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Angela Ariza de Schellenberger,1 Harald Kratz,1 Tracy D Farr,2,3 Norbert Löwa,4 Ralf Hauptmann,1 Susanne Wagner,1 Matthias Taupitz,1 Jörg Schnorr,1 Eyk A Schellenberger1 1Department of Radiology, 2Department of Experimental Neurology, Center for Stroke Research Berlin, Charité – Universitätsmedizin Berlin, Berlin, Germany; 3School of Life Sciences, University of Nottingham, Medical School, Nottingham, UK; 4Department of Biomagnetic Signals, Physikalisch-Technische Bundesanstalt Berlin, Berlin, Germany Abstract: Sensitive cell detection by magnetic resonance imaging (MRI) is an important tool for the development of cell therapies. However, clinically approved contrast agents that allow single-cell detection are currently not available. Therefore, we compared very small iron oxide nanoparticles (VSOP) and new multicore carboxymethyl dextran-coated iron oxide nanoparticles (multicore particles, MCP) designed by our department for magnetic particle imaging (MPI) with discontinued Resovist® regarding their suitability for detection of single mesenchymal stem cells (MSC) by MRI. We achieved an average intracellular nanoparticle (NP) load of >10 pg Fe per cell without the use of transfection agents. NP loading did not lead to significantly different results in proliferation, colony formation, and multilineage in vitro differentiation assays in comparison to controls. MRI allowed single-cell detection using VSOP, MCP, and Resovist® in conjunction with high-resolution T2*-weighted imaging at 7 T with postprocessing of phase images in agarose cell phantoms and in vivo after delivery of 2,000 NP-labeled MSC into mouse brains via the left carotid artery. With optimized labeling conditions, a detection rate of ~45% was achieved; however, the experiments were limited by nonhomogeneous NP loading of the MSC population. Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP-uptake-dependent biocompatibility studies and cell detection by MRI and future MPI. Additionally, using a 7 T MR imager equipped with a cryocoil resulted in approximately two times higher detection. In conclusion, we established labeling conditions for new high-relaxivity MCP, VSOP, and Resovist® for improved MRI of MSC with single-cell sensitivity. Keywords: magnetic field microdistortions, single-cell imaging, mesenchymal stem cells, VSOP, MCP, Resovist®

Susanne Wagner,1 Jörg Schnorr,1 Antje Ludwig,2 Verena Stangl,2 Monika Ebert,1 Bernd Hamm,1 Matthias Taupitz11Department of Radiology, Section of Experimental Radiology, Charité – Universitätsmedizin Berlin, Campus Charité Mitte, and Campus Benjamin Franklin, Berlin, Germany; 2Department of Cardiology, Section of Experimental Cardiology, Charité – Universitätsmedizin Berlin, Campus Charité Mitte, Berlin, GermanyObjective: To evaluate the suitability of citrate-coated very small superparamagnetic iron oxide particles (VSOP) as a contrast agent for identifying inflammation in atherosclerotic lesions using magnetic resonance imaging (MRI).Methods and results: VSOP, which have already been evaluated as a blood pool contrast agent for MR angiography in human clinical trials, were investigated in Watanabe heritable hyperlipidemic rabbits to determine to what extent their accumulation in atherosclerotic lesions is a function of macrophage density and other characteristics of progressive atherosclerotic plaques. In advanced atherosclerotic lesions, a significant MRI signal loss was found within 1 hour after intravenous administration of VSOP at the intended clinical dose of 0.05 mmol Fe/kg. Histological examinations confirmed correlations between the loss of MRI signal in the vessel wall and the presence of Prussian blue-stained iron colocalized with macrophages in the plaque cap, but surprisingly also with calcifying microvesicles at the intimomedial interface. Critical electrolyte magnesium chloride concentration in combination with Alcian blue stain indicates that highly sulfated glycosaminoglycans are a major constituent of these calcifying microvesicles, which may serve as the key molecules for binding VSOP due to their highly complexing properties.Conclusion: Calcifying microvesicles and macrophages are the targets for intravenously injected VSOP in atherosclerotic plaques, suggesting that VSOP-enhanced MRI may render clinically relevant information on the composition and inflammatory activity of progressive atherosclerotic lesions at risk of destabilization.Keywords: atherosclerosis, inflammation, magnetic resonance imaging, iron oxide nanoparticles, glycosaminoglycans, calcifying microvesicles

We investigated the 2H nuclear magnetic resonance (NMR) line shape of deuterated benzene and hexamethylbenzene as guest molecules in organic glasses in the temperature range of 10–150 K. A broad distribution G(ln τ) of correlation times determines the slowing down of the molecular reorientation around the sixfold symmetry axis of the guests. The line shape is described by a superposition of temperature-dependent fractions F(T) of only two subspectra corresponding to fast and slowly rotating molecules; no spectra characteristic for intermediate mobility as found in crystal matrices are observed. Assuming a thermally activated motional process, the temperature dependence of G(ln τ) comes from a temperature-independent distribution of activation energies g(E). In this case, the derivative of the fraction dF(T)/dT yields directly the distribution g(E). Using this method an asymmetric distribution g(E) with its maximum at the low energy side is found for the glasses. While the general shape of g(E) is similar for different matrices, the mean activation energy differs significantly. We used the same approach to discuss similar ‘‘two-phase’’ spectra for the isotropic reorientation of toluene in polystyrene below the glass transition of the mixed system. Here, an alternative explanation is offered considering a distribution of glass transition points Tg for the dynamics of toluene in the mixed system. [ABSTRACT FROM AUTHOR]

Deuterated hexamethylbenzene (HMB) is used as a probe molecule for 2H NMR studies of the crystalline state of hexachlorobenzene and of several organic glasses. By measuring the spin–lattice relaxation and the line shape in the temperature range of 4–300 K the dynamical parameters of the molecular reorientation are investigated. For the system HMB/hexachlorobenzene, we find exponential relaxation and for the corresponding T1 an increase of its activation energy by a factor of 2 in comparison to the neat HMB. A homogeneous mixing of the guest and host molecules is found at least for guest concentrations up to 7%. In contrast, nonexponential spin–lattice relaxation is characteristic for all glass matrices, indicating motional heterogeneities. A log–Gauss distribution for the corresponding motional correlation times gives a good fit of the data. Its width parameter decreases linearly with temperature, while the mean correlation times are described by an Arrhenius law. The mean activation energy is reduced by a factor of about 3.5 as compared to neat HMB, demonstrating a loose packing of the molecules in the glass matrices. [ABSTRACT FROM AUTHOR]