Hamed Arami

Targeting of primary breast cancers and metastases in a transgenic mouse model using rationally designed multifunctional SPIONs.

Breast cancer remains one of the most prevalent and lethal malignancies in women. The inability to diagnose small volume metastases early has limited effective treatment of stage 4 breast cancer. Here we report the rational development and use of a multifunctional superparamagnetic iron oxide nanoparticle (SPION) for targeting metastatic breast cancer in a transgenic mouse model and imaging with magnetic resonance (MR). SPIONs coated with a copolymer of chitosan and polyethylene glycol (PEG) were labeled with a fluorescent dye for optical detection and conjugated with a monoclonal antibody against the neu receptor (NP-neu). SPIONs labeled with mouse IgG were used as a nontargeting control (NP-IgG). These SPIONs had desirable physiochemical properties for in vivo applications such as near neutral zeta potential and hydrodynamic size around 40 nm and were highly stable in serum containing medium. Only NP-neu showed high uptake in neu expressing mouse mammary carcinoma (MMC) cells which was reversed by competing free neu antibody, indicating their specificity to the neu antigen. In vivo, NP-neu was able to tag primary breast tumors and significantly, only NP-neu bound to spontaneous liver, lung, and bone marrow metastases in a transgenic mouse model of metastatic breast cancer, highlighting the necessity of targeting for delivery to metastatic disease. The SPIONs provided significant contrast enhancement in MR images of primary breast tumors; thus, they have the potential for MRI detection of micrometastases and provide an excellent platform for further development of an efficient metastatic breast cancer therapy.

In vivo delivery, pharmacokinetics, biodistribution and toxicity of iron oxide nanoparticles

Iron oxide nanoparticles (IONPs) have been extensively used during the last two decades, either as effective bio-imaging contrast agents or as carriers of biomolecules such as drugs, nucleic acids and peptides for controlled delivery to specific organs and tissues. Most of these novel applications require elaborate tuning of the physiochemical and surface properties of the IONPs. As new IONPs designs are envisioned, synergistic consideration of the bodys innate biological barriers against the administered nanoparticles and the short and long-term side effects of the IONPs become even more essential. There are several important criteria (e.g. size and size-distribution, charge, coating molecules, and plasma protein adsorption) that can be effectively tuned to control the in vivo pharmacokinetics and biodistribution of the IONPs. This paper reviews these crucial parameters, in light of biological barriers in the body, and the latest IONPs design strategies used to overcome them. A careful review of the long-term biodistribution and side effects of the IONPs in relation to nanoparticle design is also given. While the discussions presented in this review are specific to IONPs, some of the information can be readily applied to other nanoparticle systems, such as gold, silver, silica, calcium phosphates and various polymers.

Magnetic Particle Imaging With Tailored Iron Oxide Nanoparticle Tracers

Magnetic particle imaging (MPI) shows promise for medical imaging, particularly in angiography of patients with chronic kidney disease. As the first biomedical imaging technique that truly depends on nanoscale materials properties, MPI requires highly optimized magnetic nanoparticle tracers to generate quality images. Until now, researchers have relied on tracers optimized for MRI T2*-weighted imaging that are sub-optimal for MPI. Here, we describe new tracers tailored to MPIs unique physics, synthesized using an organic-phase process and functionalized to ensure biocompatibility and adequate in vivo circulation time. Tailored tracers showed up to 3 × greater signal-to-noise ratio and better spatial resolution than existing commercial tracers in MPI images of phantoms.

Cell transcytosing poly-arginine coated magnetic nanovector for safe and effective siRNA delivery

Lack of safe and effective carriers for delivery of RNA therapeutics remains a barrier to its broad clinical application. We report the development of a cell tanscytosing magnetic nanovector engineered as an siRNA carrier. Iron oxide nanoparticles were modified with poly(ethylene glycol) (PEG), small interfering RNA (siRNA), and a cationic polymer layer. Three nanovector formulations with cationic polymer coatings of poly-arginine (pArg), polylysine (pLys), and polyethylenimine (PEI), respectively, were prepared. The three nanovector formulations where evaluated for safety and ability to promote gene silencing in three types of cancer cells C6/GFP(+), MCF7/GFP(+), and TC2/GFP(+), mimicking human cancers of the brain, breast, and prostate, respectively. Cell viability and fluorescence quantification assays revealed that pArg-coated nanovectors were most effective in promoting gene knockdown and least toxic of the three nanovector formulations tested. Transmission electron microscopy (TEM) imaging of nanovector treated cells further demonstrated that pArg-coated nanovectors enter cells through cell transcytosis, while pLys and PEI coated nanovectors enter cells endocytosis. Our findings suggest that NPs engineered to exploit the cell transcytosis intracellular trafficking pathway may offer a more safe and efficient route for siRNA delivery.

Chitosan-Coated Iron Oxide Nanoparticles for Molecular Imaging and Drug Delivery

Iron oxide nanoparticles (IONPs) are a new class of nanomaterials which have attracted extensive interest for application in in vivo magnetic resonance imaging (MRI) due to their intrinsic superparamagnetic and biodegradable properties. Performance of the IONPs is largely dependent upon the properties of their surface coatings, which serve to prevent nanoparticle agglomeration, reduce the risk of immunogenicity, and limit nonspecific cellular uptake. Among the coating materials studied to date, chitosan has drawn considerable attention. Commonly derived from crustacean shells, chitosan is a natural linear polysaccharide and has ample reactive functional groups that can serve as anchors for conjugation of therapeutics, targeting ligands, and imaging agents. Because of these unique attributes, chitosan-coated IONPs are becoming more desirable for cancer imaging and therapy applications. This chapter discusses the current advances and challenges in synthesis of chitosan-coated IONPs, and their subsequent surface modifications for applications in cancer diagnosis and therapy.

Tailoring the magnetic and pharmacokinetic properties of iron oxide magnetic particle imaging tracers

Abstract Magnetic particle imaging (MPI) is an attractive new modality for imaging distributions of iron oxide nanoparticle tracers in vivo. With exceptional contrast, high sensitivity, and good spatial resolution, MPI shows promise for clinical imaging in angiography and oncology. Critically, MPI requires high-quality iron oxide nanoparticle tracers with tailored magnetic and surface properties to achieve its full potential. In this review, we discuss optimizing iron oxide nanoparticles’ physical, magnetic, and pharmacokinetic properties for MPI, highlighting results from our recent work in which we demonstrated tailored, biocompatible iron oxide nanoparticle tracers that provided two times better linear spatial resolution and five times better signal-to-noise ratio than Resovist.

Quantitative "Hot Spot" Imaging of Transplanted Stem Cells using Superparamagnetic Tracers and Magnetic Particle Imaging (MPI).

Magnetic labeling of stem cells enables their noninvasive detection by magnetic resonance imaging (MRI). In practical terms, most MRI studies have been limited to the visualization of local engraftment because other sources of endogenous hypointense contrast complicate the interpretation of systemic (whole-body) cell distribution. In addition, MRI cell tracking is inherently nonquantitative in nature. We report herein on the potential of magnetic particle imaging (MPI) as a novel tomographic technique for noninvasive “hot-spot” imaging and quantification of stem cells using superparamagnetic iron oxide (SPIO) tracers. Neural and mesenchymal stem cells, representing small and larger cell bodies, were labeled with 3 different SPIO tracer formulations, including 2 preparations (Feridex and Resovist) that have previously been used in clinical MRI cell-tracking studies. Magnetic particle spectroscopy measurements demonstrated a linear correlation between MPI signal and iron content for both free particles in homogeneous solution and for internalized and aggregated particles in labeled cells over a wide range of concentrations. The overall MPI signal ranged from 1 × 10−3 to 3 × 10−4 Am2/g Fe, which was equivalent to 2 × 10−14 to 1 × 10−15 Am2 per cell, indicating that cell numbers can be quantified with MPI analogous to the use of radiotracers in nuclear medicine or fluorine tracers in 19F MRI. When SPIO-labeled cells were transplanted in the mouse brain, they could be readily detected by MPI at a detection threshold of about 5 × 104 cells, with MPI/MRI overlays showing an excellent agreement between the hypointense MRI areas and MPI hot spots. The calculated tissue MPI signal ratio for 100 000 vs 50 000 implanted cells was 2.08. Hence, MPI can potentially be further developed for quantitative and easy-to-interpret, tracer-based noninvasive cell imaging, preferably with MRI as an adjunct anatomical imaging modality.

Powder Metallurgical Fabrication and Characterization of Nanostructured Porous NiTi Shape-Memory Alloy

Production of NiTi alloy from elemental powders was conducted by mechanical alloying (MA) and sintering of the raw materials. Effects of milling time and milling speed (RPM) on crystallite size, lattice strain, and XRD peak intensities were investigated by X-ray analysis of the alloy. Powder compaction and sintering time and temperature effects on pore percentage of the as-mixed and the mechanically alloyed samples were empirically evaluated. The crystallite size of the mechanically alloyed Ni50Ti50 samples decreased with MA duration and with the milling speed. Depending on the crystal structure of the raw materials, the lattice strain increases with the milling duration. Metallographic studies proved the existence of martensitic B19′ after sintering of both the as-mixed and the mechanically alloyed samples. Its amount was, however, greater for the former. Sintering lowered the porosity of the samples; no matter what powder (as-mixed or mechanically alloyed) was used. The porosity was greater, however, for the MA powders. This difference seemed to be due to the sharper liquid phase sintering effect of the as-mixed samples.

Lactoferrin conjugated iron oxide nanoparticles for targeting brain glioma cells in magnetic particle imaging.

Magnetic Particle Imaging (MPI) is a new real-time imaging modality, which promises high tracer mass sensitivity and spatial resolution directly generated from iron oxide nanoparticles. In this study, monodisperse iron oxide nanoparticles with median core diameters ranging from 14 to 26 nm were synthesized and their surface was conjugated with lactoferrin to convert them into brain glioma targeting agents. The conjugation was confirmed with the increase of the hydrodynamic diameters, change of zeta potential, and Bradford assay. Magnetic particle spectrometry (MPS), performed to evaluate the MPI performance of these nanoparticles, showed no change in signal after lactoferrin conjugation to nanoparticles for all core diameters, suggesting that the MPI signal is dominated by Néel relaxation and thus independent of hydrodynamic size difference or presence of coating molecules before and after conjugations. For this range of core sizes (14-26 nm), both MPS signal intensity and spatial resolution improved with increasing core diameter of nanoparticles. The lactoferrin conjugated iron oxide nanoparticles (Lf-IONPs) showed specific cellular internalization into C6 cells with a 5-fold increase in MPS signal compared to IONPs without lactoferrin, both after 24 h incubation. These results suggest that Lf-IONPs can be used as tracers for targeted brain glioma imaging using MPI.

Room-temperature detection of a single 19 nm super-paramagnetic nanoparticle with an imaging magnetometer

We demonstrate room temperature detection of isolated single 19 nm super-paramagnetic nanoparticles (SPNs) with a wide-field optical microscope platform suitable for biological integration. The particles are made of magnetite (Fe3O4) and are thus non-toxic and biocompatible. Detection is accomplished via optically detected magnetic resonance imaging using nitrogen-vacancy defect centers in diamond, resulting in a DC magnetic field detection limit of 2.4 μT. This marks a large step forward in the detection of SPNs, and we expect that it will allow for the development of magnetic-field-based biosensors capable of detecting a single molecular binding event.