Citrate-coated Iron Oxide Nanoparticles Research Articles

In vitro and in vivo acute toxicity of a novel citrate-coated magnetite nanoparticle.

Magnetic nanoparticles (MNps) have become powerful tools for multiple biomedical applications such as hyperthermia drivers, magnetic resonance imaging (MRI) vectors, as well as drug-delivery systems. However, their toxic effects on human health have not yet been fully elucidated, especially in view of their great diversity of surface modifications and functionalizations. Citrate-coating of MNps often results in increased hydrophilicity, which may positively impact their performance as drug-delivery systems. Nonetheless, the consequences on the intrinsic toxicity of such MNps are unpredictable. Herein, novel magnetite (Fe3O4) nanoparticles covered with citrate were synthesized and their potential intrinsic acute toxic effects were investigated using in vitro and in vivo models. The proposed synthetic pathway turned out to be simple, quick, inexpensive, and reproducible. Concerning toxicity risk assessment, these citrate-coated iron oxide nanoparticles (IONps) did not affect the in vitro viability of different cell lines (HaCaT and HepG2). Moreover, the in vivo acute dose assay (OECD test guideline #425) showed no alterations in clinical parameters, relevant biochemical variables, or morphological aspects of vital organs (such as brain, liver, lung and kidney). Iron concentrations were slightly increased in the liver, as shown by Graphite Furnace Atomic Absorption Spectrometry and Perls Prussian Blue Staining assays, but this finding was considered non-adverse, given the absence of accompanying functional/clinical repercussions. In conclusion, this study reports on the development of a simple, fast and reproducible method to obtain citrate-coated IONps with promising safety features, which may be used as a drug nanodelivery system in the short run. (263 words).

Recovery trend to co-exposure of iron oxide nanoparticles (γ-Fe2O3) and glyphosate in liver tissue of the fish Poecilia reticulata

Citrate-coated iron oxide nanoparticles (IONPs) have potential use in environmental remediation, with possibilities in decontaminating aquatic environments exposed to toxic substances. This study analyzed IONPs associated to Roundup Original, a glyphosate-based herbicide (GBH), and pure glyphosate (GLY), through ultrastructural and histopathological biomarkers in liver tissue, from females of Poecilia reticulata exposed to: iron ions (0.3 mg/L) (IFe) and IONPs (0.3 mgFe/L) associated with GLY (0.65 mg/L) and GBH (0.65 mgGLY/L (IONP + GBH1) and 1.30 mgGLY/L (IONP + GBH2)) for a period of 7, 14 and 21 days, followed by an equal post-exposure period only in reconstituted water. For the assays, the synthetized IONPs had crystalline and rounded shape with an average diameter of 2,90 nm, hydrodynamic diameter 66,6 mV, zeta potential −55,4 and diffraction profile of maghemite (γ-Fe2O3). The data obtained by biomarkers indicated a high inflammatory response in all treatments. These same parameters, considered during the post-exposure period indicated recovery in reaction patterns of circulatory disturbances and regressive changes, resulting in average reductions of 37,53 points in IFe, 21 points in IONP + GBH1, 15 points in IONP + GBH2 and 11 points in IONP + GLY in total histopathological index of liver after 21 days post-exposure. However, although the cellular and tissue responses were significant, there was no change in the condition factor and hepatosomatic index, denoting resilience of the experimental model.

Initial interaction of citrate-coated iron oxide nanoparticles with the glycocalyx of THP-1 monocytes assessed by real-time magnetic particle spectroscopy and electron microscopy

Interaction with biological material can alter physicochemical parameters of magnetic nanoparticles and might thereby change their magnetic behavior with potentially important implications for various nanoparticle applications. Little is known about changes of the magnetic behavior that occur during the initial phase of cell binding and uptake. We investigate the magnetic behavior of very small superparamagnetic iron-oxide nanoparticles (VSOP) during initial contact with THP-1 monocytes. We combine real-time magnetic particle spectroscopy (MPS), a fast and sensitive method for specific detection of magnetic nanoparticles in biological specimen with high-pressure-freezing/freeze-substitution transmission electron microscopy (HPF/FS-TEM), enabling us to generate snapshots of the interaction of VSOP with the cellular glycocalyx. MPS reveals significant changes of the dynamic magnetic behavior within seconds after VSOP injection into monocyte suspensions that correlate with the formation of nanoparticle clusters in the glycocalyx. The combination of real-time MPS and HPF/FS-TEM provides an ideal platform to analyze magnetic behaviors of nanoparticles upon interaction with cells and tissues.

Erratum to: Uptake of citrate-coated iron oxide nanoparticles into atherosclerotic lesions in mice occurs via accelerated transcytosis through plaque endothelial cells

Erratum to: Uptake of citrate-coated iron oxide nanoparticles into atherosclerotic lesions in mice occurs via accelerated transcytosis through plaque endothelial cells

Uptake of citrate-coated iron oxide nanoparticles into atherosclerotic lesions in mice occurs via accelerated transcytosis through plaque endothelial cells

Abstract Very small superparamagnetic iron oxide nanoparticles (VSOPs) rapidly accumulate in atherosclerotic lesions, thereby enabling plaque visualization by magnetic resonance imaging (MRI). This study was performed to identify the uptake mechanisms of VSOPs into atherosclerotic plaques. Low-density lipoprotein receptor-deficient (LDLR−/−) mice with advanced atherosclerosis were analyzed using MRI and transmission electron microscopy (TEM) at various time points after intravenous administration of VSOPs. Post-mortem MRI detected VSOP labeling of atherosclerotic plaques 10 min after injection, and the signal increased over the first 3 h. TEM revealed that the intensive plaque labeling was mediated by accelerated transcytosis of VSOPs through endothelial cells overlaying atherosclerotic lesions. Experiments with endocytosis inhibitors and small interfering RNA (siRNA) revealed a dynamin-dependent mechanism involving both clathrin- and caveolin-mediated processes. In cell culture experiments, endothelial VSOP uptake was enhanced under proatherogenic flow and TNFα stimulation, conditions that are both present in plaque areas. Our study demonstrates that VSOPs enable non-invasive MRI assessment of accelerated endothelial transcytosis, an important pathomechanism in atherosclerotic plaque formation.

Anisotropic magnetic field observed at 300 K in citrate-coated iron oxide nanoparticles: effect of counterions

Iron oxide nanoparticles (IONPs) have been synthesized by chemical co-precipitation method and coated with three citrates, namely, tri-lithium citrate (TLC), tri-sodium citrate (TSC), or tri-potassium citrate (TKC). In these ‘core–shell’ structures, the ‘core’ is a cluster of average 3 IONPs which is enveloped by a ‘shell’ of citrate molecules and counterions, and thus called ‘core–shell’ nano-clusters (CS-NCs), of average size ~20 to 22 nm. The counterions in the three CS-NCs differ in ionic radii (\(r_{\text{ion}}\)), in the order of Li+ 300 K for TLC-coated CS-NCs and <300 K for TSC- and TKC-coated CS-NCs. The EMR linewidth (∆B PP), measured at 300 K, was also the broadest for TLC-coated CS-NCs. At low temperatures, ΔB PP was found to increase more significantly for TSC- and TKC-coated CS-NCs than for TLC-coated CS-NCs. These results indicate a significant anisotropic field effect; arising due to thermal motion of counterions at 300 K, on the magnetic interactions in TLC-coated CS-NCs. To our knowledge, this is the first report on the effect of counterions on magnetic interactions between CS-NCs.

RETRACTED: Rapid synthesis of water-dispersible superparamagnetic iron oxide nanoparticles by a microwave-assisted route for safe labeling of endothelial progenitor cells

We synthesize highly crystalline citrate-coated iron oxide superparamagnetic nanoparticles that are stable and readily dispersible in water by an extremely fast microwave-assisted route and investigate the uptake of magnetic nanoparticles by endothelial cells. Nanoparticles form large aggregates when added to complete endothelial cell medium. The size of the aggregates was controlled by adjusting the ionic strength of the medium. The internalization of nanoparticles into endothelial cells was then investigated by transmission electron microscopy, magnetometry and chemical analysis, together with cell viability assays. Interestingly, a sevenfold more efficient uptake was found for systems with larger nanoparticle aggregates, which also showed significantly higher magnetic resonance imaging effectiveness without compromising cell viability and functionality. We are thus presenting an example of a straightforward microwave synthesis of citrate-coated iron oxide nanoparticles for safe endothelial progenitor cell labeling and good magnetic resonance cell imaging with potential application for magnetic cell guidance and in vivo cell tracking.

Citrate coated iron oxide nanoparticles with enhanced relaxivity for in vivo magnetic resonance imaging of liver fibrosis

Superparamagnetic iron oxide nanoparticles are widely used for the magnetic resonance imaging (MRI) applications. The surface characteristics, magnetic properties, size and targeting efficiency of the material are crucial factors for using the same as contrast agents. We report a simple synthesis method of citrate coated iron oxide nanoparticles and its systematic characterization. The developed system is highly water dispersible with an average particle size of 12nm. The particles in water are monodisperse and are found to be stable over long periods. The efficiency of the material to de-phase water proton has been studied for various concentrations of iron using longitudinal (T1) and transverse (T2) weighted MRI. The coating thickness of the nanoparticle was optimized so that they exhibited a high transverse to longitudinal relaxivity (r2/r1) ratio of 37.92. A clear dose-dependent contrast enhancement was observed in T2 weighted in vivo MR imaging of liver fibrosis model in rodents. The labelling efficacy of the particle and the intracellular magnetic relaxivity were also investigated and presented. The particles were also tested for blood and cellular compatibility studies. Development of fibrosis and presence of iron in the liver was confirmed by histopathological analysis. From this study, we conclude that the citrate coated ultra small superparamagnetic iron oxide nanoparticles (C-USPION) with optimized parameters like particle size and magnetic property are capable of producing good MR contrast in imaging of liver diseases.

Synthesis and Morphology Control of Gold/Iron Oxide Magnetic Nanocomposites via a Simple Aqueous Method

Gold/iron oxide nanocomposites offer the promise of combining the unique traits of both gold and iron oxide nanomaterials to enable new applications. In this work, a simple and safe method has been established whereby the morphology of gold material in gold/iron oxide nanocomposites can be controlled to be nanowire or nanosphere by adjusting the initial molar ratio of citrate-coated iron oxide nanoparticles to auric acid. Gold/iron oxide nanowires and nanospheres were synthesized through this method. Their structural, optical, and magnetic properties were characterized by transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), ultraviolet-visible spectroscopy (UV-Vis), and vibrating sample magnetometry (VSM). These materials substantially maintained the optical properties of gold nanomaterials and magnetic features of iron oxide magnetic nanoparticles.

Can Magnetic Targeting of Magnetically Labeled Circulating Cells Optimize Intramyocardial Cell Retention?

Therapeutic intracavitary stem cell infusion currently suffers from poor myocardial homing. We examined whether cardiac cell retention could be enhanced by magnetic targeting of endothelial progenitor cells (EPCs) loaded with iron oxide nanoparticles. EPCs were magnetically labeled with citrate-coated iron oxide nanoparticles. Cell proliferation, migration, and CXCR4 chemokine receptor expression were assessed in different labeling conditions and no adverse effects of the magnetic label were observed. The magnetophoretic mobility of labeled EPCs was determined in vitro, with the same magnet as that subsequently used in vivo. Coronary artery occlusion was induced for 30 min in 36 rats (31 survivors), followed by 20 min of reperfusion. The rats were randomized to receive, during brief aortic cross-clamping, direct intraventricular injection of culture medium (n = 7) or magnetically labeled EPCs (n = 24), with (n = 14) or without (n = 10) subcutaneous insertion of a magnet over the chest cavity (n = 14). The hearts were explanted 24 h later and engrafted cells were visualized by magnetic resonance imaging (MRI) of the heart at 1.5 T. Their abundance in the myocardium was also analyzed semiquantitatively by immunofluorescence, and quantitatively by real-time polymerase chain reaction (RT-PCR).Although differences in cell retention between groups failed to be statistically significant using RT-PCR quantification, due to the variability of the animal model, immunostaining showed that the average number of engrafted EPCs was significantly ten times higher with than without magnetic targeting. There was thus a consistent trend favoring the magnet-treated hearts, thereby suggesting magnetic targeting as a potentially new mean of enhancing myocardial homing of intravascularly delivered stem cells. Magnetic targeting has the potential to enhance myocardial retention of intravascularly delivered endothelial progenitor cells.

High-Resolution 1.5-Tesla Magnetic Resonance Imaging for Tissue-Engineered Constructs: A Noninvasive Tool to Assess Three-Dimensional Scaffold Architecture and Cell Seeding

Tissue-engineered scaffolds are made of biocompatible polymers with various structures, allowing cell seeding, growth, and differentiation. Noninvasive imaging methods are needed to study tissue-engineered constructs before and after implantation. Here, we show that high-resolution magnetic resonance imaging (MRI) performed on a clinical 1.5-T device is a reliable technique to assess three-dimensional structures of porous scaffolds and to validate cell-seeding procedures. A high-temperature superconducting detection coil was used to achieve a resolution of 30 x 30 x 30 microm(3) when imaging the scaffolds. Three types of structures with tuneable architectures were prepared from naturally derived polysaccharides and evaluated as scaffolds for mesenchymal stem cell (MSC) culture. To monitor cell seeding, MSCs were magnetically labeled using simple incubation with anionic citrate-coated iron-oxide nanoparticles for 30 min. Iron uptake was quantified using single-cell magnetophoresis, and cell proliferation was checked for 7 days after labeling. Three-dimensional (3D) microstructures of scaffolds were assessed using MRI, revealing lamellar or globular porous organization according to the scaffold preparation process. MSCs with different iron load (5, 12 and 31 pg of iron per cell) were seeded on scaffolds at low density (132 cells/mm(3)) and detected on 3D gradient-echo MR images according to phase distortions and areas of intensely low signal, whose size increased with cell iron load and echo time. Overall signal loss in the scaffold correlated with the number of seeded cells and their iron load. Different organizations of cells were observed depending on the scaffold architecture. After subcutaneous implantation in mice, scaffolds seeded with labeled cells could be distinguished in vivo from scaffold with nonlabeled cells by observation of signal and phase heterogeneities and by measuring the global signal loss. High-resolution 1.5-T MRI combined with efficient intracellular contrast agents shows promise for noninvasive 3D visualization of tissue-engineered constructs before and after in vivo implantation.

Accumulation of Iron Oxide Nanoparticles by Cultured Brain Astrocytes

Magnetic iron oxide nanoparticles are considered for various diagnostic and therapeutic applications in brain including their use as contrast agent for magnetic resonance imaging or as tool for magnetic drug delivery. However, little is known so far on the consequences of a treatment of brain cells with such nanoparticles. In order to study the biocompatibility of iron oxide nanoparticles and the accumulation of iron from such particles in brain cells, we have used astrocyte-rich primary cultures as model system. Iron oxide nanoparticles were chemically synthesized with a yield of about 80% regarding the iron content. Transmission electron microscopy revealed that the synthesized iron oxide nanoparticles had a diameter of about 10 nm. Exposure of astrocyte cultures to 100 microM iron that were supplied as citrate-coated iron oxide nanoparticles did not cause any acute loss in cell viability but resulted in an almost linear increase in the cellular iron content that reached after 6 h of incubation at 37 degrees C a total cellular iron content of about 300 nmol/mg protein. The rate of iron accumulation from iron oxide nanoparticles was significantly higher than that from the low molecular weight iron complex ferric ammonium citrate (FAC). Lowering the incubation temperature from 37 degrees C to 4 degrees C reduced the iron accumulation rate from iron oxide nanoparticles or FAC by about 60%. In addition, presence of ferric or ferrous iron chelators did not affect the cellular iron accumulation from iron oxide nanoparticles. These data suggest that cultured astrocytes are able to accumulate intact iron oxide nanoparticles.

Formation of a Three-Dimensional Multicellular Assembly Using Magnetic Patterning

We demonstrate a facile approach to design three-dimensional cellular assembly of tunable size and controlled geometry with applications for tissue engineering. Three-dimensional cell patterning was performed using external magnetic forces, without the need for substrate chemical or physical modifications. Human endothelial progenitor cells and mouse macrophages were magnetically labeled using anionic citrate-coated iron oxide nanoparticles. Two magnetic tips were designed, and their magnetic field cartographies were calibrated. The focalized magnetic force generated ensured an efficient entrapment of the cells at the tips vicinity. By tuning the magnetic field gradient geometry and intensity, the magnetic cellular load, and the number of cells, we fully described the formation of the three-dimensional multicellular assemblies, and estimated the corresponding packing factor for a large range of experimental conditions.

Images in cardiovascular medicine. Avoiding papillary muscle infarction with myocardial contrast echocardiographic guidance of nonsurgical septal reduction therapy for hypertrophic obstructive cardiomyopathy.

Intracoronary myocardial contrast echocardiography (MCE) can be used to guide the delivery of ethanol during nonsurgical septal reduction therapy for hypertrophic obstructive cardiomyopathy. The echocardiographic contrast agent is injected immediately before the injection of ethanol, down the lumen of the inflated balloon that resides in the target septal coronary artery. This is the artery that supplies the area of the septum involved with the mitral leaflet(s) in causing dynamic outflow tract obstruction. MCE provides a direct visualization of the myocardial territory that should receive ethanol and consequently undergo infarction. In rare instances, the cannulated vessel does not supply the culprit septal segment(s) but supplies instead …