Nuclear Magnetic Resonance Microimaging Research Articles

Nuclear Magnetic Resonance Microimaging for the Qualitative Assessment of Root Canal Treatment: An Ex Vivo Preliminary Study.

Aim: To assess the potential ability of nuclear magnetic resonance micro-imaging (mMRI) to visualize and identify soft tissue debris and unfilled spaces inside radicular canals in endodontic treated extracted teeth, for understanding the causes of treatment failure. Toward this goal, multi-parametric mMRI and cone beam computed tomography (CBCT) were compared. Methodology: A non-recoverable root treated human tooth was extracted due to endodontic failure and excessive mobility. It was examined with both CBCT and mMRI: CBCT was performed with 0.125 mm voxel size (GXCB-500, Kavo-Gendex, Brea, CA, USA) and mMRI was performed with a spectrometer operating at 9.4T magnetic field (Bruker Avance-400, Bruker, Billerica, MA, USA). The mMRI images were obtained with a microimaging probe. Relaxation times (T1 and T2) and diffusion-weighted acquisition sequences were used to obtain multi-parametric maps of the extracted tooth (slice thickness of 200 µm and in plane resolution of 30 × 30 µm2). Results: T1 and T2 maps identified unfilled spaces around and close to Gutta-percha cones instead of CBCT images that were not able to highlight this aspect. T1, T2 and apparent diffusion coefficient (ADC) assumed different values in dentine and in voids, characterized by different dimensions. Moreover, they were able to discriminate between infiltrations of water only and deposits of biological material. Because Gutta-percha cones are constituted of hard, non-porous material, they do not provide a signal and in mMRI images appear as zones of noise. Conclusions: Unlike the CBCT exam, mMRI can detect soft tissue debris and unfilled spaces inside radicular canals. Therefore, this in vitro study showed the potential of mMRI to evaluate the quality of the root canal treatment, suggesting its potential benefit in determining the causes of endodontic failure, without the use of ionizing radiation.

Structural and metabolic responses of Staphylococcus aureus biofilms to hyperosmotic and antibiotic stress.

Biofilms alter their metabolism in response to environmental stress. This study explores the effect of a hyperosmotic agent-antibiotic treatment on the metabolism of Staphylococcus aureus biofilms through the use of nuclear magnetic resonance (NMR) techniques. To determine the metabolic activity of S. aureus, we quantified the concentrations of metabolites in spent medium using high-resolution NMR spectroscopy. Biofilm porosity, thickness, biovolume, and relative diffusion coefficient depth profiles were obtained using NMR microimaging. Dissolved oxygen concentration was measured to determine the availability of oxygen within the biofilm. Under vancomycin-only treatment, the biofilm communities switched to fermentation under anaerobic condition, as evidenced by high concentrations of formate (7.4 ± 2.7 mM), acetate (13.1 ± 0.9 mM), and lactate (3.0 ± 0.8 mM), and there was no detectable dissolved oxygen in the biofilm. In addition, we observed the highest consumption of pyruvate (0.19 mM remaining from an initial 40 mM concentration), the sole carbon source, under the vancomycin-only treatment. On the other hand, relative effective diffusion coefficients increased from 0.73 ± 0.08 to 0.88 ± 0.08 under vancomycin-only treatment but decreased from 0.71 ± 0.04 to 0.60 ± 0.07 under maltodextrin-only and from 0.73 ± 0.06 to 0.56 ± 0.08 under combined treatments. There was an increase in biovolume, from 2.5 ± 1 mm3 to 7 ± 1 mm3 , under the vancomycin-only treatment, while the maltodextrin-only and combined treatments showed no significant change in biovolume over time. This indicated that physical biofilm growth was halted during maltodextrin-only and combined treatments.

Modeling Substrate Utilization, Metabolite Production, and Uranium Immobilization in Shewanella oneidensis Biofilms

In this study, we developed a two-dimensional mathematical model to predict substrate utilization and metabolite production rates in Shewanella oneidensis MR-1 biofilm in the presence and absence of uranium (U). In our model, lactate and fumarate are used as the electron donor and the electron acceptor, respectively. The model includes the production of extracellular polymeric substances (EPS). The EPS bound to the cell surface and distributed in the biofilm were considered bound EPS (bEPS) and loosely associated EPS (laEPS), respectively. COMSOL® Multiphysics finite element analysis software was used to solve the model numerically (model file provided in the Supplementary Material). The input variables of the model were the lactate, fumarate, cell and EPS concentrations, half saturation constant for fumarate, and diffusion coefficients of the substrates and metabolites. To estimate unknown parameters and calibrate the model, we used a custom designed biofilm reactor placed inside a nuclear magnetic resonance (NMR) microimaging and spectroscopy system and measured substrate utilization and metabolite production rates. From these data we estimated the yield coefficients, maximum substrate utilization rate, half saturation constant for lactate, stoichiometric ratio of fumarate and acetate to lactate and stoichiometric ratio of succinate to fumarate. These parameters are critical to predicting the activity of biofilms and are not available in the literature. Lastly, the model was used to predict uranium immobilization in S. oneidensis MR-1 biofilms by considering reduction and adsorption processes in the cells and in the EPS. We found that the majority of immobilization was due to cells, and that EPS was less efficient at immobilizing U. Furthermore, most of the immobilization occurred within the top 10 μm of the biofilm. To the best of our knowledge, this research is one of the first biofilm immobilization mathematical models based on experimental observation. It has the ability to predict the relative contributions to U immobilization of laEPS, bEPS, and cells.

In situ nuclear magnetic resonance microimaging of live biofilms in a microchannel.

Biofilms are comprised of microbial cells and an extracellular polymeric substance (EPS) matrix that supports interactions between community members and with the local environment. The highly hydrated EPS matrix makes the application of many biofilm visualization techniques difficult. Hence, to better visualize how biofilms interact with their environment, there is a need for imaging techniques to monitor hydrated state biofilm dynamics. We employed an in situ dynamic approach to construct label-free images of biofilms. In situ imaging was conducted using a vacuum compatible microfluidic reactor, SALVI (System for Analysis at the Liquid Vacuum Interface), for biofilm growth; real-time confocal laser scanning microscopy analysis; and nuclear magnetic resonance (NMR) microimaging and spectroscopy. We integrated SALVI microchannel fluids and live biofilms to demonstrate in situ measurement capabilities, including velocity mapping, diffusion coefficient mapping, relaxometry, localized spectroscopy, relaxation times, porosity, and two- and three-dimensional imaging within the microchannel at high spatial resolution. We monitored organic acids adjacent to biofilms, suggesting that kinetic rate and substrate-product yield ratio studies are possible using the SALVI microfluidic reactor for growth characterizations. The integration of NMR microimaging studies into the SALVI platform demonstrates that a multimodal microfluidic platform can serve as an avenue to explore complex biological phenomena, such as biofilm attachment to surfaces, with detailed quantitative physical and chemical mapping. The further incorporation of other SALVI-compatible technologies, such as liquid time-of-flight secondary ion mass spectrometry imaging, with NMR microimaging will produce a powerful correlative approach to monitor in situ biofilm chemistry and dynamics at different spatial scales.

Free-evolution kinetics in a high-swelling polymeric hydrogel

We employ nuclear magnetic resonance microimaging to study the kinetics of a high-swelling ionic polymer gel. This includes the time evolution of the sphere diameter and also the evolution of the swollen-unswollen boundary. The experimental results for spherical ionic polyacrylamide gels are compared with the predictions of a nonlinear poroelastic theory by numerically solving the equations for the evolution.

Microvascular obstruction in the right ventricle in reperfused anterior myocardial infarction. Macroscopic and pathologic evidence in a swine model

Data on right ventricular (RV) involvement in anterior myocardial infarction are scarce. The presence of RV microvascular obstruction (MVO) in this context has not been analyzed yet. The aim of the present study was to characterize the presence of MVO in the RV in a controlled experimental swine model of reperfused anterior myocardial infarction. Left anterior descending (LAD) artery-perfused area (thioflavin-S staining after selective infusion in LAD artery), infarct size (lack of triphenyltetrazolium-chloride staining) and MVO (lack of thioflavin-S staining in the core of the infarcted area) in the RV were studied. A quantitative (% of the ventricular volume) and semiquantitative (number of segments involved) analysis was carried out both in the RV and LV in a 90-min left anterior descending balloon occlusion and 3-day reperfusion model in swine (n=15). RV infarction and RV MVO (>1 segment) were detected in 9 (60%) and 6 (40%) cases respectively. Mean LAD-perfused area, infarct size and MVO in the RV were 33.8 ± 13%, 13.53 ± 11.7% and 3.4 ± 4.5%. Haematoxylin and eosin stains and electron microscopy of the RV-MVO areas demonstrated generalized cardiomyocyte necrosis and inflammatory infiltration along with patched hemorrhagic areas. Ex-vivo nuclear magnetic resonance (T2 sequences) microimaging of RV-MVO showed, in comparison with remote non-infarcted territories, marked hypointense zones (corresponding to necrosis, inflammation and hemorrhage) in the core of hyperintense regions (corresponding to edema). In reperfused anterior myocardial infarction, MVO is frequently present in the RV. It is associated with severe histologic repercussion on the RV wall. Nuclear magnetic resonance appears as a promising technique for the noninvasive detection of this phenomenon. Further studies are warranted to evaluate the pathophysiological and clinical implications.

Effects of specific surface area and porosity on cube counting fractal dimension, lacunarity, configurational entropy, and permeability of model porous networks: Random packing simulations and NMR micro-imaging study

Despite the importance of understanding and quantifying the microstructure of porous networks in diverse geologic settings, the effects of the specific surface area and porosity on the key structural parameters of the networks have not been fully understood. We performed cube-counting fractal dimension (Dcc) and lacunarity analyses of 3D porous networks of model sands and configurational entropy analysis of 2D cross sections of model sands using random packing simulations and nuclear magnetic resonance (NMR) micro-imaging. We established relationships among porosity, specific surface area, structural parameters (Dcc and lacunarity), and the corresponding macroscopic properties (configurational entropy and permeability). The Dcc of the 3D porous networks increases with increasing specific surface area at a constant porosity and with increasing porosity at a constant specific surface area. Predictive relationships correlating Dcc, specific surface area, and porosity were also obtained. The lacunarity at the minimum box size decreases with increasing porosity, and that at the intermediate box size (∼0.469 mm in the current model sands) was reproduced well with specific surface area. The maximum configurational entropy increases with increasing porosity, and the entropy length of the pores decreases with increasing specific surface area and was used to calculate the average connectivity among the pores. The correlation among porosity, specific surface area, and permeability is consistent with the prediction from the Kozeny–Carman equation. From the relationship between the permeability and the Dcc of pores, the permeability can be expressed as a function of the Dcc of pores and porosity. The current methods and these newly identified correlations among structural parameters and properties provide improved insights into the nature of porous media and have useful geophysical and hydrological implications for elasticity and shear viscosity of complex composites of rock, glasses, melt, and fluids as well as transport behavior of ground water.

Diffusion in biofilms respiring on electrodes

The goal of this study was to measure spatially and temporally resolved effective diffusion coefficients (D(e)) in biofilms respiring on electrodes. Two model electrochemically active biofilms, Geobacter sulfurreducens PCA and Shewanella oneidensis MR-1, were investigated. A novel nuclear magnetic resonance microimaging perfusion probe capable of simultaneous electrochemical and pulsed-field gradient nuclear magnetic resonance (PFG-NMR) techniques was used. PFG-NMR allowed noninvasive, nondestructive, high spatial resolution in situ D(e) measurements in living biofilms respiring on electrodes. The electrodes were polarized so that they would act as the sole terminal electron acceptor for microbial metabolism. We present our results as both two-dimensional D(e) heat maps and surface-averaged relative effective diffusion coefficient (D(rs)) depth profiles. We found that 1) D(rs) decreases with depth in G. sulfurreducens biofilms, following a sigmoid shape; 2) D(rs) at a given location decreases with G. sulfurreducens biofilm age; 3) average D(e) and D(rs) profiles in G. sulfurreducens biofilms are lower than those in S. oneidensis biofilms-the G. sulfurreducens biofilms studied here were on average 10 times denser than the S. oneidensis biofilms; and 4) halting the respiration of a G. sulfurreducens biofilm decreases the D(e) values. Density, reflected by D(e), plays a major role in the extracellular electron transfer strategies of electrochemically active biofilms.

Surface pattern formation in a swelling gel

We study the time evolution of surface pattern formation in a swelling spherical gel using proton nuclear magnetic resonance microimaging of water molecules migrating into the gel. Relaxation-weighted images permit mapping of the local evolution of the swelling ratio which, within the framework of a nonlinear poroelastic theory, can be identified with the determinant of the deformation gradient tensor. The wavelength of the surface pattern is examined and some analogies with the extensively studied problem of the wavelength at threshold for swelling rings are pointed out. By controlling the degree of cross-linking within a surface layer of the gel it is possible to monitor the transition from an undulating folding pattern to the formation of singular creases.

Mathematical Models of Naturally “Morphed” Human Erythrocytes: Stomatocytes and Echinocytes

We present two mathematical models that describe human red blood cells (RBCs) with morphologies that are attained naturally under certain patho-physiological conditions, namely stomatocytes and echinocytes. Muñoz San Martín et al. (Bioelectromagnetics 27:521-527, 2006) recently presented models of these shapes based on our previous set of parametric equations (Kuchel and Fackerell, Bull. Math. Biol. 61:209-220, 1999) that involve Jacobi elliptic functions and integrals. Thus, both discocytes and stomatocytes are described. Here, we derived the Cartesian forms of these new equations; and, in addition, present a realistic model of a Type III echinocyte, using prolate spheroids 'decorating' a central sphere at the vertices of an internal dodecahedron. The RBC models based on Cartesian equations have been used for representing the shape changes (morphological transformations or "morphing") that occur in RBCs under various experimental conditions; specifically, when the shape changes have been monitored by nuclear magnetic resonance (NMR) micro-imaging.

Nuclear magnetic resonance microimaging investigation of membrane electrode assembly of fuel cells: morphology and solvent dynamics

The structure and solvent (water, methanol, etc.) dynamics of a number of fuel membrane electrode assembly (MEA) samples are studied with nuclear magnetic resonance microimaging with spatial resolution of tens of micrometers. The micrometer-scale inhomogeneity of the samples is observed and confirmed with various weighting methods. In particular, diffusion coefficients at different positions in MEA are clearly differentiated. Furthermore, chemical shift selection imaging enables one to investigate the spatial distribution and dynamics of individual chemical groups. These types of information offer us insights into the working principle of fuel cell and pave the way to in situ studies of operating fuel cells.

Phase separation in nematic microemulsions probed by one-dimensional spectroscopic deuteron magnetic resonance microimaging

We present a study of a phase-transition-driven separation in microemulsions of nanosized lyotropic inverse micelles and thermotropic liquid crystal pentylcyanobiphenyl (5CB) with 5%, 8%, and 15% micelle concentration. Using deuteron nuclear magnetic resonance (DNMR) microimaging in combination with conventional microscopy as well as ac calorimetry, we demonstrate a phase separation scenario in which micelles are expelled from the nematic phase during the I-N conversion. Due to a difference in density the micelle-rich isotropiclike phase spatially separates from the micelle-free nematic phase. A relatively sharp interface, formed between the two phases, can be controllably shifted by temperature-induced conversion between the phases. Once expelled, micelles do not remix into the nematic phase, whereas in the isotropic state their remixing takes place over several days. Temperature dependence of the linewidth of isotropic spectral component has been analyzed in terms of molecular reorientations mediated by translational displacements, assuming isotropically distributed directors of nanosized nematic domains. With our results, the existence of the proposed transparent nematic state cannot be completely ruled out. However, if present, the nematic order in such a phase must be extremely low.

Polymer mobilization and drug release during tablet swelling. A 1H NMR and NMR microimaging study

The objective of this study was to investigate the swelling characteristics of a hydroxypropyl methylcellulose (HPMC) matrix incorporating the hydrophilic drug antipyrine. We have used this matrix to introduce a novel analytical method, which allows us to obtain within one experimental setup information about the molecular processes of the polymer carrier and its impact on drug release. Nuclear magnetic resonance (NMR) imaging revealed in situ the swelling behavior of tablets when exposed to water. By using deuterated water, the spatial distribution and molecular dynamics of HPMC and their kinetics during swelling could be observed selectively. In parallel, NMR spectroscopy provided the concentration of the drug released into the aqueous phase. We find that both swelling and release are diffusion controlled. The ability of monitoring those two processes using the same experimental setup enables mapping their interconnection, which points on the importance and potential of this analytical technique for further application in other drug delivery forms.

1H and19F nuclear magnetic resonance microimaging of water and chemical distribution in soil columns

Nuclear magnetic resonance (NMR) microimaging is a noninvasive and nondestructive technique that has great potential for the study of soil processes. Hydrogen-1 NMR microimaging techniques were used to examine the distribution of water in four different soil cores. Fluorine-19 NMR microimaging is also used to study the transport of three model contaminants (hexafluorobenzene, sodium fluoride, and trifluralin) in soil columns. The 1H water distribution studies demonstrate that NMR microimaging can provide unique detail regarding the nature and location of water in soils. Image distortion (magnetic susceptibility) was observed for soil samples low in water (20-28% by weight) and that contained an iron content of 0.73 to 0.99%. Highly resolved images were obtained for the organic-rich soil (Croatan sample) and also facilitated the analysis of bound and unbound soil water through varying spin echo times. The contaminant studies with 19F NMR demonstrated that preferential flow processes can be observed in soil cores in as little as 16 h. Studies with hexafluorobenzene produced the highest quality images whereas the definition decreased over time with both trifluralin and sodium fluoride as the compounds penetrated the soil. Nonetheless, both 1H and 19F NMR microimaging techniques demonstrate great promise for studying soil processes.

A simple method to discriminate diapause from non-diapause pupae in large and small white butterflies, Pieris brassicae and P. rapae crucivora

Differences in colour and shape have been used to discriminate diapause pupae from non-diapause pupae in butterflies. In the present study, we describe a simple discrimination method based on differences in the specific gravity of diapause and non-diapause pupae of large and small white butterflies, Pieris brassicae and Pieris rapae crucivora. When put into water, diapause pupae sink to the bottom (specific gravity is above 1.0), whereas non-diapause pupae float on the surface (specific gravity is below 1.0). Nuclear magnetic resonance microimaging revealed that this difference in specific gravity is due to a difference in the volume of an internal cavity located between the thorax and the abdomen in the pupae. The cavity appears quite early in development. We also examined the cavity in pupae of the unrelated swallowtail butterfly, Papilio xuthus, and found a similar cavity. However, in this species, the cavity is very small and, as a consequence, non-diapause as well as diapause pupae sink in water.

Moisture movement and thickness swelling in oriented strandboard, part 1. Analysis using nuclear magnetic resonance microimaging

A procedure was developed to observe the water absorption characteristics of small specimens of oriented strandboard (OSB) and solid wood in a nuclear magnetic resonance (NMR) microimaging facility. The procedure allowed a specimen to be surrounded in water while remaining in a fixed position within the imaging coil, after which the water could be removed and the specimen immediately imaged. This technique permitted images of the specimen to be taken after a number of different periods of water soak without having to remove and replace the specimen, thus easily maintaining the same image location. Both the distribution of free water and its movement as a function of time were observed using this technique. Inter-strand voids within OSB were shown to be the main route for moisture movement through a specimen of this type. The influence of the inherent anisotropy of OSB on moisture movement was investigated by sealing selected surfaces of a specimen with silicone to observe moisture absorption in predetermined directions.

Simultaneous NMR microimaging of multiple single‐cell samples

AbstractNuclear magnetic resonance (NMR) microimaging experiments performed on small samples such as single cells typically require long data acquisition times and are usually carried out on high‐field systems with limited access time. When using small probe coils on millimetre (or smaller) size scale to study single cells, it is desirable to take advantage of the comparatively large volume of homogeneous static magnetic field within the magnet to increase the efficiency of NMR data acquisition. Here we describe the development of a four‐channel probehead that consists of four individual solenoid coils (length 2.8 mm, outer diameter 2 mm) stacked along the z axis. The probehead was designed for a 17.6 Tesla (750 MHz) wide‐bore magnet, and operates within a 40 mm inner diameter gradient set with maximum strength 1 Tesla per metre. We performed simulations to estimate the impact of shielding between the coils on the B1 field distribution. Results showed that for coil‐shield separations on the order of a coil radius, no significant change in B1 homogeneity is expected, and therefore coils were spaced accordingly. Coil isolation of greater than 35 dB was achieved for all coils, and the Q values were ∼160 unloaded and ∼60 loaded with 100 mM NaCl solution. Practical use of the probehead was demonstrated by obtaining simultaneously four three‐dimensional T1 maps of chemically fixed Xenopus laevis oocytes at a spatial resolution of 30 × 60 × 60 μm. The T1 values for the nucleus were ∼2.7 seconds, and the animal pole ∼1.2 seconds. © 2004 Wiley Periodicals, Inc. Concepts Magn Reson Part B (Magn Reson Engineering) 22B: 7–14, 2004.

Epicuticular wax of large and small white butterflies, Pieris brassicae and P. rapae crucivora: qualitative and quantitative comparison between diapause and non-diapause pupae.

We compared the quantity and quality of the epicuticular wax of diapause and non-diapause pupae in two closely related Pieris species, P. brassicae and P. rapae crucivora. Main components of their epicuticular wax were identified as hydrocarbons. In P. brassicae, more than 95% of hydrocarbons were saturated regardless of whether the pupae were in diapause or not. In P. rapae crucivora, 93% of hydrocarbons were saturated in non-diapause pupae whereas in diapause pupae 41% were saturated and 59% unsaturated. From measurements of body surface area by nuclear magnetic resonance microimaging, we calculated the average thickness of the wax layer. The thickness in diapause and non-diapause pupae of P. brassicae was 800 and 160 nm, respectively. In P. rapae crucivora, the thickness was 195 nm in diapause and 11 nm in non-diapause. This is the first report to clarify the compositional difference in epicuticular wax between diapause and non-diapause pupae.

Regional cerebral blood volume reduction in transgenic mutant APP (V717F, K670N/M671L) mice

Recent advance in nuclear magnetic resonance (NMR) microimaging has enabled in vivo cerebral blood volume (CBV) mapping with high spatial resolution. Using an intravascular susceptibility contrast agent and T 2-weighted magnetic resonance imaging (MRI) on a 9.4 T NMR microimager, the regional CBV was measured in mice as the transverse relaxation increase induced by the contrast agent. CBV maps in an Alzheimer’s disease mouse model at resting state were obtained and examined. Four-month-old male transgenic mutant APP (V717F, K670N/M671L) mice ( N = 10) and littermate wild-type controls ( N = 12) were used. Regional analysis of the multi-slice CBV maps revealed statistically significant CBV reductions among the APP mice in cerebral cortex (−9.29%, P = 0.0002), hippocampus (−4.22%, P = 0.02), and thalamus (−5.21%, P = 0.03), indicating an early change of microvasculature in these selected regions. No significant difference was found in olfactory bulb, pons, midbrain, superior colliculus, medulla, and cerebellum.

Nuclear magnetic resonance microimaging of mouse spinal cord in vivo

The spinal cord is the site of traumatic injuries, the devastating consequences of which constitute a public health problem in our societies. So far, there is no efficient repair therapeutic approach, and this is mainly due to the great difficulty for elaborating predictive experimental models of this pathology. Up to now, most pathophysiological studies were based on postmortem evaluation of the quantity and extent of the lesions, and their comparison in-between human and rodent specimen. Recent progresses of magnetic resonance imaging provide new tools to examine in vivo rodent central nervous system, and eventually to monitor the progression of lesions. However, up to now, mice spinal cord has been inaccessible to such studies, due to specific physiological characteristics and to the small size of the cord. In this study, the first diffusion-weighted images depicting the mouse thoracic spinal cord in vivo are shown. Motion-related artifacts are significantly reduced by respiratory gating using a dedicated sensor. By changing the direction of diffusion-sensitizing gradients, different contrasts were obtained that are compared with ex vivo MRI and histological preparations. In addition, preliminary results obtained on pathological cords are presented.