response of 1.20 mA cm-2 at 1.23 VRHE in comparison with all photoelectrodes designed here. X-ray photoelectron data revealed that hematite photoelectrodes loaded with higher concentrations of AuNPs immersed in an alkaline electrolyte showed hydrated/oxidized gold phase formation at the electrode/electrolyte interface. This change on the semiconductor-metal interface may affect the conductivity impairing the photocatalytic performance because of the passivation layer on the AuNP surface, decreasing the efficiency of charge transfer. Notoriously, increasing AuNP amount supported on the hematite surface clearly promoted higher light absorption, which was surprisingly not followed by photoelectrochemical efficiency. This result suggests here that the plasmon effect is not a dominant phenomenon that drives the photoelectrode performance. In fact, a deeper analysis showed that the loaded hematite photoelectrodes with low amounts of AuNPs provides a Schottky contact at the semiconductor-metal interface leading to Fermi level equilibration enhancing charge transport efficiency, which is classified as the predominant effect leading to higher photoresponse in the system.acceptedVersionPeer reviewe

Biological Oxidative Mechanisms for Degradation of Poly(lactic acid) Blended with Thermoplastic Starch

In the present study, poly­(lactic acid) (PLA) and their blends with 5%/wt and 10%/wt thermoplastic starch (TPS) were submitted to degradation in simulated soil. To investigate the mechanisms involved in the degradation, we also submitted the samples to degradation by <i>tert</i>-butyl hydroperoxide, myoglobin, and peroxide-activated myoglobin. The samples were analyzed by Fourier-transformed infrared spectrometry (FTIR), scanning electronic microscopy (SEM), contact angle analysis, and mass loss measurement. The FTIR results indicated a weak interaction between the two components (PLA and starch) in the blend’s amorphous structure. However, the corresponding SEM images showed that TPA increased ridges and roughness at the material surface associated with an increase of wettability evidenced by contact angle analysis. Consistently, TPS favored degradation of the material both in the simulated soil and pro-oxidant model systems. In the simulated soil, the occurrence of TPS hydrolysis provided glucose, a biological fuel, that contributed to the growth of the microorganisms. The similar degradation patterns observed in mimetic pro-oxidant biological systems and soil suggest that oxidative reactions catalyzed by heme proteins from biological sources as well as the presence of peroxides and transition metal traces in the original materials have a significant contribution to PLA and PLA/TPS degradation

Variable density and viscosity, miscible displacements in horizontal Hele-Shaw cells. Part 2. Nonlinear simulations

Magnetic iron oxide nanoparticles (magnetite) (MNPs) were prepared using different organic and inorganic bases. Strong inorganic base (KOH) and organic bases (NH4OH and 1,4-diazabicyclo[2.2.2]octane (DABCO)) were used in the syntheses of the MNPs. The MNPs were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM). Fourier transform infrared spectroscopy (FT-IR) and magnetization measurements. MNPs prepared with strong inorganic base yielded an average size of 100 nm, whereas the average size of the MNPs prepared with the organic bases was 150 nm. The main competitive phase for MNPs prepared with the strong inorganic and organic bases was maghemite; however, syntheses with KOH yielded a pure magnetite phase. The transfection study performed with the MNPs revealed that the highest transfection rate was obtained with the MNPs prepared with KOH (74%). The correlation between the magnetic parameters and the transfection ratio without transfection agents indicated that MNPs prepared with KOH were a better vector for possible applications of these MNPs in biomedicine. HeLa cells incubated with MNP-KOH at 10 mu g/mL for 24 and 48 h exhibited a decrease in population in comparison with the control cells and it was presumably related to the toxicity of the MNPs. However, the cells incubated with MNP-KOH at 50 and 100 mu g/mL presented a very small difference in the viability between the cell populations studied at 24 and 48 h. These data illustrate the viability of HeLa cells treated with MNP-KOH and suggest the potential use of these MNPs in biomedical applications. (C) 2012 Elsevier B.V. All rights reserved.Sao Paulo Science Foundation (FAPESP) [05/54703-6, 09/11203-4]Sao Paulo Science Foundation (FAPESP)National Council for Scientific and Technological Development (CNPq)National Council for Scientific and Technological Development (CNPq) [301596/2011-5]Braile Biomedica Industria, Comercio e Representacoes LimitadaBraile Biomedica Industria, Comercio e Representacoes Limitad

pH-Dependent Synthesis of Anisotropic Gold Nanostructures by Bioinspired Cysteine-Containing Peptides

In the present study, alkaline peptides AAAXCX (X = lysine or arginine residues) were designed based on the conserved motif of the enzyme thioredoxin and used for the synthesis of gold nanoparticles (GNPs) in the pH range of 2–11. These peptides were compared with free cysteine, the counterpart acidic peptides AAAECE and γ-ECG (glutathione), and the neutral peptide AAAACA. The objective was to investigate the effect of the amino acids neighboring a cysteine residue on the pH-dependent synthesis of gold nanocrystals. Kohn–Sham density functional theory (KS-DFT) calculations indicated an increase in the reducing capacity of AAAKCK favored by the successive deprotonation of their ionizable groups at increasing pH values. Experimentally, it was observed that gold speciation and the peptide structure also have a strong influence on the synthesis and stabilization of GNPs. AAAKCK produced GNPs at room temperature, in the whole investigated pH range. By contrast, alkaline pH was the best condition for the synthesis of GNP assisted by the AAARCR peptide. The acidic peptides produced GNPs only in the presence of polyethylene glycol, and the synthesis using AAAECE and γ-ECG also required heating. The ionization state of AAAKCK had a strong influence on the preferential growth of the GNPs. Therefore, pH had a remarkable effect on the synthesis, kinetics, size, shape, and polydispersity of GNPs produced using AAAKCK. The AAAKCK peptide produced anisotropic decahedral and platelike nanocrystals at acidic pH values and spherical GNPs at alkaline pH values. Both alkaline peptides were also efficient capping agents for GNPs, but they produced a significant difference in the zeta potential, probably because of different orientations on the gold surface

Intermediate Tyrosyl Radical and Amyloid Structure in Peroxide-Activated Cytoglobin.

We characterized the peroxidase mechanism of recombinant rat brain cytoglobin (Cygb) challenged by hydrogen peroxide, tert-butylhydroperoxide and by cumene hydroperoxide. The peroxidase mechanism of Cygb is similar to that of myoglobin. Cygb challenged by hydrogen peroxide is converted to a Fe4+ oxoferryl π cation, which is converted to Fe4+ oxoferryl and tyrosyl radical detected by direct continuous wave-electron paramagnetic resonance and by 3,5-dibromo-4-nitrosobenzene sulfonate spin trapping. When organic peroxides are used as substrates at initial reaction times, and given an excess of peroxide present, the EPR signals of the corresponding peroxyl radicals precede those of the direct tyrosyl radical. This result is consistent with the use of peroxide as a reducing agent for the recycling of Cygb high-valence species. Furthermore, we found that the Cygb oxidation by peroxides leads to the formation of amyloid fibrils. This result suggests that Cygb possibly participates in the development of degenerative diseases; our findings also support the possible biological role of Cygb related to peroxidase activity

Interatoma of rat Cygb with hydrogen peroxide.

<p>The network shows, in each node, a protein predicted to have functional links with Cygb and hydrogen peroxide. Inside the figure the abbreviations are SOD1 (superoxide dismutase [Cu-Zn]), Hmox2 (heme oxygenase 2 [HO-2]), Mb (myoglobin), Mpo (myeloperoxidase), cat (catalase), Cygb (cytoglobin), Prdx1 (peroxyredoxin-1), Prdx5 (peroxyredoxin-5) and Srxn1 (Ab2-390). In the figure light green, cyan and magenta lines correspond, respectively, to textmining, databases and experiments supporting the relationship among the proteins and hydrogen peroxide.</p

Changes in the EA spectrum of Cygb during the reaction with hydrogen peroxide.

<p>A) Bleaching of Soret and Q bands of EA spectra of Cygb in the course of the reaction with hydrogen peroxide. The black line represents the EA spectrum of resting Cygb, red, green and blue lines corresponds to the spectra obtained at 30, 60 and 200 s after addition of hydrogen peroxide and indicated by the arrows. B) Normalized spectra of Cygb resting form and 200 s after hydrogen peroxide addition. C) Differential spectra of Cygb obtained 30 and 200 s after the addition of hydrogen peroxide The experiments of EA spectroscopy were performed using 65 μmol.L^-1 Cygb and 0.1 cm optical length. When present, the concentration of peroxide was 650 μmol.L^-1. These results are representative of three independent replicates.</p

Formation of Cygb amyloid structure after challenge by peroxides.

<p>A), B) and C) show, respectively the epifluorescence images of Cygb control, control plus GSH and challenged by hydrogen peroxide obtained immediately (left panels) 24 h (right panels) after incubation and staining by thioflavine-T. For the low-vacuum SEM experiments, it was used 7 μmol.L^-1 cygb solution with 70 μmol.L^-1 peroxide solutions. For the epifluorescence experiments 70 μmol.L^-1 protein solution was incubated for 1 h with 700 μmol.L^-1 peroxide solution in the presence of thioflavin-T. For FTIR measurements, 7 μmol.L^-1 protein solution was incubated with 70 μmol.L^-1 peroxide solutions for 1 h. The results are representative of three independent experiments.</p