Effectiveness of magnetically aligned collagen for neural regeneration in vitro and in vivo.

We investigated the effectiveness of using magnetically aligned collagen (after exposure to a maximum 8-T magnetic field) for nerve regeneration in both an in vitro and in vivo model. Neurite outgrowth from embryonic chick dorsal root ganglion (DRG) neurons was significantly greater on magnetically aligned collagen gel than on control gel, and was dependent on magnetic field strength. Silicone tubes (15 mm length) filled with collagen gel formed bridges between severed rat sciatic nerves. We prepared tubes for four groups: collagen gel only (COL), magnetically aligned collagen gel (M-COL), collagen gel mixed with Schwann cells (S-COL), and magnetically aligned collagen gel mixed with Schwann cells (M-S-COL). The ratio of infiltrating regenerated nerves was higher in the M-COL group compared to the COL group at 8 weeks post-operation. There were no significant differences between the two groups with and without Schwann cells. Compound action potentials showed higher amplitude and shorter latency in the M-COL than COL group at 12 weeks post-operation. The number and diameter of regenerated axons increased significantly in the M-COL compared with the COL group at 12 weeks post-operation. Here we demonstrated that magnetically orientated collagen promoted nerve regeneration using both an in vitro and in vivo model.

Impact of Titanium Skull Plate on Transcranial Magnetic Stimulation: Analysis of Induced Electric Fields.

BACKGROUND: Implanted titanium skull plates (TSPs) in cranioplasty are used to replace or reconstruct areas of the skull that have been damaged or removed due to trauma, surgery, or other medical conditions. However, the presence of a TSP in the head may influence the distribution of the electric field induced during transcranial magnetic stimulation (TMS) procedures. The purpose of this study was to determine how the presence of TSP would interfere with TMS-induced cortical electric fields. METHODS: The TMS with a figure-of-eight coil was applied to a realistic head model with TSPs. The distribution of the induced electric field in head tissues was calculated by employing the impedance method, and the results were compared with that of a normal head without TSP. RESULTS: Simulation results show that the distribution of the induced electric field has changed greatly for the head model with TSP. The maximum value of the induced electric field in head tissues was present under one of the circular coil wings rather than in the tissues beneath the junction of the two wings of the Fo8 coil. CONCLUSIONS: The induced electric field in deep brain regions was increased for the head model with TSP, which could potentially lead to deep brain stimulation. Since the presence of metallic TSP can greatly influence the distribution of the induced electric field in TMS applications, it is important to adjust the treatment scheme when considering TMS for individuals with cranial titanium plates.

Studies on magnetism and bioelectromagnetics for 45 years: from magnetic analog memory to human brain stimulation and imaging.

Forty-five years of studies on magnetism and bioelectromagnetics, in our laboratory, are presented. This article is prepared for the d'Arsonval Award Lecture. After a short introduction of our early work on magnetic analog memory, we review and discuss the following topics: (1) Magnetic nerve stimulation and localized transcranial magnetic stimulation (TMS) of the human brain by figure-eight coils; (2) Measurements of weak magnetic fields generated from the brain by superconducting quantum interference device (SQUID) systems, called magnetoencephalography (MEG), and its application in functional brain studies; (3) New methods of magnetic resonance imaging (MRI) for the imaging of impedance of the brain, called impedance MRI, and the imaging of neuronal current activities in the brain, called current MRI; (4) Cancer therapy and other medical treatments by pulsed magnetic fields; (5) Effects of static magnetic fields and magnetic control of cell orientation and cell growth; and (6) Effects of radio frequency magnetic fields and control of iron ion release and uptake from and into ferritins, iron cage proteins. These bioelectromagnetic studies have opened new horizons in magnetism and medicine, in particular for brain research and treatment of ailments such as depression, Parkinson's, and Alzheimer's diseases.

Figure-Eight Coils for Magnetic Stimulation: From Focal Stimulation to Deep Stimulation.

This article reviews the evolution and recent developments of transcranial magnetic brain stimulation using figure-eight coils to stimulate localized areas in the human brain. Geometric variations of figure-eight coils and their characteristics are reviewed and discussed for applications in neuroscience and medicine. Recent topics of figure-eight coils, such as focality of figure-eight coils, tradeoff between depth and focality, and approaches for extending depth, are discussed.

Synaptic Effect of Aδ-Fibers by Pulse-Train Electrical Stimulation.

Electrical stimulation of specific small fibers (Aδ- and C-fibers) is used in basic studies on nociception and neuropathic pain and to diagnose neuropathies. For selective stimulation of small fibers, the optimal stimulation waveform parameters are an important aspect together with the study of electrode design. However, determining an optimal stimulation condition is challenging, as it requires the characterization of the response of the small fibers to electrical stimulation. The perception thresholds are generally characterized using single-pulse stimulation based on the strength-duration curve. However, this does not account for the temporal effects of the different waveforms used in practical applications. In this study, we designed an experiment to characterize the effects of multiple pulse stimulation and proposed a computational model that considers electrostimulation of fibers and synaptic effects in a multiscale model. The measurements of perception thresholds showed that the pulse dependency of the threshold was an exponential decay with a maximum reduction of 55%. In addition, the frequency dependence of the threshold showed a U-shaped response with a reduction of 25% at 30 Hz. Moreover, the computational model explained the synaptic effects, which were also confirmed by evoked potential recordings. This study further characterized the activation of small fibers and clarified the synaptic effects, demonstrating the importance of waveform selection.

Dosimetry Analysis in Non-brain Tissues During TMS Exposure of Broca's and M1 Areas.

For human protection, the internal electric field is used as a dosimetric quantity for electromagnetic fields lower than 5-10 MHz. According to international standards, in this frequency range, electrostimulation is the main adverse effect against which protection is needed. One of the topics to be investigated is the quantification of the internal electric field threshold levels of perception and pain. Pain has been reported as a side effect during transcranial magnetic stimulation (TMS), especially during stimulation of the Broca's (speech) area of the brain. In this study, we designed an experiment to conduct a dosimetry analysis to quantify the internal electric field corresponding to perception and pain thresholds when targeting the Broca's and M1 areas from magnetic stimulator exposure. Dosimetry analysis was conducted using a multi-scale analysis in an individualized head model to investigate electrostimulation in an axonal model. The main finding is that the stimulation on the primary motor cortex has higher perception and pain thresholds when compared to Broca's area. Also, TMS-induced electric field applied to Broca's area exhibited dependence on the coil orientation at lower electric field threshold which was found to be related to the location and thickness of pain fibers. The derived dosimetry quantities provide a scientific rationale for the development of human protection guidelines and the estimation of possible side effects of magnetic stimulation in clinical applications.

Radio frequency magnetic field effects on molecular dynamics and iron uptake in cage proteins.

The protein ferritin has a natural ferrihydrite nanoparticle that is superparamagnetic at room temperature. For native horse spleen ferritin, we measure the low field magnetic susceptibility of the nanoparticle as 2.2 x 10(-6) m(3) kg(-1) and its Néel relaxation time at about 10(-10) s. Superparamagnetic nanoparticles increase their internal energy when exposed to radio frequency magnetic fields due to the lag between magnetization and applied field. The energy is dissipated to the surrounding peptidic cage, altering the molecular dynamics and functioning of the protein. This leads to an increased population of low energy vibrational states under a magnetic field of 30 microT at 1 MHz, as measured via Raman spectroscopy. After 2 h of exposure, the proteins have a reduced iron intake rate of about 20%. Our results open a new path for the study of non-thermal bioeffects of radio frequency magnetic fields at the molecular scale.

Short-term exposure to a 1439-MHz TDMA signal exerts no estrogenic effect in rats.

The aim of this study was to elucidate the possible effects of short-term exposure to a 1439-MHz electromagnetic field (EMF) employing time division multiple access (TDMA), which is the basis of the Japanese Personal Digital Cellular system, on estrogenic activity in rats. Sixty-four ovariectomized female Sprague-Dawley rats were divided into four groups: EMF exposure (EM), sham exposure, cage control, and 17 beta-estradiol injected (E2). The EM group was exposed, for 4 h per day on three consecutive days, to the 1439-MHz TDMA signal that produced 5.5-6.1 and 0.88-0.99 W/kg average specific absorption rates in the brain and the whole body, respectively. The uterine wet mass and serum estradiol level significantly increased in the E2 group, while there were no differences among the other three groups. Although negative effects of long-term EMF exposure must be thoroughly investigated before a final conclusion can be reached, our results do not support the assumption that the high frequency EMF used in cellular phones exerts estrogenic activity.

Group-level analysis of induced electric field in deep brain regions by different TMS coils.

Deep transcranial magnetic stimulation (dTMS) is a non-invasive technique used for the treatment of depression and obsessive compulsive disorder. In this study, we computationally evaluated group-level dosage for dTMS to characterize the targeted deep brain regions to overcome the limitations of using individualized head models to characterize coil performance in a population. We used an inter-subject registration method adapted to the deep brain regions that enable projection of computed electric fields (EFs) from individual realistic head models (n = 18) to the average space of deep brain regions. The computational results showed consistent group-level hotspots of the EF in the deep brain regions. The halo circular assembly coils induced the highest EFs in deep brain regions (up to 50% of the maximum EF in the cortex) for optimized positioning. In terms of the trade-off between field spread and penetration, the performance of the H7 coil was the best. The computational model allowed the optimization of generalized dTMS-induced EF on deep region targets despite inter-individual differences while informing and possibly minimizing unintended stimulation of superficial regions and possible mixed stimulation effects from deep and cortical areas. These results will facilitate the decision process during dTMS interventions in clinical practice.

Electrical Characterisation of Aδ-Fibres Based on Human in vivo Electrostimulation Threshold.

Electrical stimulation of small fibres is gaining attention in the diagnosis of peripheral neuropathies, such as diabetes mellitus, and pain research. However, it is still challenging to characterise the electrical characteristics of axons in small fibres (Aδ and C fibres). In particular, in vitro measurement for human Aδ-fibre is difficult due to the presence of myelin and ethical reason. In this study, we investigate the in vivo electrical characteristics of the human Aδ-fibre to derive strength-duration (S-D) curves from the measurement. The Aδ-fibres are stimulated using coaxial planar electrodes with intraepidermal needle tip. For human volunteer experiments, the S-D curve of Aδ-fibre is obtained in terms of injected electrical current. With the computational analysis, the standard deviation of the S-D curve is mostly attributed to the thickness of the stratum corneum and depth of the needle tip, in addition to the fibre thickness. Then, we derive electrical parameters of the axon in the Aδ-fibre based on a conventional fibre model. The parameters derived here would be important in exploring the optimal stimulation condition of Aδ-fibres.

Effects of radio frequency magnetic fields on iron release from cage proteins.

Ferritin, the iron cage protein, contains a superparamagnetic ferrihydrite nanoparticle formed from the oxidation and absorption of Fe(2+) ions. This nanoparticle increases its internal energy when exposed to alternating magnetic fields due to magnetization lag. The energy is then dissipated to the surrounding proteic cage, affecting its functioning. In this article we show that the rates of iron chelation with ferrozine, an optical marker, are reduced by up to a factor of 3 in proteins previously exposed to radio frequency magnetic fields of 1 MHz and 30 microT for several hours. The effect is non-thermal and depends on the frequency-amplitude product of the magnetic field.

Low-frequency conductivity tensor of rat brain tissues inferred from diffusion MRI.

Conductivity tensor maps of the rat brain were obtained using diffusion magnetic resonance imaging (MRI). Signal attenuations in the cortex and the corpus callosum were measured using the stimulated echo acquisition mode (STEAM) sequence with b factors up to 6000 s/mm(2). Our previously published method was improved to infer 3 x 3 conductivity tensor at the low-frequency limit. The conductivity tensor of the tissue was inferred from the fast component of the diffusion tensor and a fraction of the fast component. The mean conductivity (MC) of the cortex and the corpus callosum was 0.52 and 0.62 S/m, respectively. Diffusion-weighted images were obtained with b factors up to 4500 s/mm(2). Conductivity tensor images were calculated from the fast diffusion tensor images. Tissues with highly anisotropic cellular structures, such as the corpus callosum, the internal capsule, and the trigeminal nerve, exhibited high anisotropy in conductivity. The resulting values corresponded to conductivities at the low-frequency limit because our method assumed electric currents flowing only through extracellular fluid.

Estimation of cell membrane permeability and intracellular diffusion coefficient of human gray matter.

The signal intensity of diffusion-weighted imaging (DWI) is sensitive to the intra- and extracellular diffusion coefficient of water and cell membrane permeability. We applied a method we proposed in previous papers to estimate noninvasively the membrane permeability and intracellular diffusion coefficient of normal human brain (gray matter) in 3 normal volunteers. We theoretically compared predicted signals and experiment results using a 1.5-tesla magnetic resonance (MR) imaging system. We acquired images using an echo planar imaging (EPI) sequence, applying motion-probing gradient (MPG) pulses in 3 directions. We periodically performed numerical simulations for various combinations of membrane permeability and intracellular diffusion coefficients using the finite-difference method. By minimizing the difference between signals obtained experimentally and those from numerical simulation, we could estimate membrane permeability (76+/-9 mm2/s mum) and intracellular diffusion coefficient (1.0+/-0.0 mm2/s) for the human brain. The estimated membrane permeability was the criterion value for diagnosing disease in gray matter.

Late response evoked by cerebellar stimuli: effect of optokinetic stimulation.

We previously demonstrated that late electromyographic responses with a latency of 100 ms were evoked bilaterally in soleus muscles following transcranial magnetic stimulation over the left cerebellum. Efferent fibers from the left cerebellum modulate vestibulospinal tract influences on the extensor muscles of the left hindlimb. Here, we investigated whether the vestibulospinal tract mediates this late response. We activated the vestibulospinal tract by optokinetic stimulation. Our results show that the latency of the soleus electromyographic response is shortened by optokinetic stimulation, but the latency of the motor response evoked by the corticospinal tract is unchanged. These findings support our hypothesis that vestibulospinal tracts mediate late electromyographic responses, and allow the development of techniques to assess the human vestibulospinal system function.

Distinct neural correlates underlying two- and three-dimensional mental rotations using three-dimensional objects.

Two strategies-motor and visual-are considered to be used for performing mental rotation. The former involves the functions of the motor-related areas of the brain, whereas the latter does not. It is known that subjects' experiences influence strategy selection during the mental rotation of three-dimensional (3D) shapes. However, it remains questionable as to whether the attributes of 3D objects enhance the motor-related activities. In this regard, using functional magnetic resonance imaging, we compared the brain activities during two types of mental rotations-two-dimensional (2D) and 3D rotations-using 3D objects. 2D rotation using 3D objects requires rotation in a screen plain, whereas 3D rotation requires in-depth rotation. Only 3D rotation implicitly requires subjects to construct and manipulate 3D images with visualizations of the hidden parts; this plays an important role in visuo-motor tasks such as preshaping. As a result, a wide area of the right superior parietal lobule (SPL) was activated in relation to a 2D rotation angle. Conversely, a wide area of the right dorsal premotor cortex (PMd) was activated in relation to a 3D rotation angle. The right PMd activity is related to visualization of the hidden parts of visual stimuli, which is required only for 3D rotation. This implies that task difficulty enhanced by rotation dimensionality is a major factor related to the selection of motor strategy. In addition, it implies that the right SPL and the right PMd play important roles in rotation imagery without visualization and in constructing and manipulating 3D images, respectively.

Fusion and molecular aspects of liposomal nanocarriers incorporated with isoprenoids.

The present study was conducted to investigate whether typical isoprenyl compounds (TICs) can control liposomal fusion reactions through changes in the physical properties of membranes. The fusion capabilities of TIC-incorporated liposomes were characterized by measuring the 13C spin-lattice relaxation times (13CT1) and the gel permeation chromatogram (GPC) patterns. The 13CT1 relaxivities of some of these TIC-liposomes were remarkably enhanced at 27 degrees C. The highest 13CT1 value obtained was for the beta-carotene-liposome, which ruptured, and was attributed to the highest membrane fusion reactivity. The other TIC-liposomes incorporated with alpha-tocopherol, canthaxanthin, or coenzyme Q10 also induced significant fusion and did not rupture in comparison with the beta-carotene-liposome. These results show that the incorporations of TICs into lipid bilayers are useful to control liposomal nanocarriers for suitable membrane packing and advantageous phase separation, which could affect membrane-related processes.

Iron hydroxide nanoparticles coated with poly(ethylene glycol)-poly(aspartic acid) block copolymer as novel magnetic resonance contrast agents for in vivo cancer imaging.

PEG-coated beta-FeOOH nanoparticles were prepared through electrostatic complex formation of iron oxide nanoparticles with poly(ethylene glycol)-poly(aspartic acid) block copolymer [PEG-P(Asp)] in distilled water. By dynamic light scattering (DLS) measurement, the nanopaticle size was determined to be 70 nm with narrow distribution. The FT-IR and zeta potential experimental results proved that PEG-PAsp molecules bound to the surface of the iron oxide nanoparticles via the coordination between the carboxylic acid residues in the PAsp segment of the block copolymer and the surface Fe of the beta-FeOOH nanoparticles. The PEG-coated nanoparticles revealed excellent solubility and stability in aqueous solution as well as in physiological saline. In vivo MRI experiments on tumor-bearing mice demonstrated that the PEG-coated nanoparticles prepared by the current approach achieved an appreciable accumulation into solid tumor, suggesting their potential utility as tumor-selective MRI contrast agents.

Comparison of the induced fields using different coil configurations during deep transcranial magnetic stimulation.

Stimulation of deeper brain structures by transcranial magnetic stimulation (TMS) plays a role in the study of reward and motivation mechanisms, which may be beneficial in the treatment of several neurological and psychiatric disorders. However, electric field distributions induced in the brain by deep transcranial magnetic stimulation (dTMS) are still unknown. In this paper, the double cone coil, H-coil and Halo-circular assembly (HCA) coil which have been proposed for dTMS have been numerically designed. The distributions of magnetic flux density, induced electric field in an anatomically based realistic head model by applying the dTMS coils were numerically calculated by the impedance method. Results were compared with that of standard figure-of-eight (Fo8) coil. Simulation results show that double cone, H- and HCA coils have significantly deep field penetration compared to the conventional Fo8 coil, at the expense of induced higher and wider spread electrical fields in superficial cortical regions. Double cone and HCA coils have better ability to stimulate deep brain subregions compared to that of the H-coil. In the mean time, both double cone and HCA coils increase risk for optical nerve excitation. Our results suggest although the dTMS coils offer new tool with potential for both research and clinical applications for psychiatric and neurological disorders associated with dysfunctions of deep brain regions, the selection of the most suitable coil settings for a specific clinical application should be based on a balanced evaluation between stimulation depth and focality.

The state of the art of nanobioscience in Japan.

This paper reviews a part of the state of the art of nanobioscience in Japan. The importance of combination and integration of interdisciplinary principles is emphasized for the development of nanobioscience. Biomagnetics, biomechanics, nanomachining, self-replicating cell model, neuronal network, drug delivery system, and tissue engineering are discussed.

Evaluation of diffusional anisotropy and microscopic structure in skeletal muscles using magnetic resonance.

The pulsed-gradient spin-echo (PGSE) nuclear magnetic resonance (NMR) method is used for detecting the diffusion of water molecules in biological tissues. Because tissues generally have diffusional anisotropy, their diffusion properties are denoted by a tensor. In this study, we evaluated the diffusional anisotropy and microscopic structure in atrophied skeletal muscles using the PGSE NMR method. The left sciatic nerve was severed in twelve 9-week-old rats. Neurotomy caused neurogenic muscular atrophy at the left gastrocnemius. At 2, 4 and 8 weeks after neurotomy, magnetic resonance signals were selectively acquired from a 2 x 2 x 2 mm(3) voxel, which was located on the left gastrocnemius. The diffusion tensor, the mean diffusivity (MD) and the fractional anisotropy (FA) were calculated from the signals. A theoretical model of the diffusion in muscles was derived from Tanner's equation. The muscle fiber diameter was estimated by fitting the model to the measured signals. The measurements were also performed for normal rats as controls. No significant difference was found in the MD and the estimated intracellular diffusion coefficient between the control group and the denervated group. The denervated group had significantly higher FA compared with the control group (P<.05). The estimated muscle fiber diameter of the denervated group was significantly smaller than the estimated value of the control group (P<.05). These differences were found at 8 weeks after neurotomy. The proposed method is effective for evaluating changes in the microscopic structure of skeletal muscles.