[Show abstract][Hide abstract] ABSTRACT: Due to their pure intracellular compartmentation, the translational diffusion of brain metabolites in vivo depends on the intracellular environment, including viscosity, molecular crowding and subcellular structures. However, as the diffusion time is increased, metabolites have enough time to significantly encounter cell boundaries, so that cell size and geometry are expected to strongly determine metabolites diffusion path. In the present work, diffusion-weighted nuclear magnetic resonance spectroscopy was used to investigate brain metabolites diffusion in vivo, at long and ultra-long diffusion times (from ~80ms to more than 1s), in a voxel with equal proportions of white and grey matter in macaque monkeys. No dramatic dependence of the ADC on the diffusion time was observed, suggesting that metabolites apparent diffusion is largely unrestricted over these time-scales. In an attempt to explain this stability and relate it to plausible cell geometries, data were analyzed with two simple geometrical models describing diffusion either in fibers such as axons, dendrites and astrocytic processes, or in closed cell bodies. Results support the idea that DW-MRS is sensitive to cell shape, and that a vast fraction of brain metabolites is diffusing in long fibers rather than being confined in cell bodies.
[Show abstract][Hide abstract] ABSTRACT: PEGylated polyester nanocapsules of perfluorooctyl bromide (PFOB) were surface-decorated with a RGD (Arginine-Glycine-Aspartic acid) peptide by either pre-functionalization or post-functionalization strategies using carbodiimide-assisted chemistry. Both strategies allowed successful linkage of RGD at the surface of nanocapsules with up to 600 to 950 peptide units per nanocapsule without modifying the encapsulation efficacy of PFOB used as the (19)F MRI imaging moiety. Cryo-Transmission Electron Microscopy images evidence that slight changes of the polymer used to form the capsule shell strongly influence nanocapsule morphology. While, the use of copolymer blends induces the formation of acorn morphologies, PLA-b-PEG-COOH leads to elongated and "tears of wine"-like nanoconstructs. In vivo evaluation in mice bearing CT26 tumors by (19)F MRI reveals no significant difference of accumulation between PEGylated and RGD-decorated nanocapsules obtained by the post-functionalization approach (highest RGD density/capsule).
European journal of pharmaceutics and biopharmaceutics: official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V 12/2013; · 3.15 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: (13)C nuclear magnetic resonance (NMR) spectroscopy is the method of choice for studying brain metabolism. Indeed, the most convincing data obtained to decipher metabolic exchanges between neurons and astrocytes have been obtained using this technique, thus illustrating its power. It may be difficult for non-specialists, however, to grasp thefull implication of data presented in articles written by spectroscopists. The aim of the review is, therefore, to provide a fundamental understanding of this topic to facilitate the non-specialists in their reading of this literature. In the first part of this review, we present the metabolic fate of (13)C-labeled substrates in the brain in a detailed way, including an overview of some general neurochemical principles. We also address and compare the various spectroscopic strategies that can be used to study brain metabolism. Then, we provide an overview of the (13)C NMR experiments performed to analyze both intracellular and intercellular metabolic fluxes. More particularly, the role of lactate as a potential energy substrate for neurons is discussed in the light of (13)C NMR data. Finally, new perspectives and applications offered by (13)C hyperpolarization are described.
[Show abstract][Hide abstract] ABSTRACT: Molecular imaging with magnetic resonance imaging (MRI) targeted contrast agents has emerged as a promising diagnostic approach in cancer research to detect associated biomarkers. In this work, the potential of (19)F MRI was investigated to detect angiogenesis with α(ν)β(3)-targeted perfluorooctylbromide nanoparticles (PFOB NP) in a U87 glioblastoma mouse model at 7 Tesla. Mice were injected intravenously with targeted or non-targeted NP and (19)F images were immediately acquired for 90 min using a PFOB-dedicated MRI sequence. Mice infused with targeted NP exhibited higher concentrations in tumors than mice of the control group, despite the presence of nonspecific signal originating from the blood. Imaging results were corroborated by histology and fluorescence imaging, suggesting specific binding of targeted NP to α(ν)β(3) integrin. Two other groups of mice were injected 24 h before imaging to allow blood clearance but no significant differences were found between both groups, probably due to a loss of specificity of PFOB NP. This is the first demonstration of the ability of (19)F MRI to detect α(ν)β(3)-integrin endothelial expression in brain tumors in vivo.
[Show abstract][Hide abstract] ABSTRACT: Translational displacement of molecules within cells is a key process in cellular biology. Molecular motion potentially depends on many factors, including active transport, cytosol viscosity and molecular crowding, tortuosity resulting from cytoskeleton and organelles, and restriction barriers. However, the relative contribution of these factors to molecular motion in the cytoplasm remains poorly understood. In this work, we designed an original diffusion-weighted magnetic resonance spectroscopy strategy to probe molecular motion at subcellular scales in vivo. This led to the first observation of anomalous diffusion, that is, dependence of the apparent diffusion coefficient (ADC) on the diffusion time, for endogenous intracellular metabolites in the brain. The observed increase of the ADC at short diffusion time yields evidence that metabolite motion is characteristic of hindered random diffusion rather than active transport, for time scales up to the dozen milliseconds. Armed with this knowledge, data modeling based on geometrically constrained diffusion was performed. Results suggest that metabolite diffusion occurs in a low-viscosity cytosol hindered by ∼2-μm structures, which is consistent with known intracellular organization.Journal of Cerebral Blood Flow & Metabolism advance online publication, 29 August 2012; doi:10.1038/jcbfm.2012.119.
Journal of cerebral blood flow and metabolism: official journal of the International Society of Cerebral Blood Flow and Metabolism 08/2012; · 5.46 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: PLGA-PEG nanocapsules containing a liquid core of perfluorooctyl bromide were synthesized by an emulsion-evaporation process and designed as contrast agents for (19)F MRI. Physico-chemical properties of plain and PEGylated nanocapsules were compared. The encapsulation efficiency of PFOB, estimated by (19)F NMR spectroscopy, is enhanced when using PLGA-PEG instead of PLGA. PLGA-PEG nanocapsule diameter, measured by Dynamic Light Scattering is around 120 nm, in agreement with Transmission Electron microscopy (TEM) observations. TEM and Scanning Electron Microscopy (SEM) reveal that spherical core-shell morphology is preserved. PEGylation is further confirmed by Zeta potential measurements and X-ray Photoelectron Spectroscopy. In vitro, stealthiness of the PEGylated nanocapsules is evidenced by weak complement activation. Accumulation kinetics in the liver and the spleen was performed by (19)F MRI in mice, during the first 90 min after intravenous injection. In the liver, plain nanocapsules accumulate faster than their PEGylated counterparts. We observe PEGylated nanocapsule accumulation in CT26 xenograft tumor 7 h after administration to mice, whereas plain nanocapsules remain undetectable, using (19)F MRI. Our results validate the use of diblock copolymers for PEGylation to increase the residence time of nanocapsules in the blood stream and to reach tumors by the Enhanced Permeation and Retention (EPR) effect.
[Show abstract][Hide abstract] ABSTRACT: Metabolic modeling of dynamic (13)C labeling curves during infusion of (13)C-labeled substrates allows quantitative measurements of metabolic rates in vivo. However metabolic modeling studies performed in the brain to date have only modeled time courses of total isotopic enrichment at individual carbon positions (positional enrichments), not taking advantage of the additional dynamic (13)C isotopomer information available from fine-structure multiplets in (13)C spectra. Here we introduce a new (13)C metabolic modeling approach using the concept of bonded cumulative isotopomers, or bonded cumomers. The direct relationship between bonded cumomers and (13)C multiplets enables fitting of the dynamic multiplet data. The potential of this new approach is demonstrated using Monte-Carlo simulations with a brain two-compartment neuronal-glial model. The precision of positional and cumomer approaches are compared for two different metabolic models (with and without glutamine dilution) and for different infusion protocols ([1,6-(13)C(2)]glucose, [1,2-(13)C(2)]acetate, and double infusion [1,6-(13)C(2)]glucose + [1,2-(13)C(2)]acetate). In all cases, the bonded cumomer approach gives better precision than the positional approach. In addition, of the three different infusion protocols considered here, the double infusion protocol combined with dynamic bonded cumomer modeling appears the most robust for precise determination of all fluxes in the model. The concepts and simulations introduced in the present study set the foundation for taking full advantage of the available dynamic (13)C multiplet data in metabolic modeling.
Neurochemical Research 04/2012; · 2.13 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Early diagnosis and follow-up of neurodegenerative diseases are often hampered by the lack of reliable biomarkers. Neuroimaging techniques like magnetic resonance spectroscopy (MRS) offer promising tools to detect biochemical alterations at early stages of degeneration. Intracellular pH, which can be measured noninvasively by (31)P-MRS, has shown variations in several brain diseases. Our purpose has been to evaluate the potential of MRS-measured pH as a relevant biomarker of early degeneration in Huntington's disease (HD). We used a translational approach starting with a preclinical validation of our hypothesis before adapting the method to HD patients. (31)P-MRS-derived cerebral pH was first measured in rodents during chronic intoxication with 3-nitropropionic acid (3NP). A significant pH increase was observed early into the intoxication protocol (pH=7.17±0.02 after 3 days) as compared with preintoxication (pH=7.08±0.03). Furthermore, pH changes correlated with the 3NP-induced inhibition of succinate dehydrogenase and preceded striatum lesions. Using a similar MRS approach implemented on a clinical MRI, we then showed that cerebral pH was significantly higher in HD patients (n=7) than in healthy controls (n=6) (7.05±0.03 versus 7.02±0.01, respectively, P=0.026). Altogether, both preclinical and human data strongly argue in favor of MRS-measured pH being a promising biomarker for diagnosis and follow-up of HD.
Journal of cerebral blood flow and metabolism: official journal of the International Society of Cerebral Blood Flow and Metabolism 02/2012; 32(5):771-9. · 5.46 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We have recently developed an optimized multi-spin echo (MSE) sequence dedicated to perfluorooctyl bromide (PFOB) imaging yielding an excellent sensitivity in vitro. The aim of the present study was to apply this sequence to quantitative measurements in the mouse liver and spleen after intravenous (i.v.) injection of PFOB emulsions. We first performed oxygenation maps 25.5 min after a single infusion of emulsion and, contrary to previous studies, shortly after injection. The signal-to-noise ratio (SNR) in the liver and spleen was as high as 45 and 120, respectively, for 3-min images with 11.7-μL pixels. Values of oxygen tension tended to be slightly higher in the spleen than in the liver. Dynamic biodistribution experiments were then performed immediately after intravenous (i.v.) injection of PFOB emulsions grafted with different quantities of polyethylene glycol (PEG) for stealth. Images were acquired every 7 min for 84 min and the SNR measured in the liver and spleen was at least four from the first time point. Uptake rates could be assessed for each PEG amount and, in spite of high standard deviations (SDs) owing to interanimal variability, our data confirmed that increasing quantities of PEG allow more gradual uptake of the emulsion particles by the liver and spleen. In conclusion, our method seems to be a powerful tool to non-invasively perform accurate in vivo quantitative measurements in the liver and spleen using (19)F MRI.
NMR in Biomedicine 09/2011; 25(4):654-60. · 3.45 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In the present work, the non-linear phase dispersion induced by slice selective frequency-swept pulses is analyzed, in order to assess NMR signal attenuation due to molecular diffusion during such pulses. In particular, theoretical considerations show that diffusion-weighting can be calculated based on the non-linear phase spatial derivative (i.e. the phase gradient), and that the phase of B(1) field at the instant of the flip does not contribute to phase scrambling and diffusion-weighting, yielding a simple analytical expressions. The theory is validated by confrontation with numerical simulations of the Bloch equations including diffusion, performed for a pair of hyperbolic secant pulses and a pair of CHIRP pulses. The simple though general conceptual framework developed here should be useful for the understanding and the exact calculation of diffusion-weighting in NMR sequences using frequency-swept pulses.
Journal of Magnetic Resonance 08/2010; 205(2):255-9. · 2.30 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In the present work, the NMR properties of perfluorooctylbromide are revisited to derive a high-sensitivity fluorine MRI strategy. It is shown that the harmful effects of J-coupling can be eliminated by carefully choosing the bandwidth of the 180 degrees pulses in a spin-echo sequence. The T(2) of the CF(3) resonance of the molecule is measured using a multispin-echo sequence and shown to dramatically depend on the interpulse delay. Following these observations, an optimized multispin-echo imaging sequence is derived and compared with short TE/pulse repetition time gradient echo and chemical shift imaging sequences. The unparalleled sensitivity yielded by the multispin-echo sequence is promising for future applications, in particular for targeted contrast agents such as perfluorooctylbromide nanoparticles.
Magnetic Resonance in Medicine 04/2010; 63(4):1119-24. · 3.27 Impact Factor