Gil Navon

Assessment of glycosaminoglycan concentration in vivo by chemical exchange-dependent saturation transfer (gagCEST).

Glycosaminogycans (GAGs) are involved in numerous vital functions in the human body. Mapping the GAG concentration in vivo is desirable for the diagnosis and monitoring of a number of diseases such as osteoarthritis, which affects millions of individuals. GAG loss in cartilage is typically an initiating event in osteoarthritis. Another widespread pathology related to GAG is intervertebral disk degeneration. Currently existing techniques for GAG monitoring, such as delayed gadolinium-enhanced MRI contrast (dGEMRIC), T1ρ, and 23Na MRI, have some practical limitations. We show that by exploiting the exchangeable protons of GAG one may directly measure the localized GAG concentration in vivo with high sensitivity and therefore obtain a powerful diagnostic MRI method.

Neotendon formation induced by manipulation of the Smad8 signalling pathway in mesenchymal stem cells

Tissue regeneration requires the recruitment of adult stem cells and their differentiation into mature committed cells. In this study we describe what we believe to be a novel approach for tendon regeneration based on a specific signalling molecule, Smad8, which mediates the differentiation of mesenchymal stem cells (MSCs) into tendon-like cells. A biologically active Smad8 variant was transfected into an MSC line that coexpressed the osteogenic gene bone morphogenetic protein 2 (BMP2). The engineered cells demonstrated the morphological characteristics and gene expression profile of tendon cells both in vitro and in vivo. In addition, following implantation in an Achilles tendon partial defect, the engineered cells were capable of inducing tendon regeneration demonstrated by double quantum filtered MRI. The results indicate what we believe to be a novel mechanism in which Smad8 inhibits the osteogenic pathway in MSCs known to be induced by BMP2 while promoting tendon differentiation. These findings may have considerable importance for the therapeutic replacement of tendons or ligaments and for engineering other tissues in which BMP plays a pivotal developmental role.

A decoupled coil detector array for fast image acquisition in magnetic resonance imaging

A method for magnetic resonance imaging (MRI) is investigated here, whereby an object is put under a homogeneous magnetic field, and the image is obtained by applying inverse source procedures to the data collected in an array of coil detectors surrounding the object. The induced current in each coil due to the precession of the magnetic dipole in each voxel depends on the characteristics of both the magnetic dipole frequency and strength, together with its distance from the coil, the coil direction in space, and the electrical properties of the coils. By calculating the induced current signals over an array of coil detectors, a relationship is established between the set of signals and the structure of the body under investigation. The linear relation can then be represented in matrix notation, and inversion of this matrix will produce an image of the body. Important problems which must be considered in the proposed method are signal-to-noise ratio (SNR) and coupling between adjacent coils. Solutions to these problems will provide a new method for obtaining an instantaneous image by NMR, with no need for gradient switching for encoding. A general algorithm for decoupling of the coils is presented and fast sampling of the signal, instead of filtering, is used in order to reduce both noise and numerical roundoff errors at the same time. Sensitivity considerations are made with respect to the number of coils that is required and its connection with coil radius and SNR. A computer simulation demonstrates the feasibility of this new modality. Based on the solutions presented here for the problems involved in the use of a large number of coils for a simultaneous recording of the signal, an improved method of multicoil recording is suggested, whereby it is combined with the conventional zeugmatographic method with read and phase gradients, to result in a novel method of magnetic resonance imaging. In the combined method, there are no phase-encoding gradients. Only a single slice-selecting gradient, to be followed by a single read-gradient. Instead of phase-encoding gradients, use is made of an equivalent number of coils. The number of coils now is reduced significantly. This method suggests a single slice image taken within a single echo time, and where a 128 x 128 resolution is possible with only 128 coils. The applicability of the method is based on a successful decoupling procedure for the detectors (coils and cables) and the availability of highly accurate, high-gain, low-noise amplifiers with a broad dynamic range.

Selective and nonselective proton spin-lattice relaxation studies of enzyme-substrate interactions

Abstract The selective and nonselective longitudinal relaxation rates were measured for glycyl- l -tyrosine protons in the presence of Zn-carboxypeptidase A. Unlike the non-selective relaxation rates, the selective rate was found to be particularly sensitive to the presence of the enzyme. From the nonselective/selective ratio, the correlation times of the free and bound substrate were evaluated.

NMR relaxation studies of intracellular Na+ in red blood cells.

The state of intracellular Na+ in human and dog erythrocytes was characterized by 23Na-NMR using dysprosium complexes as shift reagents. Intracellular Na+ concentrations were determined using integration of the inner Na+ NMR signals and measurements of the intracellular volume using 59Co-NMR of extracellular Co(CN)3-6. T2 was found to be significantly shorter than T1, indicating some binding to macromolecules. While the longitudinal magnetization decay follows a single exponential, the transverse magnetization could be fitted with a double-exponential function. It was shown that neither the binding to the inner side of the membrane nor binding to hemoglobin contributes to the relaxation enhancement.

Molecular imaging of tumors and metastases using chemical exchange saturation transfer (CEST) MRI

The two glucose analogs 2-deoxy-D-glucose (2-DG) and 2-fluoro-2-deoxy-D-glucose (FDG) are preferentially taken up by cancer cells, undergo phosphorylation and accumulate in the cells. Owing to their exchangeable protons on their hydroxyl residues they exhibit significant chemical exchange saturation transfer (CEST) effect in MRI. Here we report CEST-MRI on mice bearing orthotopic mammary tumors injected with 2-DG or FDG. The tumor exhibited an enhanced CEST effect of up to 30% that persisted for over one hour. Thus 2-DG/FDG CEST MRI can replace PET/CT or PET/MRI for cancer research in laboratory animals, but also has the potential to be used in the clinic for the detection of tumors and metastases, distinguishing between malignant and benign tumors and monitoring tumor response to therapy as well as tumors metabolism noninvasively by using MRI, without the need for radio-labeled isotopes.

Ultrasound focusing using magnetic resonance acoustic radiation force imaging: application to ultrasound transcranial therapy.

PURPOSE Magnetic resonance guided ultrasonic therapy is a promising minimally invasive technology for constantly growing variety of clinical applications. Delivery of focused ultrasound (FUS) energy to the targeted point with optimal intensity is highly desired; however, due to tissue aberrations, optimal focal intensity is not always achieved. Especially in transcranial applications, the acoustic waves are shifted and distorted mainly by the skull. In order to verify that magnetic resonance acoustic radiation force imaging (MR-ARFI) can be used as a focusing tool in transcranial treatments, such an imaging was appliedin vivo on a porcine brain via ex vivo human skull. Then, by the use of MR-ARFI technique, an improved ultrasound focusing algorithm is proposed and demonstrated for both transcranial and none brain applications. METHODS MR-ARFI images were acquired on a GE 1.5 T scanner equipped with InSightec FUS systems ExAblate 2000 and ExAblate 4000. Imaging was performed with MR-ARFI sequences of line-scan spin-echo and single-shot gradient-echo echo-planar. The in-plane resolution of both acquisitions was 0.9×0.9mm2. The total acquisition time of MR-ARFI image was 31 s by the line-scan sequence and 1 s by the echo-planar sequence. An in vivo experiment was performed using FUS transducer, which is built out of 1024 ultrasound transmitting piezoelectric elements at 220 kHz frequency. The transducer was focused into the brain of a pig, which was wrapped in a human skull, in degassed water environment to resemble human treatments. The pig underwent a wide bilateral craniectomy to prevent a bone heating from the ultrasound beams. Two focusing experiments were performed in phantoms using 1 MHz and 710 kHz FUS transducers working with 208 and 225 elements, respectively. In the first experiment, aberration was added virtually to the apparatus by adding random phases to the phase map of the transducer. A simple focusing correction scheme was used, in which the corrected phase of a group of elements was chosen such that it maximizes the radiation force at the focal point. In the second experiment, aberrations made by a human skull were corrected using geometrical and phase based adjustments on segments of the transducer. RESULTS A maximum displacement of 10μm was obtained using 1.4 kW acoustic power on a live pigs head that its skull was removed and replaced by ex vivo human skull. Aberration correction using MR-ARFI resulted in near optimal focus, as the radiation force was similar to the nonaberration case. Transcranial, MR-ARFI based aberration correction performed better than CT based aberration correction, a technique that is currently used in brain FUS treatments. CONCLUSIONS In the present work, the authors show for the first time a result of MR-ARFI in a live brain throughex vivo human skull. They have demonstrated that aberration correction could be done using MR-ARFI by measuring the radiation force at the focal point. Aberration correction using MR-ARFI is a promising noninvasive technique for transcranial focusing, which may result in near optimal focus and more reliable and safer brain FUS treatments.

Assessment of glycosaminoglycan concentration changes in the intervertebral disc via chemical exchange saturation transfer.

In this study, it is shown that the chemical exchange saturation transfer (CEST) method for hydroxyl protons can be used to detect changes in glycosaminoglycan (GAG) concentration in the intervertebral disc. The method, termed gagCEST, was demonstrated ex vivo by correlating the CEST effect with the fixed charge density (FCD) of the nucleus pulposus (NP), as well as by correlating tissue CEST images with their corresponding 23Na images. Incubation of five NP samples with trypsin produced samples with varying GAG content (n = 19). A good correlation was found between the –OH CEST effect and FCD, as well as with the N‐acetyl signal amplitude. gagCEST images in vitro further illustrated the amount of detail obtainable from this contrast mechanism when compared with conventional imaging. The large concentration of GAG and the relatively long T1 of water in NP make the method sensitive, in particular, for the assessment of GAG depletion in this tissue. It is the loss of GAG in NP that indicates the early stage of disc degeneration. Copyright

Study of order and dynamic processes in tendon by NMR and MRI.

Tendons are composed of a parallel arrangement of densely packed collagen fibrils that results in unique biomechanical properties of strength and flexibility. In the present review we discuss several advanced magnetic resonance spectroscopy (MRS) and imaging (MRI) techniques that have allowed us to better understand the biophysical properties of tendons and ligaments. The methods include multiple quantum and T2 filtering combined with NMR and MRI techniques. It is shown in detail how these techniques can be used to extract a number of useful parameters: 1) the 1H‐1H and 1H‐2H dipolar interactions; 2) the proton exchange rates between water and collagen, and between water molecules; 3) the distribution of fibril orientations; and 4) the anisotropy of diffusion. It is shown that relaxation data as a function of angular dependence can be obtained in vivo using mobile NMR sensors. Finally, this article describes how double quantum filtered (DQF) MRI can be used to image and monitor the healing process in injured tendons. J. Magn. Reson. Imaging 2007.

Mapping the fiber orientation in articular cartilage at rest and under pressure studied by 2H double quantum filtered MRI.

The one‐dimensional 2H double quantum filtered (DQF) spectroscopic imaging technique was used to study the orientation of collagen fibers in articular cartilage. The method detects only water molecules in anisotropic environments, which in cartilage is caused by their interaction with the collagen fibers. A large quadrupolar splitting was observed in the calcified zone and a smaller splitting in the radial zone. In the transitional zone the splitting was not resolved and a small splitting was again detected in the superficial zone. From measurements performed at two orientations of the plug relative to the magnetic field it was deduced that in the calcified and radial zones the fibers are oriented perpendicular to the bone, bending at the transitional zone and flattening at the superficial zone. The effect of load applied to the cartilage–bone plug was monitored by the same technique. At low loads there is a small decrease in the quadrupolar splitting in the calcified zone, a marked decrease in the radial zone, and an increase of the splitting accompanied by a thickening of the superficial zone. Under high loads, while the thickening and the splitting of the superficial zone further increase, the splitting in the radial and calcified zones completely collapse. Pressure‐induced changes in the thickness of the surface zone indicate flattening of the collagen fibers near the surface. The marked collapse of the splitting near the bone at high pressures may result from crimping of the collagen fibers. Magn Reson Med 48:322–330, 2002.