J.L. Viota

Nanoengineering of doxorubicin delivery systems with functionalized maghemite nanoparticles

Superparamagnetic iron oxide nanoparticles are developing as promising candidates for biomedical applications such as targeted drug delivery. In particular, they represent an alternative to existing antitumor drug carriers, because of their ultra-fine size, low toxicity and magnetic characteristics. Nevertheless, there is a need to functionalize them in order to achieve good biocompatibility, efficient modification for further attachment of biomolecules, and improved stability. In this work we describe the functionalization of superparamagnetic maghemite nanoparticles encapsulated in a silica shell. After their chemical modification with positive (3-aminopropyl)trimethoxysilane, a gold layer was deposited in order to facilitate incorporation of the antitumor drug, doxorubicin (DOX), up to a maximum loading of 80 μmol/g. In vitro cell uptake of nanocomposites was performed with DLD-1 colon cancer cells and PLC-PRF-5 liver cancer cells. Confocal microscopy photos illustrate that doxorubicin-loaded nanoparticles accumulate in both the cytoplasm and the cell nuclei. Cell survival efficiency with maghemite nanocomposites was determined via the MTT assay, and the cytotoxicity study proved that they exhibited significant toxicity against both types of cancer cells, although the improvement over free DOX treatment is more evident in the case of DLD-1 cancer cells when the most dilute drug and particle solutions are compared.

Study of the magnetorheological response of aqueous magnetite suspensions stabilized by acrylic acid polymers

In this paper we describe the magnetorheological (MR) behavior of aqueous suspensions consisting of magnetite particles stabilized by poly(acrylic acid) polymers (PAA). A previous work on the colloidal stability of the same systems for different pH values and polymer concentrations demonstrated that the addition of PAA polymers has a very significant effect on the stability. In the present contribution, we study the MR effect of the suspensions stabilized by two different commercial polymers, as a function of pH, magnetic field strength and magnetite volume fraction. All the results are discussed in terms of the interfacial properties of the systems. It is demonstrated that for a given concentration of micrometer particles, the rheological response strongly depends on pH, on the volume fraction of magnetite particles, on the type of polymer added for increasing the stability and on the magnetic field strength. Changing the polymer used provokes clear rheological differences for the same sample conditions (field strength, volume fraction and pH). This is suggested to be due to the hydrophobic/hydrophilic balance of the polymer affecting the magnetic field ability to form magnetic structures by aggregation of the magnetized particles. The results are compared to the predictions of the so-called standard chain model, based on the assumption that the MR effect is the result of the balance between the magnetic interactions (tending to establish some degree of order in the suspension by formation of particle chains in the direction of the field) and hydrodynamic ones (tending to destroy the formed structures by viscous stress on the chains). It is found that the behavior of the yield stress does not agree well with the predictions of the model when the relative proportion of both particle and polymer confers optimum stability to the dispersions. This is likely due to the fact that the presence of the stabilizing polyelectrolyte provokes that the magnetic field is not as effective in structuring the suspension as deduced from the chain model.

Functionalized magnetic nanoparticles as vehicles for the delivery of the antitumor drug gemcitabine to tumor cells. Physicochemical in vitro evaluation

Gemcitabine is a chemotherapy drug used in different carcinomas, although because it displays a short biological half-life, its plasmatic levels can quickly drop below the effective threshold. Nanoparticle-based drug delivery systems can provide an alternative approach for regulating the bioavailability of this and most other anticancer drugs. In this work we describe a new model of composite nanoparticles consisting of a core of magnetite nanoparticles, coated with successive layers of high molecular weight poly(acrylic acid) and chitosan, and a final layer of folic acid. The possibility of using these self-assembled nanostructures for gemcitabine vehiculization is explored. First, the surface charge of the composite particles is studied by means of electrophoretic mobility measurements as a function of pH for poly(acrylic acid) (carbopol) of different molecular weights. The adsorption of folic acid, aimed at increasing the chances of the particles to pass the cell membrane, is followed up by optical absorbance measurements, which were also employed for drug adsorption determinations. As a main result, it is shown that gemcitabine adsorbs onto the surface of chitosan/carbopol-coated magnetite nanoparticles. In vitro experiments show that the functionalized magnetic nanoparticles are able to deliver the drug to the nuclei of liver, colon and breast tumor cells.

Preparation of multi-functionalized Fe3O4/Au nanoparticles for medical purposes

In this work, we investigate a route towards the synthesis of multi-functionalized nanoparticles for medical purposes. The aim is to produce magnetite/gold (Fe3O4/Au) nanoparticles combining several complementary properties, specifically, being able to carry simultaneously an antitumor drug and a selected antibody chosen so as to improve specificity of the drug vehicle. The procedure included, firstly, the preparation of Fe3O4 cores coated with Au nanoparticles: this was achieved by using initially the layer-by-layer technique in order to coat the magnetite particles with a three polyelectrolyte (cationic-anionic-cationic) layer. With this, the particles became a good substrate for the growth of the gold layer in a well-defined core-shell structure. The resulting nanoparticles benefit from the magnetic properties of the magnetite and the robust chemistry and the biostability of gold surfaces. Subsequently, the Fe3O4/Au nanoparticles were functionalized with a humanized monoclonal antibody, bevacizumab, and a chemotherapy drug, doxorubicin. Taken together, bevacizumab enhances the therapeutic effect of chemotherapy agents on some kinds of tumors. In this work we first discuss the morphology of the particles and the electrical characteristics of their surface in the successive synthesis stages. Special attention is paid to the chemical stability of the final coating, and the physical stability of the suspensions of the nanoparticles in aqueous solutions and phosphate buffer. We describe how optical absorbance and electrokinetic data provide a follow up of the progress of the nanostructure formation. Additionally, the same techniques are employed to demonstrate that the composite nanoparticles are capable of loading/releasing doxorubicin and/or bevacizumab.

Electrophoretic characterization of insulin growth factor (IGF-1) functionalized magnetic nanoparticles.

The synthesis of composite nanoparticles consisting of a magnetite core coated with a layer of the hormone insulin growth factor 1 (IGF-1) is described. The adsorption of the hormone in the different formulations is first studied by electrophoretic mobility measurements as a function of pH, ionic strength, and time. Because of the permeable character expected for both citrate and IGF-1 coatings surrounding the magnetite cores, an appropriate analysis of their electrophoretic mobility must be addressed. Recent developments of electrokinetic theories for particles covered by soft surface layers have rendered possible the evaluation of the softness degree from raw electrophoretic mobility data. In the present contribution, the data are quantitatively analyzed based on the theoretical model of the electrokinetics of soft particles. As a result, information is obtained on both the thickness and the charge density of the surrounding layer. It is shown that IGF-1 adsorbs onto the surface of citrate-coated magnetite nanoparticles, and adsorption is confirmed by dot-blot analysis. In addition, it is also demonstrated that the external layer of IGF-1 exerts a shielding effect on the surface charge of citrate-magnetite particles, as suggested by the mobility reduction upon contacting the particles with the hormone. Aging effects are demonstrated, providing an electrokinetic fingerprint of changes in adsorbed protein configuration with time.

Synergistic cytotoxicity of the poly (ADP-ribose) polymerase inhibitor ABT-888 and temozolomide in dual-drug targeted magnetic nanoparticles.

Hepatocellular carcinoma (HCC) is associated with a poor prognosis because of a lack of effective treatment options. The objective of this study was to examine a new strategy for HCC treatment, namely the use of poly (ADP‐ribose) polymerase 1 (PARP‐1) inhibitor (ABT‐888) together with Temozolomide (TMZ) incorporated onto magnetic nanoparticles.