Synthesis of bombesin-functionalized iron oxide nanoparticles and their specific uptake in prostate cancer cells

The imaging of molecular markers associated with disease offers the possibility for earlier detection and improved treatment monitoring. Receptors for gastrin-releasing peptide are overexpressed on prostate cancer cells offering a promising imaging target, and analogs of bombesin, an amphibian tetradecapeptide have been previously demonstrated to target these receptors. Therefore, the pan-bombesin analog [β-Ala11, Phe13, Nle14]bombesin-(7-14) was conjugated through a linker to dye-functionalized superparamagnetic iron oxide nanoparticles for the development of a new potential magnetic resonance imaging probe. The peptide was conjugated via click chemistry, demonstrating a complementary alternative methodology to conventional peptide-nanoparticle conjugation strategies. The peptide-functionalized nanoparticles were then demonstrated to be selectively taken up by PC-3 prostate cancer cells relative to unfunctionalized nanoparticles and this uptake was inhibited by the presence of free peptide, confirming the specificity of the interaction. This study suggests that these nanoparticles have the potential to serve as magnetic resonance imaging probes for the detection of prostate cancer.     

Cellular uptake of folate-conjugated lipophilic superparamagnetic iron oxide nanoparticles

We prepared five folate-conjugated lipophilic superparamagnetic iron oxide nanoparticles (F₅-Liposuperparamagnetic iron oxide nanoparticles(SPIONs), 5.5 and 11 nm) and investigated their cellular uptake with KB cells, which is one of the representative folate-receptor over-expressing human epidermoid carcinoma cells, using MRI. The cellular uptake tests with the respective 5.5 and 11 nm F₅-LipoSPIONs at a fixed particle concentration showed appreciable amount of receptor-mediated uptakes and the specificity was higher in 5.5 nm SPIONs, due to its higher folic acid (FA) density, without inhibition. However, the numbers of the particles taken up under FA inhibition were similar, irrespective of their sizes.     

Synthesis of ultrasmall magnetic iron oxide nanoparticles and study of their colloid and surface chemistry

Colloidal nanoparticles of Fe₃O₄ (4 nm) were synthesized by high-temperature hydrolysis of chelated iron (II) and (III) diethylene glycol alkoxide complexes in a solution of the parent alcohol (H₂DEG) without using capping ligands or surfactants: [Fe(DEG)Cl₂]²⁻+2[Fe(DEG)Cl₃]²⁻+2H₂O+2OH⁻→Fe₃O₄+3H₂DEG+8Cl⁻ The obtained particles were reacted with different small-molecule polydentate ligands, and the resulting adducts were tested for aqueous colloid formation. Both the carboxyl and α-hydroxyl groups of the hydroxyacids are involved in coordination to the nanoparticles’ surface. This coordination provides the major contribution to the stability of the ligand-coated nanoparticles against hydrolysis.     

Synthesis of iron oxide nanoparticles under oxidizing environment and their stabilization in aqueous and non-aqueous media

Synthesis of magnetite (Fe₃O₄) nanoparticles under oxidizing environment by precipitation from aqueous media is not straightforward because Fe²⁺ gets oxidized to Fe³⁺ and thus the ratio of Fe³⁺:Fe²⁺=2:1 is not maintained during the precipitation. A molar ratio of Fe³⁺:Fe²⁺ smaller than 2:1 has been used by many to compensate for the oxidation of Fe²⁺ during the preparation. In this work, we have prepared iron oxide nanoparticles in air environment by the precipitation technique using initial molar ratios Fe³⁺:Fe²⁺?2:1. The phases of the resulting powders have been determined by several techniques. It is found that the particles consist mainly of maghemite with little or no magnetite phase. The particles have been suspended in non-aqueous and aqueous media by coating the particles with a single layer and a bilayer of oleic acid, respectively. The particle sizes, morphology and the magnetic properties of the particles and the ferrofulids prepared from these particles are reported. The average particle sizes obtained from the TEM micrographs are 14, 10 and 9 nm for the water, kerosene and dodecane-based ferrofluids, respectively, indicating a better dispersion in the non-aqueous media. The specific saturation magnetization (σ_s) value of the oleic-acid-coated particles (∼53 emu/g) is found to be lower than that for the uncoated particles (∼63 emu/g). Magnetization σ_s of the dodecane-based ferrofluid is found to be 10.1 emu/g for a volume fraction of particles ?=0.019. Zero coercivity and zero remanance on the magnetization curves indicate that the particles are superparamagnetic (SPM) in nature.     

Preparation of epidermal growth factor (EGF) conjugated iron oxide nanoparticles and their internalization into colon cancer cells

Epidermal growth factor (EGF) was conjugated with carboxymethyldextran (CMDx) coated iron oxide magnetic nanoparticles using carbodiimide chemistry to obtain magnetic nanoparticles that target the epidermal growth factor receptor (EGFR). Epidermal growth factor modified magnetic nanoparticles were colloidally stable when suspended in biological buffers such as PBS and cell culture media. Both targeted and non-targeted nanoparticles were incubated with CaCo-2 cancer cells, known to overexpress EGFR. Nanoparticle localization within the cell was visualized by confocal laser scanning microscopy and light microscopy using Prussian blue stain. Results showed that targeted magnetic nanoparticles were rapidly accumulated in both flask-shaped small vesicles and large circular endocytic structures. Internalization patterns suggest that both clathrin-dependent and clathrin-independent receptors mediated endocytosis mechanisms are responsible for nanoparticle internalization.     

Synthesis and characterization of superparamagnetic iron oxide nanoparticles for biomedical applications

Monodisperse iron oxide nanoparticles (NPs) of 4 nm were obtained through high-temperature solution phase reaction of iron (III) acetylacetonate with 1, 2-hexadecanediol in the presence of oleic acid and oleylamine. The as-synthesized iron oxide nanoparticles have been characterized by X-ray diffraction, transmission electron microscopy, Mössbauer spectroscopy and magnetic measurements. The species obtained were Fe₃O₄ and/or $\upgamma$ -Fe₂O₃. These NPs are superparamagnetic at room temperature and even though the reduced particle size they show a high saturation magnetization (MS ≈ 90 emu/g).     

Recovery of iron/iron oxide nanoparticles from solution: comparison of methods and their effects

Most methods currently being used to recover Fe⁰-core/oxide-shell nanoparticles from solutions (including the solvents they are synthesized or stored in) are potentially problematic because they may alter the particle composition (e.g., depositing salts formed from solutes) or leave the particles prone to transformations during subsequent storage and handling (e.g., due to residual moisture). In this study, several methods for recovery of nanoparticles from aqueous solution were studied to determine how they affect the structure and reactivity of the recovered materials. Simple washing of the nanoparticles during vacuum filtration (i.e., “flash drying”) can leave up to ~17 wt% residual moisture. Modeling calculations suggest this moisture is mostly capillary or matric water held between particles and particle aggregates, which can be removed by drying for short periods at relative vapor pressures below 0.9. Flash drying followed by vacuum drying, all under N₂, leaves no detectable residue from precipitation of solutes (detectable by X-ray photoelectron spectroscopy, XPS), no significant changes in overall particle composition or structure (determined by transmission electron microscopy, TEM), and negligible residual moisture (by thermogravimetric analysis, TGA). While this improved flash-drying protocol may be the preferred method for recovering nanoparticles for many purposes, we found that Fe⁰-core/oxide-shell nanoparticles still exhibit gradual aging during storage when characterized electrochemically with voltammetry.     

Synthesis of α- and β-FeOOH iron oxide nanoparticles in non-ionic surfactant medium

Forced hydrolysis of Fe(III) ions in acidic media was performed under controlled synthetic conditions to produce α- and β-FeOOH iron oxides. The forced hydrolysis synthesis was carried out, separately, in an aqueous medium and the lamellar lyotropic liquid crystalline phase of a commercial non-ionic surfactant/water system. The FT-IR analyses confirmed formation of α- and β-FeOOH iron oxides in the aqueous and the surfactant media with slight formation of ferrihydrite and haematite. TEM micrographs have shown that particles formed in the lamellar lyotropic phase are smaller than those produced in the aqueous medium with their smallest size dimension being constrained in the nanometre scale with a size ranging between 5 and 100 nm. Particles produced in the nanoscale size appeared to have different optical properties compared to their counterparts produced in the microscale size.     

Synthesis and characterization of hybrid nanocomposites of polypyrrole filled with iron oxide nanoparticles

Hybrid polypyrrole (PPy)/α-Fe₂O₃ nanocomposite films were fabricated by spin coating on a glass substrate. X-Ray diffraction analysis revealed the crystalline structure of α-Fe₂O₃ nanostructures and the nanocomposites. The broad PPy peak weakened in intensity as the α-Fe₂O₃ content increased in PPy/α-Fe₂O₃ nanocomposites. Characteristic Fourier-transform IR peaks for pure PPy shifted to higher wavenumbers on addition of α-Fe₂O₃ to PPy/α-Fe₂O₃ nanocomposites. This can be attributed to better conjugation and interactions between PPy and α-Fe₂O₃ nanoparticles. Field-emission scanning electron microscopy, transmission electron microscopy, and atomic force microscopy images of the nanocomposites reveal a uniform distribution of α-Fe₂O₃ nanoparticles in the PPy matrix. UV-vis absorption spectroscopy revealed a blue shift from λ_max= 441 nm for PPy to λ_max= 392 nm for PPy/α-Fe₂O₃, reflecting strong interactions between PPy and α-Fe₂O₃ nanoparticles. The room-temperature dc electrical conductivity increased from 4.33×10⁻⁹ to 1.81×10⁻⁸ S/cm as the α-Fe₂O₃ nanoparticle content increased from 10 to 50 wt.% in PPy/α-Fe₂O₃ nanocomposites.     

Tannin biosynthesis of iron oxide nanoparticles

In this work, iron oxide nanoparticles synthesized with gallic acid and tannic acid are characterized using High-Resolution Transmission Electron Microscopy (HRTEM). Its size, form, and structure are compared with nanoparticles obtained previously using alfalfa biomass in order to find a simpler, consistent, and environmentally friendly method in the production of iron oxide nanoparticles.     

Characterization of magnetic iron oxide nanoparticles

Magnetic nanoparticles have promising applications in many areas, for example optics, electronics, biology, medicine, etc. The main goal of this study is to synthezise and characterize ε-Fe₂O₃ magnetic nanoparticles embedded in amorphous SiO₂ matrix. The Mössbauer spectroscopy analysis of the samples was complemented by the study of X-ray powder diffraction and high-resolution transmission electron microscopy.     

Nanosized cancer cell-targeted polymeric immunomicelles loaded with superparamagnetic iron oxide nanoparticles

Stable 30–50 nm polymeric polyethylene glycol–phosphatidylethanolamine (PEG–PE)-based micelles entrapping superparamagnetic iron oxide nanoparticles (SPION) have been prepared. At similar concentrations of SPION, the SPION-micelles had significantly better magnetic resonance imaging (MRI) T2 relaxation signal compared to ‘plain’ SPION. Freeze-fracture electron microscopy confirmed SPION entrapment in the lipid core of the PEG–PE micelles. To enhance the targeting capability of these micelles, their surface was modified with the cancer cell-specific anti-nucleosome monoclonal antibody 2C5 (mAb 2C5). Such mAb 2C5-SPION immunomicelles demonstrated specific binding with cancer cells in vitro and were able to bring more SPION to the cancer cells thus demonstrating the potential to be used as targeted MRI contrast agents for tumor imaging.     

Magnetic iron oxide nanoparticles: Synthesis and surface coating techniques for biomedical applications

Iron oxide nanoparticles are the most popular magnetic nanoparticles used in biomedical applications due to their low cost, low toxicity, and unique magnetic property. Magnetic iron oxide nanoparticles, including magnetite （Fe304） and maghemite （γ-Fe203）, usually exhibit a superparamagnetic property as their size goes smaller than 20 nm, which are often denoted as superparamagnetic iron oxide nanoparticles （SPIONs） and utilized for drug delivery, diagnosis, therapy, and etc. This review article gives a brief introduction on magnetic iron oxide nanoparticles in terms of their fundamentals of magnetism, magnetic resonance imaging （MRI）, and drug delivery, as well as the synthesis approaches, surface coating, and application examples from recent key literatures. Because the quality and surface chemistry play important roles in biomedical applications, our review focuses on the synthesis approaches and surface modifications of iron oxide nanopar- ticles. We aim to provide a detailed introduction to readers who are new to this field, helping them to choose suitable synthesis methods and to optimize the surface chemistry of iron oxide nanoparticles for their interests.     

Methodology description for detection of cellular uptake of PVA coated superparamagnetic iron oxide nanoparticles (SPION) in synovial cells of sheep

The detection of superparamagnetic iron oxide nanoparticles (SPION) in synoviocytes is reported. Synoviocytes were incubated for 2, 12, 24 and 48 h with 1.5 mg/ml of PVA coated SPION under the influence of magnets (12 h). Particles were well tolerated by the synoviocytes, were easily detected using the Turnbulls and Prussian blue reactions between 12 and 24 h.     

Effect of surface charge on the colloidal stability and in vitro uptake of carboxymethyl dextran-coated iron oxide nanoparticles

An easy and novel routine are reported for the preparation of metallic silver nanoparticles (AgNPs) with controlled morphology, using Na⁺–magadiite swelled with hexadecyltrimethylammonium bromide (CTA⁺–magadiite) and a layered aluminophosphate with kanemite-type structure modified with n-dodecylammonium and n-butylammonium (but,dod-AlPO-kan) as hosts. For the preparation of the metallic AgNPs (Ag⁰) in the interlamellar space, the CTA⁺–magadiite and but,dod-AlPO-kan hosts were dispersed in N,N-dimethylformamide (DMF) solution with different AgNO₃ concentrations. DMF acts as reducing agent of Ag⁺ ions leading to nanoparticles with disk-like morphology of magadiite silicate; these were characterized by TEM and UV–Vis spectroscopy. On the other hand, the AgNPs are intercalated in but,dod-AlPO-kan showing spherical-like morphology. The UV–Vis spectra of the nanocomposites based on Ag⁰ and magadiite silicate show bands at 565 nm that can be attributed to Ag⁰ nanodisks. The Ag-but,dod-AlPO-kan-based nanocomposites present a band at 422 nm attributed to the surface plasmon resonance of Ag⁰ nanospheres. The results of transmission electron microscopy agree very well with XRD and UV–Vis analysis, indicating the formation of AgNPs with different morphologies using the two kinds of lamellar materials. The magadiite host has an important role in the synthesis of Ag nanodisks, because it controls the growth of nanoparticles inside the interlayer region with disk-like morphology due the high interlayer interactions of the silicate, leading to the growth of nanoparticles in only two directions (xy plane). On the other hand, when but,dod-AlPO-kan is used a sphere-like morphology is preferred due the best accommodation of AgNPs between the layers of aluminophosphate host.     

Synthesis of amino-group functionalized superparamagnetic iron oxide nanoparticles and applications as biomedical labeling probes

Superparamagnetic iron oxide (SPIO) nanoparticles were synthesized by coprecipitation technique and further functionalized with amino-group to obtain amino-group functionalized (amino-SPIO) nanoparticles. The X-ray diffraction results reveal the structure of amino-SPIO nanoparticles, from which the average iron core diameter is approximately 10 nm by calculation; while Zetasizer reveals their hydrodynamic diameter are mainly distributed in the range of 40?C60 nm. These nanoparticles can be taken up by liver tissue, resulting in dramatically darkening of liver tissue under T2-magnetic resonance imaging (MRI). The spin?Cspin relaxivity coefficient of these nanoparticles is 179.20 mM^?1 s^?1 in a 1.5 T magnetic resonance system. In addition, amino-SPIO nanoparticles were conjugated to Tat (FITC) peptide and incubated with neural stem cells in vitro, the authors can detect the positive-labeling (labeled) neural stem cells showing green fluorescence, which indicates Tat (FITC) peptide-derivated amino-SPIO nanoparticles are able to enter cells. Furthermore, it was also find significant negative T2 contrast enhancement when compared with the non-nanoparticles-labeled neural stem cells in T2-weighted MRI. The amino-SPIO nanoparticles show promising potential as a new type of labeling probes, which can be used in magnetic resonance-enhanced imaging and fluorescence diagnosis.     

Size-dependent magnetic properties of iron oxide nanoparticles

Uniform iron oxide nanoparticles in the size range from 10 to 24 nm and polydisperse 14 nm iron oxide particles were prepared by thermal decomposition of Fe(III) carboxylates in the presence of oleic acid and co-precipitation of Fe(II) and Fe(III) chlorides by ammonium hydroxide followed by oxidation, respectively. While the first method produced hydrophobic oleic acid coated particles, the second one formed hydrophilic, but uncoated, nanoparticles. To make the iron oxide particles water dispersible and colloidally stable, their surface was modified with poly(ethylene glycol) and sucrose, respectively. Size and size distribution of the nanoparticles was determined by transmission electron microscopy, dynamic light scattering and X-ray diffraction. Surface of the PEG-functionalized and sucrose-modified iron oxide particles was characterized by Fourier transform infrared (FT-IR) and Raman spectroscopy and thermogravimetric analysis (TGA). Magnetic properties were measured by means of vibration sample magnetometry and specific absorption rate in alternating magnetic fields was determined calorimetrically. It was found, that larger ferrimagnetic particles showed higher heating performance than smaller superparamagnetic ones. In the transition range between superparamagnetism and ferrimagnetism, samples with a broader size distribution provided higher heating power than narrow size distributed particles of comparable mean size. Here presented particles showed promising properties for a possible application in magnetic hyperthermia.     

Doxorubicin delivered to MCF-7 cancer cells by superparamagnetic iron oxide nanoparticles: effects on subcellular distribution and cytotoxicity

The clinical use of the anticancer drug doxorubicin (DOX) is limited by strong side effects and phenomena of cell resistance. Drug targeting by binding DOX to nanoparticles could overcome these limitations. We recently described a method to associate DOX to superparamagnetic iron oxide nanoparticles (SPION) in view of magnetic drug targeting (Munnier et al. in Int J Pharm 363:170–176, 2008). DOX is bound to the nanoparticle surface through a pre-formed DOX–Fe²⁺ complex. The DOX-loaded SPION present interesting properties in terms of drug loading and biological activity in vitro. The purpose of this study is to explore the possible mechanisms of the in vitro cytotoxicity of DOX-loaded SPION. The uptake of SPION was followed qualitatively by conventional optical microscopy after Prussian blue staining and quantitatively by iron determination by atomic absorption spectroscopy. The subcellular distribution of intrinsically fluorescent DOX was followed by confocal spectral imaging (CSI) and the subsequent cytotoxicity by the MTT method. We reveal modifications of DOX intracellular interactions for SPION-delivered drug and increased cytotoxicity. These results are discussed in terms of internalization route of the drug and of a potential role of iron oxide nanoparticles in the observed cytotoxicity.     

Microdistribution of copper-carbonate and iron oxide nanoparticles in treated wood

Aqueous dispersions of copper-carbonate nanoparticles and microparticles have just begun to be exploited commercially for the preservative treatment of wood. The success of the new systems will depend, in part, on the uniform distribution of the preservative in wood and the ability of copper to penetrate cell walls. We examined the distribution of copper in wood treated with a nano-Cu preservative. Copper particles are not uniformly distributed in treated wood, but they accumulate in voids that act as the flow paths for liquids in wood. Particles are deposited on, but not within cell walls. Nevertheless, elemental copper is present within cell walls, but at a lower level than that in wood treated with a conventional wood preservative. These findings suggest that nano-Cu preservatives are able to deliver bioactive components into wood cell walls even though the majority of copper particles are too large to penetrate the cell wall’s nanocapillary network.     

Nitrilimine cycloadditions catalyzed by iron oxide nanoparticles

Nitrilimine cycloadditions to ethylenes, acetylenes, and activated nitriles have been exploited in the presence of catalytic amounts of oleic-acid-coated iron oxide nanoparticles (diameter?=?11.9?±?1.0 nm). The reactions were fully regioselective with monosubstituted ethylenes and ethyl cyanoformiate, while mixtures of cycloadducts were obtained in the presence of methyl propiolate. The intervention of iron oxide nanoparticles allowed carrying out the cycloadditions at milder conditions compared to the metal-free thermal processes. A labile intermediate has been proposed to explain this behavior.

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Graphical abstract Nitrilimine cycloadditions to ethylenes, acetylenes, and activated nitriles have been exploited in the presence of catalytic amounts of oleic-acid-coated iron oxide nanoparticles.