Classification of Magnetic Nanoparticle Systems--Synthesis, Standardization and Analysis Methods in the NanoMag Project.

This study presents classification of different magnetic single- and multi-core particle systems using their measured dynamic magnetic properties together with their nanocrystal and particle sizes. The dynamic magnetic properties are measured with AC (dynamical) susceptometry and magnetorelaxometry and the size parameters are determined from electron microscopy and dynamic light scattering. Using these methods, we also show that the nanocrystal size and particle morphology determines the dynamic magnetic properties for both single- and multi-core particles. The presented results are obtained from the four year EU NMP FP7 project, NanoMag, which is focused on standardization of analysis methods for magnetic nanoparticles.

Magnetorelaxometry assisting biomedical applications of magnetic nanoparticles.

Due to their biocompatibility and small size, iron oxide magnetic nanoparticles (MNP) can be guided to virtually every biological environment. MNP are susceptible to external magnetic fields and can thus be used for transport of drugs and genes, for heat generation in magnetic hyperthermia or for contrast enhancement in magnetic resonance imaging of biological tissue. At the same time, their magnetic properties allow one to develop sensitive and specific measurement methods to non-invasively detect MNP, to quantify MNP distribution in tissue and to determine their binding state. In this article, we review the application of magnetorelaxometry (MRX) for MNP detection. The underlying physical properties of MNP responsible for the generation of the MRX signal with its characteristic parameters of relaxation amplitude and relaxation time are described. Existing single and multi-channel MRX devices are reviewed. Finally, we thoroughly describe some applications of MRX to cellular MNP quantification, MNP organ distribution and MNP-based binding assays. Providing specific MNP signals, a detection limit down to a few nanogram MNP, in-vivo capability in conscious animals and measurement times of a few seconds, MRX is a valuable tool to improve the application of MNP for diagnostic and therapeutic purposes.

Experimental demonstration of improved magnetorelaxometry imaging performance using optimized coil configurations.

BACKGROUND: Magnetorelaxometry imaging is an experimental imaging technique capable of reconstructing magnetic nanoparticle distributions inside a volume noninvasively and with high specificity. Thus, magnetorelaxometry imaging is a promising candidate for monitoring a number of therapeutical approaches that employ magnetic nanoparticles, such as magnetic drug targeting and magnetic hyperthermia, to guarantee their safety and efficacy. Prior to a potential clinical application of this imaging modality, it is necessary to optimize magnetorelaxometry imaging systems to produce reliable imaging results and to maximize the reconstruction accuracy of the magnetic nanoparticle distributions. Multiple optimization approaches were already applied throughout a number of simulation studies, all of which yielded increased imaging qualities compared to intuitively designed measurement setups. PURPOSE: None of these simulative approaches was conducted in practice such that it still remains unclear if the theoretical results are achievable in an experimental setting. In this study, we demonstrate the technical feasibility and the increased reconstruction accuracy of optimized coil configurations in two distinct magnetorelaxometry setups. METHODS: The electromagnetic coil positions and radii of a cuboidal as well as a cylindrical magnetorelaxometry imaging setup are optimized by minimizing the system matrix condition numbers of their corresponding linear forward models. The optimized coil configurations are manufactured alongside with two regular coil grids. Magnetorelaxometry measurements of three cuboidal and four cylindrical magnetic nanoparticle phantoms are conducted, and the resulting reconstruction qualities of the optimized and the regular coil configurations are compared. RESULTS: The computed condition numbers of the optimized coil configurations are approximately one order of magnitude lower compared to the regular coil grids. The reconstruction results show that for both setups, every phantom is recovered more accurately by the optimized coil configurations compared to the regular coil grids. Additionally, the optimized coil configurations yield better signal qualities. CONCLUSIONS: The presented experimental study provides a proof of the practicality and the efficacy of optimizing magnetorelaxometry imaging systems with respect to the condition numbers of their system matrices, previously only demonstrated in simulations. From the promising results of our study, we infer that the minimization of the system matrix condition number will also enable the practical optimization of other design parameters of magnetorelaxometry imaging setups (e.g., sensor configuration, coil currents, etc.) in order to improve the achievable reconstruction qualities even further, eventually paving the way towards clinical application of this imaging modality.

Whither Magnetic Hyperthermia? A Tentative Roadmap.

The scientific community has made great efforts in advancing magnetic hyperthermia for the last two decades after going through a sizeable research lapse from its establishment. All the progress made in various topics ranging from nanoparticle synthesis to biocompatibilization and in vivo testing have been seeking to push the forefront towards some new clinical trials. As many, they did not go at the expected pace. Today, fruitful international cooperation and the wisdom gain after a careful analysis of the lessons learned from seminal clinical trials allow us to have a future with better guarantees for a more definitive takeoff of this genuine nanotherapy against cancer. Deliberately giving prominence to a number of critical aspects, this opinion review offers a blend of state-of-the-art hints and glimpses into the future of the therapy, considering the expected evolution of science and technology behind magnetic hyperthermia.

Quantification of specific bindings of biomolecules by magnetorelaxometry.

The binding reaction of the biomolecules streptavidin and anti-biotin antibody, both labelled by magnetic nanoparticles (MNP), to biotin coated on agarose beads, was quantified by magnetorelaxometry (MRX). Highly sensitive SQUID-based MRX revealed the immobilization of the MNP caused by the biotin-streptavidin coupling. We found that about 85% of streptavidin-functionalised MNP bound specifically to biotin-agarose beads. On the other hand only 20% of antibiotin-antibody functionalised MNP were specifically bound. Variation of the suspension medium revealed in comparison to phosphate buffer with 0.1% bovine serum albumin a slight change of the binding behaviour in human serum, probably due to the presence of functioning (non heated) serum proteins. Furthermore, in human serum an additional non-specific binding occurs, being independent from the serum protein functionality.The presented homogeneous bead based assay is applicable in simple, uncoated vials and it enables the assessment of the binding kinetics in a volume without liquid flow. The estimated association rate constant for the MNP-labelled streptavidin is by about two orders of magnitude smaller than the value reported for free streptavidin. This is probably due to the relatively large size of the magnetic markers which reduces the diffusion of streptavidin. Furthermore, long time non-exponential kinetics were observed and interpreted as agglutination of the agarose beads.

Improved sensitivity and limit-of-detection using a receive-only coil in magnetic particle imaging.

Magnetic particle imaging (MPI) is an imaging modality capable of quantitatively determining the 3D distribution of a magnetic nanoparticle (MNP) ensemble. In this work, we present a method for reducing the MNP limit of detection by employing a new receive-only coil (Rx-coil) for signal acquisition. The new signal detector is designed to improve the sensitivity and thus quality of reconstructed images. We present characterization measurements conducted with the prototype Rx-coil installed in a preclinical MPI scanner. The gradiometric design of the Rx-coil attenuates the unwanted signal contributions arising from the excitation field, leading to a 17 dB lower background level compared to the conventional dual-purpose coil (TxRx-coil), which is crucial for detecting low amounts of MNP. Network analyzer measurements of the frequency-dependent coil sensitivity, as well as spectral analysis of recorded MPI data demonstrate an overall increase of the coil sensitivity of about +12 dB for the Rx-coil. Comparisons of the sensitivity distributions revealed no significant degradations in terms of homogeneity for the Rx-coil compared to the TxRx-coil in an imaging volume of 6 × 3 × 3 cm3. Finally, the limit of detection was determined experimentally for each coil type using a serial dilution of MNPs, resulting in values of 133 ng of iron for the conventional TxRx-coil and 20 ng for the new Rx-coil, using an acquisition time of 2 s. A linear relationship between the reconstructed signal intensities and the iron mass in the samples was observed with coefficients of determination (R2) of above 99% in the range of the limit of detection to 3 103ng(Fe). These results open the way for improved image quality and faster acquisition time in pre-clinical MPI scanners.

Comparison of magnetocardiography and electrocardiography.

OBJECTIVE: Automated techniques were developed for the measurement of cardiac repolarisation using magnetocardiography. METHODS: This was achieved by collaboration with the Physikalisch-Technische Bundesanstalt (PTB), Berlin, Germany and the Grönemeyer Institute of Microtherapy, Bochum, Germany, to obtain recordings of magnetocardiograms (MCGs) in cardiac patients and healthy subjects. Manual and automated ventricular repolarisation measurements from MCGs were evaluated to determine the clinical relevance of these measurements compared with electrocardiograms (ECGs). RESULTS: Results showed that MCG and ECG T-wave shapes differed and that manual repolarisation measurement was significantly influenced by T-wave amplitude. Automatic measurements of repolarisation in both MCGs and ECGs differed between techniques. The effects of filtering on the waveforms showed that filtering in some MCG research systems could significantly influence the results, with 20 ms differences common. In addition, MCGs were better able to identify differences in the distribution of cardiac magnetic field strength during repolarisation and depolarisation between normal subjects and cardiac patients. Differences were also determined in ventricular repolarisation between MCGs and ECGs, which cannot be explained by channel/lead numbers or amplitude effects alone. CONCLUSION: The techniques developed are essential, because of the many extra MCG channels to analyse, and will encourage the use of MCG facilities.

Magnetorelaxometry procedures for quantitative imaging and characterization of magnetic nanoparticles in biomedical applications.

BACKGROUND: Quantitative knowledge about the spatial distribution and local environment of magnetic nanoparticles (MNPs) inside an organism is essential for guidance and improvement of biomedical applications such as magnetic hyperthermia and magnetic drug targeting. Magnetorelaxometry (MRX) provides such quantitative information by detecting the magnetic response of MNPs following a fast change in the applied magnetic field. METHODS: In this article, we review our MRX based procedures that enable both the characterization and the quantitative imaging of MNPs in a biomedical environment. RESULTS: MRX characterization supported the selection of an MNP system with colloidal stability and suitable cellular MNP uptake. Spatially resolved MRX, a procedure employing multi-channel MRX measurements allowed for in-vivo monitoring of the MNP distribution in a pre-clinical carcinoma animal model. Extending spatially resolved MRX by consecutive magnetization of distinct parts of the sample led to a demonstration of MRX tomography. With this tomography, we reconstructed the three dimensional MNP distribution inside animal sized phantoms with a sensitivity of milligrams of MNPs per cm3. In addition, the targeting efficiency of MNPs in whole blood was assessed using a flow phantom and MRX quantification. CONCLUSION: These MRX based measurement and analysis procedures have substantially supported the development of MNP based biomedical applications.

Fetal magnetocardiography measurements with an array of microfabricated optically pumped magnetometers.

Following the rapid progress in the development of optically pumped magnetometer (OPM) technology for the measurement of magnetic fields in the femtotesla range, a successful assembly of individual sensors into an array of nearly identical sensors is within reach. Here, 25 microfabricated OPMs with footprints of 1 cm(3) were assembled into a conformal array. The individual sensors were inserted into three flexible belt-shaped holders and connected to their respective light sources and electronics, which reside outside a magnetically shielded room, through long optical and electrical cables. With this setup the fetal magnetocardiogram of a pregnant woman was measured by placing two sensor belts over her abdomen and one belt over her chest. The fetal magnetocardiogram recorded over the abdomen is usually dominated by contributions from the maternal magnetocardiogram, since the maternal heart generates a much stronger signal than the fetal heart. Therefore, signal processing methods have to be applied to obtain the pure fetal magnetocardiogram: orthogonal projection and independent component analysis. The resulting spatial distributions of fetal cardiac activity are in good agreement with each other. In a further exemplary step, the fetal heart rate was extracted from the fetal magnetocardiogram. Its variability suggests fetal activity. We conclude that microfabricated optically pumped magnetometers operating at room temperature are capable of complementing or in the future even replacing superconducting sensors for fetal magnetocardiography measurements.

Characterization of magnetic nanoparticle systems with respect to their magnetic particle imaging performance.

The optimization of magnetic nanoparticles (MNPs) as markers for magnetic particle imaging (MPI) requires an understanding of the relationship between the harmonics spectrum and the structural and magnetic properties of the MNPs. Although magnetic particle spectroscopy (MPS) - carried out at the same excitation frequency as the given MPI system - represents a straightforward technique to study MNPs for their suitability for MPI, a complete understanding of the mechanisms and differences between different tracer materials requires additional measurements of the static and dynamic magnetic behavior covering additional field and time ranges. Furthermore, theoretical models are needed, which correctly account for the static and dynamic magnetic properties of the markers. In this paper, we give an overview of currently used theoretical models for the explanation of amplitude and phase of the harmonics spectra as well as of the various static and dynamic magnetic techniques, which are applied for the comprehensive characterization of MNPs for MPI. We demonstrate on two multicore MNP model systems, Resovist(®) and FeraSpin™ Series, how a detailed picture of the MPI performance can be obtained by combining various static and dynamic magnetic measurements.

Magnetic nanoparticle imaging by means of minimum norm estimates from remanence measurements.

In magnetic nanoparticle imaging, magnetic nanoparticles are coated and functionalized to bind to specific targets. After measuring their magnetic relaxation or remanence, their distribution can be determined by means of inverse methods. The reconstruction algorithm presented in this paper includes first a dipole fit using a Levenberg-Marquardt optimizer to determine the reconstruction plane. Secondly, a minimum norm estimate is obtained on a regular grid placed in that plane. Computer simulations involving different parameter sets and conditions show that the used approach allows for the reconstruction of distributed sources, although the reconstructed shapes are distorted by blurring effects. The reconstruction quality depends on the signal-to-noise ratio of the measurements and decreases with larger sensor-source distances and higher grid spacings. In phantom measurements, the magnetic remanence of nanoparticle columns with clinical relevant sizes is determined with two common measurement systems. The reconstructions from these measurements indicate that the approach is applicable for clinical measurements. Our results provide parameter sets for successful application of minimum norm approaches to Magnetic Nanoparticle Imaging.

Pseudo current density maps of electrophysiological heart, nerve or brain function and their physical basis.

BACKGROUND: In recent years the visualization of biomagnetic measurement data by so-called pseudo current density maps or Hosaka-Cohen (HC) transformations became popular. METHODS: The physical basis of these intuitive maps is clarified by means of analytically solvable problems. RESULTS: Examples in magnetocardiography, magnetoencephalography and magnetoneurography demonstrate the usefulness of this method. CONCLUSION: Hardware realizations of the HC-transformation and some similar transformations are discussed which could advantageously support cross-platform comparability of biomagnetic measurements.

Comparison of magnetocardiography and electrocardiography: a study of automatic measurement of dispersion of ventricular repolarization.

AIMS: There is some dispute over the clinical significance of dispersion of ventricular repolarization measurements from the electrocardiogram. Recent studies have indicated that multichannel magnetocardiograms (MCGs), which non-invasively measure cardiac magnetic field strength from many sites above the body surface, may provide independent information from ECGs about ventricular repolarization dispersion. For this study, magnetocardiography and electrocardiography were compared from automatic measurements of dispersion of ventricular repolarization. METHODS AND RESULTS: Dispersion of ventricular repolarization time was determined in MCGs and standard ECGs recorded simultaneously from 27 healthy volunteers and 22 cardiac patients. Two automatic techniques were used to determine the interval of ventricular repolarization. There were significant differences in ventricular dispersion between ECG and MCG measurements, with multichannel MCG greater than ECG by 52 (47) ms [mean (SD)] (P<0.00001) and 12-channel MCG greater by 17 (40) ms (P<0.004) across techniques and all subjects. Magnetocardiograms had the greater discriminating power between normal and cardiac patients with differences of 46 (18) ms (P<0.017) for multichannel MCG and 44 (16) ms (P<0.005) for 12-channel MCG, compared with 16 (7) ms (P<0.04) for ECG. CONCLUSION: Magnetocardiography has the power to discriminate regional cardiac conduction differences.

MCG to ECG source differences: measurements and a two-dimensional computer model study.

In this work we combine body surface potential map (BSPM) and magnetocardiogram (MCG) measurements with computer simulations in order to elucidate a recent thesis that claims the orthogonality of the main sources of MCG and ECG. Body surface currents and MCG pseudo currents are calculated from measured BSPM and MCG data, respectively. In contrast to the MCG-ECG source orthogonality thesis, we observe the main orientation of the BSPM currents and MCG pseudo currents to have similar axis during most of the depolarization R wave. In an attempt to explain such measurements we simulate a 2D transmural slice of the left ventricle in contact to a volume conductor. The main magnetic source currents along the wave front are indeed orthogonal to the extracellular electric current. However, fiber orientation inhomogeneity through the ventricular wall, volume conductor interface, wave front shape and extra- and intracellular potential distributions, all distort the symmetry of the current loops that contain the wave front currents. The resulting asymmetry rotates the main axis of the pseudo MCG currents away from the orthogonal axis of the body surface currents. Thus, the simulation results could solve the apparent contradiction between the orthogonal source theory and the observed similar ECG and MCG main current axis.

Errors in repolarization measurement using magnetocardiography.

Multichannel magnetocardiography (MCG) noninvasively measures variations in magnetic field strength from many sites at the body surface, potentially providing useful regional information about ventricular repolarization. MCGs contain features similar to ECGs, and although errors associated with repolarization measurement have been quantified for ECGs, no comparative data exists for MCGs. In this study, errors in manual measurement of repolarization interval in the MCG were determined. Sixteen MCG channels and three ECG leads were recorded simultaneously in eight healthy subjects. Each recording was displayed in a random order on a computer screen, in presentations with different noise levels, time display widths, and amplitude display heights. In total, manual measurement of repolarization intervals in 2,048 (eight subjects x 16 channels x eight presentations x two repeats) MCGs were made by each of four analysts. Measured repolarization intervals were reduced by 3 ms when noise was added and by a further 3 ms when this noise was doubled. Intervals were shortened by 9 ms when the time display width was doubled and by a further 10 ms when the display width was doubled again. Measurements increased by 7 ms for a doubling of amplitude display height, equivalent to a doubling of T wave height. There were also consistent differences between analysts; amounting to a greatest mean difference of 24 ms. Display characteristics, added noise, and different analysts thus affect manual repolarization interval measurements in MCG. The errors detected demonstrate the importance of a standard presentation for repolarization measurement in the MCG.

Comparison of automatic repolarization measurement techniques in the normal magnetocardiogram.

Multichannel MCG noninvasively measures cardiac magnetic field strength from many sites at the body surface, potentially providing useful regional information about ventricular repolarization. Previous work on ECGs has shown that automatic techniques for repolarization measurement are better than manual measurement at discriminating patients with cardiac conditions from normal subjects. Although automatic repolarization measurement techniques have been quantified for ECGs, no comparative data exists for the MCG. In this study four different automatic repolarization (QT) interval techniques for detecting T wave end in the MCG were compared. The influence of MCG filtering on the automatic algorithms was also quantified. MCGs were obtained at 49 sites over the heart from 23 normal subjects. Automatic measurements of the repolarization (QT) interval were made following the addition of different high pass (0.25, 0.5, 1 Hz) and low pass (100, 60, 40, 30 Hz) filters. There were consistent differences between automatic techniques in the unfiltered data amounting to greatest mean difference of 52.3 ms. Low pass filtering significantly increased the automatic repolarization (QT) interval relative to unfiltered measurement by 6.5 (3.2) ms (mean SD) for 100 Hz, 6.0 (3.0) ms for 60 Hz, 8.1 (3.2) ms for 40 Hz, and 8.8 (3.1) ms for 30 Hz across all techniques. High pass filtering significantly decreased the value by -2.6 (6.0) ms for 0.25 Hz, -5.5 (5.3) ms for 0.5 Hz, and -17.1 (7.8) ms for 1 Hz. Automatic measurements of repolarization (QT) in the MCG differ between techniques and are influenced by filtering. These effects should be considered when comparing results.

Magnetocardiography for pharmacology safety studies requiring high patient throughput and reliability.

Recent guideline drafts of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) underline the necessity to test nonantiarrhythmic drugs for their potential to prolong the QT or the corrected QT (QTc) interval. The implementation of these guidelines requires a large amount of ECG measurements on animals and humans in preclinical and clinical phases of the drug development process. We propose the use of magnetocardiography (MCG) as a complementary method with particular advantages in high-throughput studies, where signal quality and reliability are key factors. Our proposal is based on a review of recent MCG studies investigating the repolarization phase and results of methodological work assessing QT interval parameters from the MCG. The applicability of MCG for pre-clinical in-vivo studies is demonstrated by the ease of measurement in unrestrained non-anesthetized rabbits, guinea pigs, and hamsters..