Evaluation of collagen fiber maturation and ordering in regenerating tendons employing H‐1 double quantum filtered NMR spectroscopy

Ikoma, Kazuya; Kusaka, Yoshiaki; Takamiya, Hisatake; Eliav, Uzi; Seo, Yoshiteru

doi:10.1016/s0736-0266(02)00092-x

Cited by 12 publications

(18 citation statements)

References 23 publications

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“…Cell pellets of transfected HEK 293T or C3H10T1/2 cells (shock-frozen in liquid nitrogen) were lysed in 1% Nonidet P-40, 150 mM NaCl, 20 mM Tris (pH 7.5), 2 mM EDTA, 50 mM NaF, 1 mM Na2P2O7, and 1 mM phenylmethylsulfonyl fluoride supplemented with protease inhibitors (Protease Inhibitor Mixture Tablets; Roche Molecular Biochemicals). Vectors encoding the constitutively active HA-tagged TGFβ/BMP type I receptors ALK1-ALK6 in pCDNA3 were obtained from P. ten Dijke (Leiden University Medical Center, Leiden, The Netherlands) and C. Heldin (Uppsala University, Uppsala, Sweden) (45), and 1 vector encoding ALK7 was obtained from C. Ibanez (Karolinska Institute, Stockholm, Sweden) (46). Immunoblotting with appropriate antibodies was used to analyze cell extracts.…”

Section: Methodsmentioning

confidence: 99%

“…DQF MRI provides high-contrast signals, differentiating the tissues of tendon, bone, skin, and muscle. Tendons, which produce a weak signal in standard MRI sequences, are highlighted in the 1 H DQF image (45,46).…”

Section: Methodsmentioning

confidence: 99%

“…Both the surgically treated leg and the intact control leg were subjected to a micro-MRI analysis, and multi-slice-multi-echo (MSME) images were obtained. The same samples were further subjected to DQF MRI, in which only the tendons were analyzed (46,47).…”

Section: Methodsmentioning

confidence: 99%

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

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Pelled²,

Turgeman³

et al. 2006

Journal of Clinical Investigation

222

166

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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

Hoffmann

Pelled²,

Turgeman³

et al. 2006

Journal of Clinical Investigation

222

166

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“…The DQF signal for the 15‐year‐old specimen is seen to retain a sizeable intensity up to evolution times as long as 100 ms. This is significantly longer than the build‐up times usually measured in connective tissues (7, 23–26). To verify that the observed signals at long τ times arise exclusively due to the DQ coherence, and there is no leakage of the SQ coherence, we examined the off‐resonance DQ evolution behavior as a function of the t 1 time, as described by Shinar et al (23).…”

Section: Resultsmentioning

confidence: 73%

“…The DQF experiment exclusively detects NMR signals of nuclei that experience nonzero residual dipolar or quadrupolar interactions arising from the anisotropic motions of bound water molecules, while free water, which has vanishing dipolar and quadrupolar interactions, is not represented in the DQF spectra (7). This method has found wide application in connective tissues, such as cartilage, tendon, and components of the sciatic nerve (4–7, 21–26). The spectra are measured as a function of a DQF build‐up time τ (also referred to as the creation time).…”

mentioning

confidence: 99%

²H double quantum filtered (DQF) NMR spectroscopy of the nucleus pulposus tissues of the intervertebral disc

Perea

Cannella

Yang

et al. 2007

Magnetic Resonance in Med

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Deuterium (2 H) double-quantum filtered (DQF) NMR spectros-copy of nucleus pulposus (NP) tissues from human interverte-bral discs is reported. The DQF spectral intensities, DQ build-up rates, and DQF-detected rotating-frame spin-lattice relaxation times are sensitive to the degree of hydration of the NP tissue, and display a monotonous correlation with age between 15 and 80 years. The implications of this work are that the changes in water dynamics as detected via DQF NMR spectroscopy may be used as a probe of tissue degeneration in NP, particularly in the early stages of degeneration to which most standard NMR methods are not sensitive. Magn Reson Med 57:990-999, 2007. The challenges to the physician in treating lower back pain due to degenerative disc disease (DDD) begin with diagnosis. Physical examination and MRI (T 2 imaging) are typical methods employed to detect a degenerated disc. However, little correlation exists between lower back pain and the structural changes that are visualized using current MRI techniques, leaving the clinician with less than desirable noninvasive diagnostic capabilities (1). Furthermore, de-generation of the disk is related to aging, but the correlation is not trivial, and multiple other factors contribute to the degeneration processes (2). To understand more detail about the degenerative state of the disc, an analysis of the water structure and dynamics in the disc may prove enlightening. Each tissue in the body has different forms of water within the cellular and extracellular matrixes that can be broadly categorized as bound, intermediately bound, or free water. Little is documented regarding the detailed water structure and dynamics of normal or degenerated nucleus pulposus (NP) of the intervertebral disc. The three main molecular constituents of the nucleus are collagen, proteoglycan, and water. The proteoglycan (aggrecan) consists of sulfated glycosaminoglycan (GAG) side chains attached to a protein core. The total water present in the nucleus is dependent, in part, on the charge density of the sulfated GAGs, which change in structure and reduce in quantity with degeneration of the NP, leading to a reduction in water content (3). Since changes in disc water content are critical for understanding the pathogenesis of DDD, it is important to consider the properties of this common molecule in the changing disc environment. 1 H and 2 H NMR spectroscopy have been established as probes of the dynamics of "bound" waters in the various connective tissues (4-7). For NMR investigations of the intervertebral disc, proton MRI has been the preferred method in nearly all published works (8-17). In an effort to develop imaging methods for the diagnosis of DDD (14), numerous quantitative MRI studies were undertaken to identify statistical correlations of the proton T 1 and T 2 relaxation times of the water signal with disc degradation grade (8,13,17), age (10), diurnal fluctuations (9,12), water content (8), GAG content (8), anatomical position (17), and mechanical properties (15). In addi...

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

Navon

Eliav

Demco

et al. 2007

Magnetic Resonance Imaging

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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 T 2 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 1 H-1 H and 1 H-2 H 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. TENDONS ARE THE FIBROUS CONNECTIVE TISSUE that joins muscle to bone. For example, the Achilles tendon connects the heel to the muscles of the calf (Fig. 1). Ligaments are similar to tendons, except that they join one bone to another. Tendons and ligaments are unique in that they possess biomechanical properties of exceptional strength and flexibility due to the parallel arrangement of densely packed type-I collagen fibrils (see Fig. 2). In addition to collagen, these tissues contain a small amount of proteoglycans and cells called tenocytes. Injuries of the tendons and ligaments are very common issues in geriatric and sports medicine, so the adaptation of medical imaging techniques to these tissues is a subject of considerable interest. These important tissues are difficult to image by MRI due to their short and anisotropic T 2 resulting from the residual dipolar interactions of the water molecules. In this article we review advanced MR spectroscopy (MRS) and MRI techniques that can be applied to the tendons and ligaments. These methods have already allowed us to better understand the biophysical properties of tendons, and to suggest new directions for imaging. MEASUREMENT OF DIPOLAR INTERACTIONS IN TENDON BY DOUBLE QUANTUM FILTERED (DQF) NMR SPECTROSCOPYThe interaction of water molecules with collagen fibers induces an anisotropy in their motions (3,4). Thus, the intramolecular dipolar interaction does not average to zero, and a smaller (residual) dipolar splitting remains. While water molecules that interact directly with the highly ordered fibers may exhibit a large dipolar splitting, a fast exchange process with adjacent isotropically moving water molecules will reduce the splitting (Fig. 3). It follows that the resulting residual dipolar splitting depends on the ratio of bound to free water molecules, and therefore on the hydration level of the sample. Splitting of the water 1 H and 2 H NMR signals due to residual dipolar or quadrupolar interactions has been observed (4,5) in partially hy...

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Evaluation of collagen fiber maturation and ordering in regenerating tendons employing H‐1 double quantum filtered NMR spectroscopy

Cited by 12 publications

References 23 publications

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

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

²H double quantum filtered (DQF) NMR spectroscopy of the nucleus pulposus tissues of the intervertebral disc

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

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Evaluation of collagen fiber maturation and ordering in regenerating tendons employing H‐1 double quantum filtered NMR spectroscopy

Cited by 12 publications

References 23 publications

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

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

2H double quantum filtered (DQF) NMR spectroscopy of the nucleus pulposus tissues of the intervertebral disc

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

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²H double quantum filtered (DQF) NMR spectroscopy of the nucleus pulposus tissues of the intervertebral disc