Synthesis of Biocompatible Superparamagnetic Iron Oxide Nanoparticles (SPION) under Different Microfluidic Regimes.

Superparamagnetic iron oxide nanoparticles (SPION) have a great potential in both diagnostic and therapeutic applications as they provide contrast in magnetic resonance imaging techniques and allow magnetic hyperthermia and drug delivery. Though various types of SPION are commercially available, efforts to improve the quality of SPION are highly in demand. Here, we describe a strategy for optimization of SPION synthesis under microfluidics using the coprecipitation approach. Synthesis parameters such as temperature, pH, iron salt concentration, and coating materials were investigated in continuous and segmented flows. Continuous flow allowed synthesizing particles of a smaller size and higher stability than segmented flow, while both conditions improved the quality of particles compared to batch synthesis. The most stable particles were obtained at a synthesis condition of 6.5 M NH4OH base, iron salt (Fe2+/Fe3+) concentration ratio of 4.3/8.6, carboxymethyl dextran coating of 20 mg/mL, and temperature of 70 °C. The synthesized SPION exhibited a good efficiency in labeling of human platelets and did not impair cells. Our study under flow conditions provides an optimal protocol for the synthesis of better and biocompatible SPION that contributes to the development of nanoparticles for medical applications.

Elastic transformation of histological slices allows precise co-registration with microCT data sets for a refined virtual histology approach.

Although X-ray based 3D virtual histology is an emerging tool for the analysis of biological tissue, it falls short in terms of specificity when compared to conventional histology. Thus, the aim was to establish a novel approach that combines 3D information provided by microCT with high specificity that only (immuno-)histochemistry can offer. For this purpose, we developed a software frontend, which utilises an elastic transformation technique to accurately co-register various histological and immunohistochemical stainings with free propagation phase contrast synchrotron radiation microCT. We demonstrate that the precision of the overlay of both imaging modalities is significantly improved by performing our elastic registration workflow, as evidenced by calculation of the displacement index. To illustrate the need for an elastic co-registration approach we examined specimens from a mouse model of breast cancer with injected metal-based nanoparticles. Using the elastic transformation pipeline, we were able to co-localise the nanoparticles to specifically stained cells or tissue structures into their three-dimensional anatomical context. Additionally, we performed a semi-automated tissue structure and cell classification. This workflow provides new insights on histopathological analysis by combining CT specific three-dimensional information with cell/tissue specific information provided by classical histology.

Ex vivo Live Cell Imaging of Nanoparticle-Cell Interactions in the Mouse Lung.

A successful clinical translation of novel nanoparticle-based cancer therapeutics requires a thorough preclinical investigation of their interaction with immune, tumor and endothelial cells as well as components of the tumor-microenvironment. Although high-resolution microscopy images of fixed tumor tissue specimens can provide valuable information in this regard, they are only static snapshots of a momentary event. Here we describe a superior alternative fluorescence microscopy approach to assess the feasibility of investigating nanoparticle-cell interactions in the mouse lung live and over time at nanometer resolution. We applied fluorescent lung tumor cells and Barium-based fluorescently labeled nanoparticles to nude mice or to CD68-EGFP transgenic mice for visualization of the monocyte-macrophage lineage. Shortly before imaging, fluorescently labeled lectin was intravenously injected for staining of the blood vessels. The lung was filled ex vivo with 1% agarose and individual lung lobes were imaged over time using a confocal microscope with Airyscan technology. Time series demonstrate that live cell imaging of lung lobes can be performed for at least 4 h post mortem. Time-lapse movies illustrate the dynamics of the nanoparticles within the pulmonary circulation and their uptake by immune cells. Moreover, the exchange of nanoparticle material between cancer cells was observed over time. Fluorescent monocytes in lungs of CD68-EGFP transgenic mice could be visualized within blood vessels in the process of interaction with tumor cells and nanoparticles. This high resolution ex vivo live cell imaging approach provides an excellent 4D tool to obtain valuable information on the behavior of tumor and immune cells at first encounter with nanoparticles and may contribute to the understanding of how nanoparticles interact with cells supporting the development of therapeutic strategies based on nanoparticulate drug delivery systems.

Magnetite-Arginine Nanoparticles as a Multifunctional Biomedical Tool.

Iron oxide nanoparticles are a promising platform for biomedical applications, both in terms of diagnostics and therapeutics. In addition, arginine-rich polypeptides are known to penetrate across cell membranes. Here, we thus introduce a system based on magnetite nanoparticles and the polypeptide poly-l-arginine (polyR-Fe3O4). We show that the hybrid nanoparticles exhibit a low cytotoxicity that is comparable to Resovist®, a commercially available drug. PolyR-Fe3O4 particles perform very well in diagnostic applications, such as magnetic particle imaging (1.7 and 1.35 higher signal respectively for the 3rd and 11th harmonic when compared to Resovist®), or as contrast agents for magnetic resonance imaging (R2/R1 ratio of 17 as compared to 11 at 0.94 T for Resovist®). Moreover, these novel particles can also be used for therapeutic purposes such as hyperthermia, achieving a specific heating power ratio of 208 W/g as compared to 83 W/g for Feridex®, another commercially available product. Therefore, we envision such materials to play a role in the future theranostic applications, where the arginine ability to deliver cargo into the cell can be coupled to the magnetite imaging properties and cancer fighting activity.

Introducing Specificity to Iron Oxide Nanoparticle Imaging by Combining 57Fe-Based MRI and Mass Spectrometry.

Iron oxide nanoparticles (ION) are highly sensitive probes for magnetic resonance imaging (MRI) that have previously been used for in vivo cell tracking and have enabled implementation of several diagnostic tools to detect and monitor disease. However, the in vivo MRI signal of ION can overlap with the signal from endogenous iron, resulting in a lack of detection specificity. Therefore, the long-term fate of administered ION remains largely unknown, and possible tissue deposition of iron cannot be assessed with established methods. Herein, we combine nonradioactive 57Fe-ION MRI with ex vivo laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) imaging, enabling unambiguous differentiation between endogenous iron (56Fe) and iron originating from applied ION in mice. We establish 57Fe-ION as an in vivo MRI sensor for cell tracking in a mouse model of subcutaneous inflammation and for assessing the long-term fate of 57Fe-ION. Our approach resolves the lack of detection specificity in ION imaging by unambiguously recording a 57Fe signature.

Branch-Like Iron Nitride and Carbide Magnetic Fibres Using an Electrospinning Technique.

Fe3 N and Fe3 C nanocomposites have a wide range of applications thanks to their ceramic nature, magnetic properties, conductivity and catalytic activity, just to cite some. In many fields optimal performances are ensured by crystallinity, homogeneity and hierarchical organization. In the present paper, crystalline, magnetic and well-defined nanofibres of iron nitride and iron carbide/carbon nanocomposite with tunable composition and size were prepared via electrospinning. The starting polymeric material was directly electrospun into fibres and then calcined, leading to a highly homogeneous final product of nanoparticles along the fibres (both outside and inside). A mechanistic study was undertaken and here discussed. The magnetic properties of the as-prepared nanofibres were also studied. The as-prepared final fibre mat composite material can serve as active catalyst, for example, in oxygen reduction reaction (where nanofibres outperformed mere nanoparticles), it can serve as functional support for classical catalytic processes or, thanks to its magnetic properties, can be applied in magnetic-field assisted separation or as magneto-active membranes.

Enhanced Methods to Estimate the Efficiency of Magnetic Nanoparticles in Imaging.

Magnetic resonance imaging (MRI) and magnetic particle imaging (MPI) are powerful methods in the early diagnosis of diseases. Both imaging techniques utilize magnetic nanoparticles that have high magnetic susceptibility, strong saturation magnetization, and no coercivity. FeraSpinTM R and its fractionated products have been studied for their imaging performances; however, a detailed magnetic characterization in their immobilized state is still lacking. This is particularly important for applications in MPI that require fixation of magnetic nanoparticles with the target cells or tissues. We examine the magnetic properties of immobilized FeraSpinTM R, its size fractions, and Resovist®, and use the findings to demonstrate which magnetic properties best predict performance. All samples show some degree of oxidation to hematite, and magnetic interaction between the particles, which impact negatively on image performance of the materials. MRI and MPI performance show a linear dependency on the slope of the magnetization curve, i.e., initial susceptibility, and average blocking temperature. The best performance of particles in immobilized state for MPI is found for particle sizes close to the boundary between superparamagnetic (SP) and magnetically ordered, in which only Néel relaxation is important. Initial susceptibility and bifurcation temperature are the best indicators to predict MRI and MPI performance.

In Vivo Quantification of Myocardial Infarction in Mice Using Micro-CT and a Novel Blood Pool Agent.

We herein developed a micro-CT method using the innovative contrast agent ExiTron™ MyoC 8000 to longitudinally monitor cardiac processes in vivo in small animals. Experiments were performed on healthy mice and mice with myocardial infarction inflicted by ligation of the left anterior descending artery. Time-dependent signal enhancement in different tissues of healthy mice was measured and various contrast agent doses were investigated so as to determine the minimum required dose for imaging of the myocardium. Due to its ability to be taken up by healthy myocardium but not by infarct tissue, ExiTron MyoC 8000 enables detection of myocardial infarction even at a very low dose. The signal enhancement in the myocardium of infarcted mice after contrast agent injection was exploited for quantification of infarct size. The values of infarct size obtained from the imaging method were compared with those obtained from histology and showed a significant correlation (R2 = 0.98). Thus, the developed micro-CT method allows for monitoring of a variety of processes such as cardiac remodeling in longitudinal studies.

Easy access to Ni3N- and Ni-carbon nanocomposite catalysts.

In the search for alternative materials to current expensive catalysts, Ni has been addressed as one of the most promising and, on this trail, its corresponding nitride. However, nickel nitride is a thermally unstable compound, and therefore not easy to prepare especially as nanoparticles. In the present work, a sol-gel-based process (the urea glass route) is applied to prepare well-defined and homogeneous Ni3N and Ni nanoparticles. In both cases, the prepared crystalline nanoparticles (∼25 nm) are dispersed in a carbon matrix forming interesting Ni3N- and Ni-based composites. These nanocomposites were characterised by means of several techniques, such as XRD, HR-TEM, EELS, and the reaction mechanism was investigated by TGA and IR and herein discussed. The catalytic activity of Ni3N is investigated for the first time, to the best of our knowledge, for hydrogenation reactions involving H2, and here compared to the one of Ni. Both materials show good catalytic activities but, interestingly, give a different selectivity between different functional groups (namely, nitro, alkene and nitrile groups).

Stable iron carbide nanoparticle dispersions in [Emim][SCN] and [Emim][N(CN)2] ionic liquids.

Dispersions of Fe(3)C nanoparticles in several ionic liquids (ILs) have been investigated. The ILs are based on 1-ethyl-3-methylimidazolium [Emim] and 1-butyl-3-methylimidazolium [Bmim] cations. Anions are ethylsulfate [ES], methanesulfonate [MS], trifluoromethylsulfonate (triflate) [TfO], tetrafluoroborate [BF(4)], dicyanamide [N(CN)(2)], and thiocyanate [SCN]. Among the ILs studied, [Emim][SCN] and [Emim][N(CN)(2)] stand out because only in these ILs have stable and transparent nanoparticle dispersions been obtained. All other ILs lead to blackish, slightly turbid dispersions or to completely nontransparent suspensions, which often contain undispersed sediment. UV/vis spectroscopy, transmission electron microscopy, and X-ray scattering suggest that the reason for the stabilization of the Fe(3)C nanoparticles in [Emim][SCN] is the leaching of traces of iron from the particles (without affecting the crystal structure of the Fe(3)C particles). The resulting particle surface is thus carbon-rich, which presumably favors the stabilization of the particles. A similar explanation can be postulated for [Emim][N(CN)(2)], with the dicyanamide anion also being a good ligand for iron.