Electro-optics of confined systems.

Confinement in microenvironments occurs in many natural systems and technological applications. However, little is known about the behaviour of the immersed nanoparticles. In this work we show that their diffusion, electro-orientation and electric field induced polarization can be determined through electric birefringence experiments. We analyze aqueous dispersions of silver nanowires and clay particles confined inside microdroplets. We have observed that confinement reduces the amount of particles that can be oriented by the external electric field. However, the polarizability of the oriented particles is not affected by the presence of the oil/water boundary, and it is the same as in unbounded media, which agrees with the fact that the electric polarization and related phenomena are short-ranged.

Effects of layer-by-layer coating on activated carbon electrodes for capacitive deionization.

Recently, the need for obtaining, reusing, or purifying water has become a crucial issue. The capacitive deionization (CDI) method, which is based on the electric double layer (EDL) concept, can be applied to ion adsorption from an aqueous solution. This process is carried out by applying a potential difference to highly porous electrodes while pumping salty solution between them, partially removing the ions present in the solution and keeping them in the surface of the electrodes. The use of coated carbon electrodes with one polyelectrolyte layer, turning them into "soft electrodes" (SEs), has been proved to improve the efficiency of the system with respect to its original configuration. In this work, we investigate the effect on the ion adsorption and the efficiency of the process when implementing the coating technique known as layer-by-layer (LbL) on the electrode. This consists in successively coating the electrode surfaces with polyelectrolyte layers, alternating their charge polarity in each step. We tested the effect of the number of layers deposited, as well as the impact of this technique by using different carbons. We found that the second polyelectrolyte layer adheres more than the first layer, serving as a support or seed when it is not dense and uniformly distributed. In contrast, if the first layer is well adhered, a third layer is needed to observe improvements in adsorption and process efficiency. The adsorption of the polymer layers depends in any instance on the porosity of the carbon.

Electro-optical Study of the Anomalous Rotational Diffusion in Polymer Solutions.

Brownian diffusion of spherical nanoparticles is usually exploited to ascertain the rheological properties of complex media. However, the behavior of the tracer particles is affected by a number of phenomena linked to the interplay between the dynamics of the particles and polymer coils. For this reason, the characteristic lengths of the dispersed entities, depletion phenomena, and the presence of sticking conditions have been observed to affect the translational diffusion of the probes. On the other hand, the retardation effect of the host fluid on the rotational diffusion of nonspherical particles is less understood. We explore the possibility of studying this phenomenon by analyzing the electro-orientation of the particles in different scenarios in which we vary the ratio between the particle and polymer characteristic size, and the geometry of the particles, including both elongated and oblate shapes. We find that the Stokes-Einstein relation only applies if the radius of gyration of the polymer is much shorter than the particle size and when some repulsive interaction between both is present.

Combined Magnetic Hyperthermia and Photothermia with Polyelectrolyte/Gold-Coated Magnetic Nanorods.

Magnetite nanorods (MNRs) are synthesized based on the use of hematite nanoparticles of the desired geometry and dimensions as templates. The nanorods are shown to be highly monodisperse, with a 5:1 axial ratio, and with a 275 nm long semiaxis. The MNRs are intended to be employed as magnetic hyperthermia and photothermia agents, and as drug vehicles. To achieve a better control of their photothermia response, the particles are coated with a layer of gold, after applying a branched polyethyleneimine (PEI, 2 kDa molecular weight) shell. Magnetic hyperthermia is performed by application of alternating magnetic fields with frequencies in the range 118-210 kHz and amplitudes up to 22 kA/m. Photothermia is carried out by subjecting the particles to a near-infrared (850 nm) laser, and three monochromatic lasers in the visible spectrum with wavelengths 480 nm, 505 nm, and 638 nm. Best results are obtained with the 505 nm laser, because of the proximity between this wavelength and that of the plasmon resonance. A so-called dual therapy is also tested, and the heating of the samples is found to be faster than with either method separately, so the strengths of the individual fields can be reduced. Due to toxicity concerns with PEI coatings, viability of human hepatoblastoma HepG2 cells was tested after contact with nanorod suspensions up to 500 µg/mL in concentration. It was found that the cell viability was indistinguishable from control systems, so the particles can be considered non-cytotoxic in vitro. Finally, the release of the antitumor drug doxorubicin is investigated for the first time in the presence of the two external fields, and of their combination, with a clear improvement in the rate of drug release in the latter case.

Electro-orientation of Ag nanowires in viscoelastic fluids.

In this work we study the electro-orientation (through electric birefringence experiments) of silver nanowires in polymer solutions eventually capable of forming gel networks. Information on the structure of the polymer solution is obtained by evaluating the electro-orientation of the nanowires. It is found that in presence of poly(ethylene oxide), Kerr's law (birefringence proportional to the square of the field) is fulfilled, and the randomization process after switching off the external field is purely diffusive, controlled by the viscosity of the Newtonian polymer solution. In the case of (gelating) sodium alginate solutions, measuring at larger distances from the bottom (where the source of cross-linking Ca2+ ions is deposited) means a smaller degree of cross-linking, and a less stiff gel. In fact, it is found that after a certain time the birefringence signal gets frozen at the bottom, indicating that a gel network is formed which hinders particle orientation. The viscosity deduced up to that point agrees well with rheological determinations, with increasing deviations found at longer times due to the inhomogeneous gel formation. This process has an interesting consequence on birefringence response: Kerr's law fails to be fulfilled, appearing a "yield" applied electric field, larger the longer the time after preparation.

Poly(ethylene-imine)-Functionalized Magnetite Nanoparticles Derivatized with Folic Acid: Heating and Targeting Properties.

Magnetite nanoparticles (MNPs) coated by branched poly (ethylene-imine) (PEI) were synthesized in a one-pot. Three molecular weights of PEI were tested, namely, 1.8 kDa (sample MNP-1), 10 kDa (sample MNP-2), and 25 kDa (sample MNP-3). The MNP-1 particles were further functionalized with folic acid (FA) (sample MNP-4). The four types of particles were found to behave magnetically as superparamagnetic, with MNP-1 showing the highest magnetization saturation. The particles were evaluated as possible hyperthermia agents by subjecting them to magnetic fields of 12 kA/m strength and frequencies ranging between 115 and 175 kHz. MNP-1 released the maximum heating power, reaching 330 W/g at the highest frequency, in the high side of reported values for spherical MNPs. In vitro cell viability assays of MNP-1 and MNP-4 against three cell lines expressing different levels of FA receptors (FR), namely, HEK (low expression), and HeLa (high expression), and HepG2 (high expression), demonstrated that they are not cytotoxic. When the cells were incubated in the presence of a 175 kHz magnetic field, a significant reduction in cell viability and clone formation was obtained for the high expressing FR cells incubated with MNP-4, suggesting that MNP-4 particles are good candidates for magnetic field hyperthermia and active targeting.

Modulation of the Magnetic Hyperthermia Response Using Different Superparamagnetic Iron Oxide Nanoparticle Morphologies.

The use of magnetic nanoparticles in hyperthermia, that is, heating induced by alternating magnetic fields, is gaining interest as a non-invasive, free of side effects technique that can be considered as a co-adjuvant of other cancer treatments. Having sufficient control on the field characteristics, within admissible limits, the focus is presently on the magnetic material. In the present contribution, no attempt has been made of using other composition than superparamagnetic iron oxide nanoparticles (SPION), or of applying surface functionalization, which opens a wider range of choices. We have used a hydrothermal synthesis route that allows preparing SPION nanoparticles in the 40 nm size range, with spherical, cuboidal or rod-like shapes, by minor changes in the synthesis steps. The three kinds of particles (an attempt to produce star-shaped colloids yielded hematite) were demonstrated to have the magnetite (or maghemite) crystallinity. Magnetization cycles showed virtually no hysteresis and demonstrated the superparamagnetic nature of the particles, cuboidal ones displaying saturation magnetization comparable to bulk magnetite, followed by rods and spheres. The three types were used as hyperthermia agents using magnetic fields of 20 kA/m amplitude and frequency in the range 136-205 kHz. All samples demonstrated to be able to raise the solution temperature from room values to 45 °C in a mere 60 s. Not all of them performed the same way, though. Cuboidal magnetic nanoparticles (MNPs) displayed the maximum heating power (SAR or specific absorption rate), ranging in fact among the highest reported with these geometries and raw magnetite composition.

Polymer-induced orientation of nanowires under electric fields.

The controlled orientation of metallic wires inside a polymeric medium can enhance desired properties of the composites, such as the electrical conductivity or the optical transmittance. In this work, we study silver nanowire orientation in semidilute suspensions of DNA and find an intriguing effect: under the application of low-frequency AC electric fields with moderate amplitude, the DNA coils can provoke the orientation of the wires in solution. The phenomenon is entirely induced by the polymer, when it is deformed by the application of an electric field. This effect is explained using computer simulations based on excluded-volume interactions. Moreover, we experimentally show that such a behaviour is not exclusive of silver nanowire-DNA suspensions, but rather occurs for other particle-polymer systems. This phenomenon can be taken advantage of to achieve strong orientation of particles otherwise insensitive to electric fields.

Gemcitabine-Loaded Magnetically Responsive Poly(ε-caprolactone) Nanoparticles against Breast Cancer.

A reproducible and efficient interfacial polymer disposition method has been used to formulate magnetite/poly(ε-caprolactone) (core/shell) nanoparticles (average size ≈ 125 nm, production performance ≈ 90%). To demonstrate that the iron oxide nuclei were satisfactorily embedded within the polymeric solid matrix, a complete analysis of these nanocomposites by, e.g., electron microscopy visualizations, energy dispersive X-ray spectroscopy, Fourier-transform infrared spectroscopy, electrophoresis, and contact angle goniometry was conducted. The magnetic responsive behaviour of these nanoparticles was quantitatively characterized by the hysteresis cycle and qualitatively investigated by visualization of the colloid under exposure to a 0.4 T magnet. Gemcitabine entrapment into the polymeric shell reported adequate drug loading values (≈11%), and a biphasic and pH-responsive drug release profile (≈ four-fold faster Gemcitabine release at pH 5.0 compared to pH 7.4). Cytotoxicity studies in MCF-7 human breast cancer cells proved that the half maximal inhibitory concentration of Gem-loaded nanocomposites was ≈ two-fold less than that of the free drug. Therefore, these core/shell nanoparticles could have great possibilities as a magnetically targeted Gemcitabine delivery system for breast cancer treatment.

AC Electrokinetics of Salt-Free Multilayered Polymer-Grafted Particles.

Interest in the electrical properties of the interface between soft (or polymer-grafted) nanoparticles and solutions is considerable. Of particular significance is the case of polyelectrolyte-coated particles, mainly taking into account that the layer-by-layer procedure allows the control of the thickness and permeability of the layer, and the overall charge of the coated particle. Like in simpler systems, electrokinetic determinations in AC fields (including dielectric dispersion in the 1 kHz-1 MHz frequency range and dynamic electrophoresis by electroacoustic methods in the 1-18 MHz range) provide a large amount of information about the physics of the interface. Different models have dealt with the electrokinetics of particles coated by a single polymer layer, but studies regarding multi-layered particles are far scarcer. This is even more significant in the case of so-called salt-free systems; ideally, the only charges existing in this case consist of the charge in the layer(s) and the core particle itself, and their corresponding countercharges, with no other ions added. The aims of this paper are as follows: (i) the elaboration of a model for the evaluation of the electrokinetics of multi-grafted polymer particles in the presence of alternating electric fields, in dispersion media where no salts are added; (ii) to carry out an experimental evaluation of the frequency dependence of the dynamic (or AC) electrophoretic mobility and the dielectric permittivity of suspensions of polystyrene latex spherical particles coated with successive layers of cationic, anionic, and neutral polymers; and (iii) finally, to perform a comparison between predictions and experimental results, so that it can be demonstrated that the electrokinetic analysis is a useful tool for the in situ characterization of multilayered particles.

First steps in the formulation of praziquantel nanosuspensions for pharmaceutical applications.

Praziquantel (PZQ), a broad spectrum anthelmintic drug, cannot be found in acceptable dosage forms for elderly patients, paediatric patients, and for veterinary use. In fact, very little has been done up to now in the formulation of liquid dosage forms, being they always formulated for parenteral administration. To beat this important challenge, it was accomplished a comprehensive analysis of the influence of two elementary physicochemical aspects, i.e. surface thermodynamic and electrokinetic properties, on the colloidal stability of PZQ nanosuspensions. The hydrophobic character of the drug, intensely determining the flocculation curves, was confirmed by the thermodynamic characterization. The electrophoretic characterization, in combination with the sedimentation and relative absorbance versus time curves, highlighted that the electrical double layer thickness and the surface charge can play an essential role in the stability of the pharmaceutical colloid. Finally, it was demonstrated that controlling the pH values and the incorporation of electrolytes can help in formulating PZQ aqueous nanosuspensions with appropriate stability and redispersibility behaviours for pharmaceutical use.

Hyperthermia-Triggered Doxorubicin Release from Polymer-Coated Magnetic Nanorods.

In this paper, it is proposed that polymer-coated magnetic nanorods (MNRs) can be used with the advantage of a double objective: first, to serve as magnetic hyperthermia agents, and second, to be used as magnetic vehicles for the antitumor drug doxorubicin (DOX). Two different synthetic methodologies (hydrothermal and co-precipitation) were used to obtain MNRs of maghemite and magnetite. They were coated with poly(ethyleneimine) and poly(sodium 4-styrenesulfonate), and loaded with DOX, using the Layer-by-Layer technique. Evidence of the polymer coating and the drug loading was justified by ATR-FTIR and electrophoretic mobility measurements, and the composition of the coated nanorods was obtained by a thermogravimetric analysis. The nanorods were tested as magnetic hyperthermia agents, and it was found that they provided sufficiently large heating rates to be used as adjuvant therapy against solid tumors. DOX loading and release were determined by UV-visible spectroscopy, and it was found that up to 50% of the loaded drug was released in about 5 h, although the rate of release could be regulated by simultaneous application of hyperthermia, which acts as a sort of external release-trigger. Shape control offers another physical property of the particles as candidates to interact with tumor cells, and particles that are not too elongated can easily find their way through the cell membrane.

Assembly of Soft Electrodes and Ion Exchange Membranes for Capacitive Deionization.

The responsible use of water, as well as its reuse and purification, has been a major problem for decades now. In this work, we study a method for adsorbing ions from aqueous solutions on charged interfaces using highly porous electrodes. This water purification process is based on the electric double layer concept, using the method known as capacitive deionization (CDI): If we pump salty solutions through the volume comprised between two porous electrodes while applying a potential difference to them, ions present in the solution are partially removed and trapped on the electrode surfaces. It has been well established that the use of carbon electrodes in combination with ion exchange membranes (membrane-CDI) improves the efficiency of the method above that achieved with bare activated carbon. Another approach that has been tested is based on coating the carbon with polyelectrolyte layers, converting them into "soft electrodes" (SEs). Here we investigate the improvement found when combining SEs with membranes, and it is shown that the amount of ions adsorbed and the ratio between ions removed and electrons transported reach superior values, also associated with a faster kinetics of the process. The method is applied to the partial desalination of up to 100 mM NaCl solutions, something hardly achievable with bare or membrane-covered electrodes. A theoretical model is presented for the ion transport in the presence of both the membrane and the polyelectrolyte coating.

Magnetoliposomes of mixed biomimetic and inorganic magnetic nanoparticles as enhanced hyperthermia agents.

Recently, liposomes have been explored as a potential solution to improve the biocompatibility and the colloidal stability of magnetic nanoparticles. Protocols have been developed for producing magnetoliposomes of magnetite nanoparticles obtained inorganically (MNPs). However, the biomimetic synthesis of magnetite using heterologous proteins from magnetotactic bacteria has become a real alternative to produce novel biomimetic magnetic nanoparticles (BMNPs). Among these, the BMNPs obtained in presence of MamC protein from Magnetococcus marinus MC-1 have been proposed as excellent candidates to be potentially used as drug nanocarriers and as hyperthermia agents. However, their colloidal stability still needs to be improved while maintaining their magnetic properties intact. One possibility explored in this manuscript is to form magnetoliposomes that contain BMNPs. Indeed, the protocols developed for producing magnetoliposomes of MNPs need to be tested and modified to be able to include BMNPs. In this context, a protocol has been developed to produce both magnetoliposomes filled with MNPs and/or BMNPs and their potential as hyperthermia agents was tested. In fact, for the first time, these two types of nanoparticles were mixed in different proportions to test the composition that would optimize such as behaviour as hyperthermia agents. Interestingly, it was observed that the hyperthermia behaviour of the magnetoliposomes greatly improved if they were filled with a mixture of MNPs and BMNPs. These results indicate that these magnetoliposomes display optimal characteristics to become a potential agent for hyperthermia and that the opening of those liposomes could be externally controlled by applying an alternate magnetic field.

Enhancement of Magnetic Hyperthermia by Mixing Synthetic Inorganic and Biomimetic Magnetic Nanoparticles.

In this work we report on the synthesis and characterization of magnetic nanoparticles of two distinct origins, one inorganic (MNPs) and the other biomimetic (BMNPs), the latter based on a process of bacterial synthesis. Each of these two kinds of particles has its own advantages when used separately with biomedical purposes. Thus, BMNPs present an isoelectric point below neutrality (around pH 4.4), while MNPs show a zero-zeta potential at pH 7, and appear to be excellent agents for magnetic hyperthermia. This means that the biomimetic particles are better suited to be loaded with drug molecules positively charged at neutral pH (notably, doxorubicin, for instance) and releasing it at the acidic tumor environment. In turn, MNPs may provide their transport capabilities under a magnetic field. In this study it is proposed to use a mixture of both kinds of particles at two different concentrations, trying to get the best from each of them. We study which mixture performs better from different points of view, like stability and magnetic hyperthermia response, while keeping suitable drug transport capabilities. This composite system is proposed as a close to ideal drug vehicle with added enhanced hyperthermia response.

Electro-Orientation of Silver Nanowires in Alternating Fields.

In this work, we analyze the orientation of silver nanowires immersed in aqueous solutions, under the effect of alternating electric fields in a broad frequency range covering from a few Hz to several MHz. The degree of orientation is experimentally determined by electro-optical techniques, which present the advantage of measuring multiple particles at the same time. In the electro-orientation spectrum, we observe frequency dispersion in the kHz range and provide a theoretical explanation for this behavior: at high frequencies, charge separation in the nanoparticles leads to a large induced dipole responsible for strong orientation. On the other hand, at low frequencies, redistribution of the ions in solution gives rise to an induced double layer that screens the dipolar fields, and as a consequence, the degree of orientation decreases. Moreover, we measure the transient response when the electric field is switched off, from which the size distribution of the polydisperse sample is obtained. The results match those given by electron microscopy determinations.

Formulation and in vitro evaluation of magnetoliposomes as a potential nanotool in colorectal cancer therapy.

Magnetoliposomes (MLPs) offer many new possibilities in cancer therapy and diagnosis, including the transport of antitumor drugs, hyperthermia treatment, detection using imaging techniques, and even cell migration. However, high biocompatibility and functionality after cell internalization are essential to their successful application. We synthesized maghemite nanoparticles (γ-Fe2O3) by oxidizing magnetite cores (Fe3O4) and coating them with phosphatidylcholine (PC) liposomes, obtained using the thin film hydration method, to generate MLPs. The MLPs were tested in vitro, using human tumor and non-tumor colon cell lines, for cytotoxicity, cell uptake and cellular distribution, and magnetically-induced cell mobility. In addition, blood cells biocompatibility studies were performed. The mean size of the MLPs, with a core of γ-Fe2O3 completely surrounded by PC liposomes, was 90 ± 20 nm, showing a soft magnetic character and a great biocompatibility in all the cell lines assayed including blood cells. Prussian blue staining showed a high MLP cell uptake with maximum internalization at 24 h. TEM analysis showed the MLPs surrounded by the cell membrane and in the cell periphery, suggesting internalization by endocytosis and/or macropinocytosis. Interestingly, the mitochondria presented MLP accumulations, particularly in tumor cells. Finally, MLPs within colon cancer cells were able to induce cell migration when a magnetic field was applied in vitro, indicating the functionality of our nanoformulation. A promising biomedical application of these MLPs is anticipated based on their physical, chemical and biological properties.

Determination of the size distribution of non-spherical nanoparticles by electric birefringence-based methods.

The in situ determination of the size distribution of dispersed non-spherical nanoparticles is an essential characterization tool for the investigation and use of colloidal suspensions. In this work, we test a size characterization method based on the measurement of the transient behaviour of the birefringence induced in the dispersions by pulsed electric fields. The specific shape of such relaxations depends on the distribution of the rotational diffusion coefficient of the suspended particles. We analyse the measured transient birefringence with three approaches: the stretched-exponential, Watson-Jennings, and multi-exponential methods. These are applied to six different types of rod-like and planar particles: PTFE rods, goethite needles, single- and double-walled carbon nanotubes, sodium montmorillonite particles and gibbsite platelets. The results are compared to electron microscopy and dynamic light scattering measurements. The methods here considered provide good or excellent results in all cases, proving that the analysis of the transient birefringence is a powerful tool to obtain complete size distributions of non-spherical particles in suspension.

Nano-engineering of biomedical prednisolone liposomes: evaluation of the cytotoxic effect on human colon carcinoma cell lines.

OBJECTIVES: Liposomes have attracted the attention of researchers due to their potential to act as drug delivery systems for cancer treatment. The present investigation aimed to develop liposomes loaded with prednisolone base and the evaluation of the antiproliferative effect on human colon carcinoma cell lines. METHODS: Liposomes were elaborated by following a reproducible thin film hydration technique. The physicochemical characterization of liposomes included photon correlation spectroscopy, microscopy analysis, Fourier transform infrared spectroscopy, rheological behaviour and electrophoresis. On the basis of these data and drug loading values, the best formulation was selected. Stability and drug release properties were also tested. KEY FINDINGS: Resulting liposomes exhibited optimal physicochemical and stability properties, an excellent haemocompatibility and direct antiproliferative effect on human colon carcinoma T-84 cell lines. CONCLUSIONS: This study shows direct antitumour effect of prednisolone liposomal formulation, which opens the door for liposomal glucocorticoids as novel antitumour agents.

Hyperthermia-Triggered Gemcitabine Release from Polymer-Coated Magnetite Nanoparticles.

In this work a combined, multifunctional platform, which was devised for the simultaneous application of magnetic hyperthermia and the delivery of the antitumor drug gemcitabine, is described and tested in vitro. The system consists of magnetite particles embedded in a polymer envelope, designed to make them biocompatible, thanks to the presence of poly (ethylene glycol) in the polymer shell. The commercial particles, after thorough cleaning, are provided with carboxyl terminal groups, so that at physiological pH they present negative surface charge. This was proved by electrophoresis, and makes it possible to electrostatically adsorb gemcitabine hydrochloride, which is the active drug of the resulting nanostructure. Both electrophoresis and infrared spectroscopy are used to confirm the adsorption of the drug. The gemcitabine-loaded particles are tested regarding their ability to release it while heating the surroundings by magnetic hyperthermia, in principle their chances as antitumor agents. The release, with first-order kinetics, is found to be faster when carried out in a thermostated bath at 43 °C than at 37 °C, as expected. But, the main result of this investigation is that while the particles retain their hyperthermia response, with reasonably high heating power, they release the drug faster and with zeroth-order kinetics when they are maintained at 43 °C under the action of the alternating magnetic field used for hyperthermia.