The information about magnetic properties of bone structures in their native state is typically lost during a destructive process of sample preparation for susceptometry by sample reduction to a powder. The low value of the magnetic susceptibility of bone’s mineral component, hydroxyapatite, requires careful calibration of the measuring equipment and process, and introduces the chances of a systematic error. Limited sampling volume of the commercial susceptometers adds to the uncertainty about the magnetic properties of bone structure in the bulk. Taken together, the experimental difficulties of acquiring bone susceptibility data contribute to the scarcity of reliable information about the magnetic properties of bone in literature. We have developed a method that requires minimally destructive sample preparation and in principle can work with the whole bone fragments of small laboratory animals (vertebrae and femurs of laboratory rats). The method is scalable and amenable to modifications of the applied, inhomogeneous magnetic field. This is a report on an exploratory study designed to evaluate the feasibility of the method. It is derived from the first principles in electromagnetics and tested experimentally against measurements using commercial susceptometry.

The purpose of our study was to test the hypothesis that the electromagnetic pulse (EMP) is capable of inducing mechanical vibrations in bone ex vivo . A thin segment of human femur diaphysis (from a tissue repository) suspended on a tensioned line (range $T =$ 2.2–123 N) was exposed to EMP (mean $B =0.64$ T, dB / dt = 5877 T/s, and the mean $B$ -field gradient of 127 T/m) from a solenoid with axis orthogonal to tensioning line, forming a harmonic oscillator whose mechanical vibrations were measured using laser Doppler vibrometry (LDV, noise floor 1 $\mu \text{m}$ /s). Calculated mean Maxwell stress and Lorentz forces acting on a weakly conducting, diamagnetic bone slice point away from the solenoid for maximum sensitivity of LDV measurement. The electromechanical origin of the LDV signal was confirmed by the order-of-magnitude agreement between calculated (range from 12 to 50 $\mu \text{m}$ /s) and measured initial bone velocity amplitudes (e.g., $35.5~\mu \text{m}$ /s ± $7.5~\mu \text{m}$ /s at $T =22.2$ N and $17.7~\mu \text{m}$ /s ± $2.5~\mu \text{m}$ /s at $T =58.2$ N) and the increasing frequency (25–180 Hz) of decaying oscillations with the square root of $T$ over the range of line tensions ( $r^{2} =0.978$ , $p < 10^{-4}$ , and $n =17$ ). Theory and experiment show that magnetic field impulses are capable of exerting measurable mechanical forces on bone ex vivo . The results raise an interesting question if the electromechanical effect could be sufficiently large to contribute to bone remodeling, reportedly sensitive to vibration amplitudes as small as 1 nm, and considering long duration of orthopedic therapy using repetitive EMP (months).

Red blood cell (RBC) magnetophoresis is a function of the concentration of dissolved oxygen (O-2) in normal RBCs measured by cell tracking velocimetry (CTV). The hemoglobin (Hb) magnetic susceptibility changes from diamagnetic (when fully oxygenated, oxyHb) to paramagnetic (when fully deoxygenated, deoxyHb) resulting in a small shift in the normal RBC diamagnetic susceptibility from lower to higher than that of water (-9.05 x 10(-6)). To date, the RBC magnetophoretic motion analyses have been limited to only two quaternary states of the Hb-O-2 complexes, fully oxygenated RBCs (i.e. 100 % O-2 saturation of Hb inside RBCs) and fully deoxygenated RBCs (0 % O-2 saturation of Hb inside RBCs). In this work, the RBC magnetophoretic velocity is measured at small pO(2) intervals for the entire physiological range of 0-160 mmHg. The changes in the RBC magnetophoresis as a function of the O-2 partial pressure in solution (pO(2)) agreed with the theoretical predictions based on the RBC-O-2 equilibrium curve (OEC, measured by Hemox Analyzer and Blood Oxygen Binding System, BOBS (TM)) and the Adair O-2-Hb cooperative binding theory. The results provide a scientific basis for future designs of a "magnetic filter " based on the precise control of the RBC magnetic moment by carefully controlling the pO(2) of the RBC suspension.

Magnetic susceptibility of structurally intact bone may play a role in low frequency (below 10 kHz), low field (below 10 mT amplitude) pulsed electromagnetic field (LF PEMF) treatment of fracture non-unions and an adjunct to cervical fusion surgery, approved by the U.S Food and Drug Administration (FDA). The aim of this study was to determine if a difference in PEMF treatment results in measurable differences in the bone magnetic susceptibility using a model of structurally intact rat vertebrae. We measured bone volume magnetic susceptibility by a custom built mechanical resonator comprising a diametrically-polarized NdFeB permanent dipole magnet cylinder (25.4 x 25.4 mm, height by diameter) rotated about its axis by a stepper motor at even intervals from 53 to 63 RPM, acting as a source of a local, oscillating magnetic field (mean field of 95 mT, gradient of 12 T/m) on an intact laboratory rat lumbar vertebrae (n = 21, from tissue repository from past studies on LF PEMF treatment in vivo) suspended 26.0 mm from magnet axis on a thin line of 80 mm nominal length forming a pendulum of nominal eigenfrequency of 100 Hz. The vertebra motion was captured by a video camera and the geometrical center of its 2D image motion was analyzed by ImagePro (R) and MATLAB (R) routines. The amplitude of bone oscillations (at twice the angular frequency of the rotating magnet) showed a characteristic pattern of resonance with the magnet rotational frequency. The amplitude maximum agreed with that calculated from a model of forced, damped harmonic oscillator and the diamagnetic tension force acting on bone. The calculated mean volume magnetic susceptibility (SI) of -was on the order of magnitude of literature value (vertical bar chi vertical bar approximate to 10(-5)). The results point to the bone site-specific aspect of LF PEMF therapy and provide experimental and theoretical basis for a novel, rational method of designing next generation of effective LF PEMF therapies.

The enumeration of circulating tumor cells (CTCs) in the peripheral bloodstream of metastatic cancer patients has contributed to improvements in prognosis and therapeutics. There have been numerous approaches to capture and counting of CTCs. However, CTCs have potential information beyond simple enumeration and hold promise as a liquid biopsy for cancer and a pathway for personalized cancer therapy by detecting the subset of CTCs having the highest metastatic potential. There is evidence that epithelial cell adhesion molecule (EpCAM) expression level distinguishes these highly metastatic CTCs. The few previous approaches to selective CTC capture according to EpCAM expression level are reviewed. A new two-stage microfluidic device for separation, enrichment and release of CTCs into subpopulations sorted by EpCAM expression level is presented here. It relies upon immunospecific magnetic nanoparticle labeling of CTCs followed by their field- and flow-based separation in the first stage and capture as discrete subpopulations in the second stage. To fine tune the separation, the magnetic field profile across the first stage microfluidic channel may be modified by bonding small Vanadium Permendur strips to its outer walls. Mathematical modeling of magnetic fields and fluid flows supports the soundness of the design.

The ability to separate RBCs from the other components of whole blood has a number of useful clinical and research applications ranging from removing RBCs from typical clinical blood draw, bone marrow transplants to transfusions of these RBCs to patients after significant blood loss. Viewed from a mechanistic/process perspective, there are three routine methodologies to remove RBCs: 1) RBCs lysis, 2) separation of the RBCs from the nucleated cells (i.e., stem cells) based on density differences typically facilitated through centrifugation or sedimentation agents, and 3) antibody based separation in which a targeted RBC is bound with an affinity ligand that facilitates its removal. More recently, several microfluidic based techniques have also been reported. In this report, we describe the performance of continuous RBC separation achieved by the deflection of intrinsically magnetic, deoxygenated RBCs as they flow through a magnetic energy gradient created by quadrupole magnet. This quadrupole magnetic, with aperture of 9.65 mm, has a maximum field of B0  = 1.36 T at the pole tips and a constant field gradient of B0 /r0  = 286 T/m. The annular flow channel, contained within this quadrupole magnet, is 203 mm long, has an inner radius of 3.98 mm, and an inner, outer radius of 4.36 mm, which corresponds to an annulus radius of 380 micrometer. At the entrance and exit to this annular channel, a manifold was designed which allows a cell suspension and sheath fluid to be injected, and a RBC enriched exit flow (containing the magnetically deflected RBCs) and a RBC depleted exit flow to be collected. Guided by theoretical models previously published, a limited number of operating parameters; total flow rate, flow rate ratios of flows in and flow out, and ratios of RBC to polystyrene control beads was tested. The overall performance of this system is consistent with our previously presented, theoretical models and our intuition. As expected, the normalized recovery of RBCs in the RBC exit fraction ranged from approximately 95% down to 60%, as the total flow rate through the system increased from 0.1 to 0.6 ml/min. At the cell concentrations studied, this corresponds to a flow rate of 1.5 × 106 -9 × 106  cells/min. While the throughput of these pilot scale studies are slow for practical applications, the general agreement with theory, and the small cross-sectional area in which the actual separation is achieved, 77 mm2 (annulus radius times the length), and corresponding volume of approximately 2 mls, suggests the potential to scale-up a system for practical applications exists and is actively being pursued.

Magnetic separation of cells has been, and continues to be, widely used in a variety of applications, ranging from healthcare diagnostics to detection of food contamination. Typically, these technologies require cells labeled with antibody magnetic particle conjugate and a high magnetic energy gradient created in the flow containing the labeled cells (i.e., a column packed with magnetically inducible material), or dense packing of magnetic particles next to the flow cell. Such designs, while creating high magnetic energy gradients, are not amenable to easy, highly detailed, mathematic characterization. Our laboratories have been characterizing and developing analysis and separation technology that can be used on intrinsically magnetic cells or spores which are typically orders of magnitude weaker than typically immunomagnetically labeled cells. One such separation system is magnetic deposition microscopy (MDM) which not only separates cells, but deposits them in specific locations on slides for further microscopic analysis. In this study, the MDM system has been further characterized, using finite element and computational fluid mechanics software, and separation performance predicted, using a model which combines: 1) the distribution of the intrinsic magnetophoretic mobility of the cells (spores); 2) the fluid flow within the separation device; and 3) accurate maps of the values of the magnetic field (max 2.27 T), and magnetic energy gradient (max of 4.41 T2 /mm) within the system. Guided by this model, experimental studies indicated that greater than 95% of the intrinsically magnetic Bacillus spores can be separated with the MDM system. Further, this model allows analysis of cell trajectories which can assist in the design of higher throughput systems.

Paramagnetic constituents of a cell have strong effect on cell's volume magnetic susceptibility even at low volume fraction because of their high susceptibility relative to that of the diamagnetic cell constituents. The effect can be measured at a single cell level by measuring cell terminal velocity in viscous media using a microscope equipped with a well-defined field and gradient magnet configuration (referred to as magnetophoretic analysis by cell tracking velocimetry, CTV). The sensitivity of such a microscopic-scale magnetometry was compared to that of a reference method of superconducting quantum interference-magnetic properties measurement system (SQUID-MPMS) using a red blood cell (RBC) suspension model. The RBC hemoglobin oxygen saturation determines the hemoglobin molecular magnetic susceptibility (diamagnetic when fully oxygenated, paramagnetic when fully deoxygenated or converted to methemoglobin). The SQUID-MPMS measurements were performed on an average of 5,000 RBCs in 20 μL physiological phosphate buffer at room temperature, those by CTV on a single cell track in a mean magnetic field of 1.6 T and mean gradient of 240 T/m, repeated for an average of 1,000 tracks per sample. This suggests 5,000× higher sensitivity of cell susceptometry by magnetophoretic analysis than by SQUID-MPMS. The magnetophoretic mean RBC magnetic susceptibilities were in the range determined by SQUID-MPMS (lower limit) and theory (upper limit). The ability of magnetophoretic analysis to resolve susceptibility peaks in a mixed cell populations was confirmed for an oxy RBC and met RBC mixture. Magnetophoretic analysis by CTV provides new tool for studies of emergence of paramagnetic reaction products in the cell.

The magnetic characteristics of hemoglobin (Hb) changes with the binding of dioxygen (O-2) to the heme prosthetic groups of the globin chains: from paramagnetic ferrous Hb to diamagnetic ferrous oxyhemoglobin (oxyHb) with reversibly bound O-2, or paramagnetic ferric methemoglobin (metHb). When multiplied over the number of Hb molecules in a red blood cell (RBC), the effect is detectable through motion analysis of RBCs in a high magnetic field and gradient. This motion is referred to as magnetophoretic mobility, which can be conveniently expressed as a fraction of the cell sedimentation velocity. In this Article, using a previously developed and reported instrument, cell tracking velocimetry (CTV), we are able to detect difference in Hb concentration in two RBC populations to a resolution of 1 X 10(7) Hb molecules per cell (4 X 10(7) atoms of Fe per cell or 4-5 femtograms of Fe). Similar resolution achieved with inductively coupled plasma-mass spectrometry requires on the order of 10(5)-10(6) cells and provides an average, whereas CTV provides a measurement for each cell. CTV analysis revealed that RBCs lose, on average, 17% of their Hb after 42 days of storage, the maximum FDA-approved length of time for the cold storage of RBCs in additive solution. This difference in Hb concentration was the result of routine RBC storage; clinical implications are discussed.

Arresting or regressing growth of abdominal aortic aneurysms (AAAs), localized expansions of the abdominal aorta are contingent on inhibiting chronically overexpressed matrix metalloproteases (MMPs)-2 and-9 that disrupt elastic matrix within the aortic wall, concurrent with providing a stimulus to augmenting inherently poor auto-regeneration of these matrix structures. In a recent study we demonstrated that localized, controlled and sustained delivery of doxycycline (DOX; a tetracycline-based antibiotic) from poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs), enhances elastic matrix deposition and MMP-inhibition at a fraction of the therapeutically effective oral dose. The surface functionalization of these NPs with cationic amphiphiles, which enhances their arterial uptake, was also shown to have pro-matrix regenerative and anti-MMP effects independent of the DOX. Based on the hypothesis that the incorporation of superparamagnetic iron oxide NPs (SPIONs) within these PLGA NPs would enhance their targetability to the AAA site under an applied external magnetic field, we sought to evaluate the functional effects of NPs co-encapsulating DOX and SPIONs (DOX-SPION NPs) on elastic matrix regeneration and MMP synthesis/activity in vitro within aneurysmal smooth muscle cell (EaRASMC) cultures. The DOX-SPION NPs were mobile under an applied external magnetic field, while enhancing elastic matrix deposition 1.5-2-fold and significantly inhibiting MMP-2 synthesis and MMP-2 and-9 activities, compared to NP-untreated control cultures. These results illustrate that the multifunctional benefits of NPs are maintained following SPION co-incorporation. Additionally, preliminary studies carried out demonstrated enhanced targetability of SPION-loaded NPs within proteolytically-disrupted porcine carotid arteries ex vivo, under the influence of an applied external magnetic field. Thus, this dual-agent loaded NP system proffers a potential non-surgical option for treating small growing AAAs, via controlled and sustained drug release from multifunctional, targetable nanocarriers.Statement of SignificanceProactive screening of high risk elderly patients now enables early detection of abdominal aortic aneurysms (AAAs). There are no established drug-based therapeutic alternatives to surgery for AAAs, which is unsuitable for many elderly patients, and none which can achieve restore disrupted and lost elastic matrix in the AAA wall, which is essential to achieve growth arrest or regression. We have developed a first generation design of polymer nanoparticles (NPs) for AAA tissue localized delivery of doxycycline, a modified tetracycline drug at low micromolar doses at which it provides both pro-elastogenic and antiproteolytic benefits that can augment elastic matrix regenerative repair. The nanocarriers themselves are also uniquely chemically functionalized on their surface to also provide them pro-elastin-regenerative & anti-matrix degradative properties. To provide an active driving force for efficient uptake of intralumenally infused NPs to the AAA wall, in this work, we have rendered our polymer NPs mobile in an applied magnetic field via co-incorporation of super-paramagnetic iron oxide NPs. We demonstrate that such modifications significantly improve wall uptake of the NPs with no significant changes to their physical properties and regenerative benefits. Such NPs can potentially stimulate structural repair in the AAA wall following one time infusion to delay or prevent AAA growth to rupture. The therapy can provide a non-surgical treatment option for high risk AAA patients. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

BACKGROUND:Conventional malaria parasite detection methods, such as rapid diagnostic tests (RDT) and light microscopy (LM), are not sensitive enough to detect low level parasites and identification of gametocytes in the peripheral blood. A modified and sensitive laboratory prototype, Magnetic Deposition Microscopy (MDM) was developed to increase the detection of sub-microscopic parasitaemia and estimation of gametocytes density in asymptomatic school children.METHODS:Blood samples were collected from 303 asymptomatic school children from seven villages in Bagamoyo district in Tanzania. Participants were screened for presence of malaria parasites in the field using RDT and MDM whereas further examination of malaria parasites was done in the laboratory by LM. LM and MDM readings were used to calculate densities and estimate prevalence of asexual and sexual stages of the parasite.RESULTS:Plasmodium falciparum parasites (asexual and sexual stages) were detected in 23 (7.6 %), 52 (17.2 %), and 59 (19.5 %) out of 303 samples by LM, RDT and MDM respectively. Gametocytes were detected in 4 (1.3 %) and 12 (4.0 %) out of the same numbers of samples by LM, and MDM, respectively. Likewise, in vitro results conducted on two laboratory strains of P. falciparum, 3D7 and NF54 to assess MDM sensitivity on gametocytes detection and its application on concentrating gametocytes indicated that gametocytes were enriched by MDM by 10-fold higher than LM. Late stages of the parasite strains, 3D7 and NF54 were enriched by MDM by a factor of 20.5 and 35.6, respectively. MDM was more specific than LM and RDT by 87.5 % (95 %, CI 71.2-89.6 %) and 89.0 % (95 % CI 82.9-91.4) respectively. It was also found that MDM sensitivity was 62.5 % (95 % CI 49.5-71.8) when compared with RDT while with LM was 36.5 % (95 % CI 32.2-60.5).CONCLUSIONS:These findings provide strong evidence that MDM enhanced detection of sub-microscopic P. falciparum infections and estimation of gametocyte density compared to current malaria diagnostic tools. In addition, MDM is superior to LM in detecting sub-microscopic gametocytaemia. Therefore, MDM is a potential tool for low-level parasitaemia identification and quantification with possible application in malaria transmission research.

Emerging microfluidic-based cell assays favor label-free red blood cell (RBC) depletion. Magnetic separation of RBC is possible because of the paramagnetism of deoxygenated hemoglobin but the process is slow for open-gradient field configurations. In order to increase the throughput, periodic arrangements of the unit magnets were considered, consisting of commercially available Nd-Fe-B permanent magnets and soft steel flux return pieces. The magnet design is uniquely suitable for multiplexing by magnet tessellation, here meaning the tiling of the magnet assembly cross-sectional plane by periodic repetition of the magnet and the flow channel shapes. The periodic pattern of magnet magnetizations allows a reduction of the magnetic material per channel with minimal distortion of the field cylindrical symmetry inside the magnet apertures. A number of such magnet patterns are investigated for separator performance, size and economy with the goal of designing an open-gradient magnetic separator capable of reducing the RBC number concentration a hundred-fold in 1mL whole blood per hour.

Abdominal aortic aneurysms (AAA) represent abnormal aortal expansions that result from chronic proteolytic breakdown of elastin and collagen fibers by matrix metalloproteases. Poor elastogenesis by adult vascular smooth muscle cells (SMCs) limits regenerative repair of elastic fibers, critical for AAA growth arrest. Toward overcoming these limitations, we recently demonstrated significant elastogenesis by bone marrow mesenchymal stem cell-derived SMCs (BM-SMCs) and their proelastogenesis and antiproteolytic effects on rat aneurysmal SMCs (EaRASMCs). We currently investigate the effects of super paramagnetic iron oxide nanoparticle (SPION) labeling of BM-SMCs, necessary to magnetically guide them to the AAA wall, on their functional benefits. Our results indicate that SPION-labeling is noncytotoxic and does not adversely impact the phenotype and elastogenesis by BM-SMCs. In addition, SPION-BM-SMCs showed no changes in the ability of the BM-SMCs to stimulate elastin regeneration and attenuate proteolytic activity by EaRASMCs. Together, our results are promising toward the utility of SPIONs for magnetic targeting of BM-SMCs for in situ AAA regenerative repair.

Low-frequency (LF), low-amplitude alternating current (ac) magnetic fields are approved by the U.S. Food and Drug Administration as an adjunct to the orthopedic surgery procedure (spinal fusion surgery) and for the treatment of selected bone healing complications (fracture non-union). The biological effects are understood to rely on a contact-less induction of electric currents in the electrically conducting compartments of the bone. Here, we investigate another potential mechanism of the LF magnetic field effect on bone by measuring mechanical force on bone due to its diamagnetic susceptibility. The biological rationale is the bone's exquisite sensitivity to local mechanical stresses that determine its structure and physiology. Thus, the periodic magnetic microstress effects on bone over an extended period of the typical clinical LF magnetic field application (hours daily over a period of a few months) could contribute to bone regeneration. The aim was to demonstrate the diamagnetic field effect on thin, human cortical bone slice preparations (from commercial tissue repository) suspended in the vicinity of a rotating cylindrical, diametrically magnetized permanent magnet. The data provide a basis for the calculation of diamagnetic microstresses generated by the clinical LF magnetic field stimulation devices and their significance as potential contributing factors to the biological mechanism of action of LF ac magnetic fields.

Tracking stem cells in vivo using non-invasive techniques is critical to evaluate the efficacy and safety of stem cell therapies. Superparamagnetic iron oxide nanoparticles (SPIONs) enable cells to be tracked using magnetic resonance imaging (MRI), but to obtain detectable signal cells need to be labelled with a sufficient amount of iron oxide. For the majority of SPIONs, this can only be obtained with the use of transfection agents, which can adversely affect cell health. Here, we have synthesised a library of dextran-based polymer coated SPIONs with varying surface charge from -1.5 mV to +18.2 mV via a co-precipitation approach and investigated their ability to be directly internalised by stem cells without the need for transfection agents. The SPIONs were colloidally stable in physiological solutions. The crystalline phase of the particles was confirmed with powder X-ray diffraction and their magnetic properties were characterised using SQUID magnetometry and magnetic resonance. Increased surface charge led to six-fold increase in uptake of particles into stem cells and higher MRI contrast, with negligible change in cell viability. Cell tracking velocimetry was shown to be a more accurate method for predicting MRI contrast of stem cells compared to measuring iron oxide uptake through conventional bulk iron quantification.

Algae were investigated in the past as a potential source of biofuel and other useful chemical derivatives. Magnetic separation of algae by iron oxide nanoparticle binding to cells has been proposed by others for dewatering of cellular mass prior to lipid extraction. We have investigated feasibility of magnetic separation based on the presence of natural iron stores in the cell, such as the ferritin in Auxenochlorella protothecoides (A. p.) strains. The A. p. cell constructs were tested for inserted genes and for increased intracellular iron concentration by inductively coupled plasma atomic absorption (ICP-AA). They were grown in Sueoka's modified high salt media with added vitamin B1 and increasing concentration of soluble iron compound (FeCl3 EDTA, from 1× to 8× compared to baseline). The cell magnetic separation conditions were tested using a thin rectangular flow channel pressed against interpolar gaps of a permanent magnet forming a separation system of a well-defined fluid flow and magnetic fringing field geometry (up to 2.2 T and 1,000 T/m) dubbed "magnetic deposition microscopy", or MDM. The presence of magnetic cells in suspension was detected by formation of characteristic deposition bands at the edges of the magnet interpolar gaps, amenable to optical scanning and microscopic examination. The results demonstrated increasing cellular Fe uptake with increasing Fe concentration in the culture media in wild type strain and in selected genetically-modified constructs, leading to magnetic separation without magnetic particle binding. The throughput in this study is not sufficient for an economical scale harvest.