Ahmed A Khalil1,2,3, Susanne Mueller1,4, Marco Foddis1, Janet Lips1, Ulrich Dirnagl1, Sebastian Temme5, Ulrich Floegel5, and Philipp Boehm-Sturm1,4
1Department of Experimental Neurology and Center for Stroke Research Berlin, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany, 2Berlin School of Mind and Brain, Humboldt-Universität zu Berlin, Berlin, Germany, 3Department of Neurology, Max Planck Institute for Human Cognitive and Brain Sciences, Leipzig, Germany, 4Cluster of Excellence NeuroCure and Charité Core Facility 7T Experimental MRIs, Charité - Universitätsmedizin Berlin, Berlin, Germany, 5Institute of Molecular Cardiology, Heinrich-Heine-University of Düsseldorf, Düsseldorf, Germany
Synopsis
We assessed tissue oxygenation with 19F-MRI using perfluorocarbon emulsions in a mouse model of vascular cognitive impairment. Brain T1 values were measured using a cryogenic 19F/1H coil at multiple timepoints before and after surgery to induce bilateral common carotid artery stenosis and converted to pO2. T1 values decreased with increasing concentration of inhaled oxygen, and we observed a decrease in tissue pO2 following surgery which gradually recovered over four weeks. 19F-MRI of perfluorocarbon emulsions can be used for long-term assessment of brain tissue pO2 in vivo and may be relevant for monitoring endogenous or induced regenerative processes.
Introduction
A central pathophysiological process in cerebrovascular
disorders is the disturbance of the delivery of oxygen and glucose to tissues.1 Assessing tissue oxygenation
is therefore relevant for understanding the natural history of acute and
chronic vascular brain diseases and potentially for monitoring the effects of
both novel and established treatments. Often, surrogate measures such as blood
flow are used for this purpose instead. These measures are useful, but their
relationship to tissue nutrient delivery is complex, making their interpretation
difficult and possibly explaining the limited impact they have had thus far on patient
management.2 The longitudinal relaxation
rate (R1) of perfluorocarbons increases linearly with increased molecular
oxygen concentration3, and this property has been
used to quantify tissue oxygenation, particularly in mouse models of tumors.4 In this study, we applied 19F-MRI using a cryogenic
coil (with an SNR 2-3 times higher than room-temperature mouse head surface
coils) to assess brain oxygenation changes in a mouse model of vascular
cognitive impairment. Methods
Seventeen C57/BL6 mice underwent stereotactic injection of Rhodamin-labeled perfluoro-15-crown-5-ether (PFCE) emulsion into the striatum and corpus callosum (1 µl/deposit). Subsets of the mice were each imaged 1 week before (baseline) as well as 24 hours, 1 week, and 4 weeks after global hypoperfusion was induced by wrapping 160 µm microcoils around both common carotid arteries (Figure 1).5 Imaging was performed under isoflurane anesthesia (1.5 – 2%, adjusted to maintain vital signs) on a 7T Bruker BioSpin scanner with a cryogenic 19F/1H surface coil. It consisted of T2-weighted TurboRARE 1H (TR = 3500 ms, TE = 33 ms, 6 averages, FOV [mm] = 19.2 x 19.2, matrix = 128 x 128, slice thickness = 1 mm, acquisition time = 5 min 36 s, RARE factor = 8) and 19F (TR = 1000 ms, TE = 5 ms, 128 averages, FOV [mm] = 19.2 x 19.2, matrix = 48 x 48, slice thickness = 2 mm, acquisition time = 6 min 24 s, RARE factor = 16) images as well as T1 mapping with a single-slice 19F flow-sensitive alternating inversion recovery echo planar imaging (FAIR-EPI) sequence with adiabatic excitation pulses (TR = 5000 ms, TE = 9 ms, inversion times [ms] = (25, 400,800, 1200, 1600), 20 averages, FOV [mm] =19.2 x 19.2, matrix = 48 x 48, slice thickness = 6 mm, acquisition time = 12 min 1 s). At the baseline timepoint, the T1 mapping was repeated with three different concentrations of inhaled oxygen (30%, 60%, and 100%). T1 fitting was performed using a three-parameter model that includes variations in inversion efficiency to account for B1+ inhomogeneity. Fitting was done both on a voxel-by-voxel basis and on the mean signal from a region-of-interest, defined by an SNR > 5 mask on the FAIR-EPI images. T1 values were converted to pO2 using an in vitro calibration curve.6Results
SNR measurements of the 19F FAIR-EPI images varied across
mice but were largely stable over time (Figure 2). As expected, T1 decreased
with increasing inhaled O2 concentrations (Figure 3). In the mice who received
all 4 scans, a drop in tissue pO2 was observed 24 hours after
inducing hypoperfusion, followed by a gradual recovery by week 4 (Figure 4). Discussion
Assessment of brain tissue oxygenation in vivo using water-soluble perfluorocarbon emulsions, which are more biocompatible than liquid perfluorocarbons, is feasible. Importantly, the stability of these agents allows measuring of changes over time in vivo, which is particularly relevant for studying longitudinal regenerative processes or monitoring the effects of treatments aimed at improving oxygen delivery. Although the use of a cryogenic coil improves SNR, other factors potentially affecting data quality, including injection technique and the concentration of perfluorocarbon injected, need to be taken into careful consideration. Our data suggest that in the bilateral carotid stenosis model of vascular cognitive impairment, tissue oxygenation resembles previously described changes in blood flow.7 These changes may be related to regenerative processes such as neovascularization and collateral blood flow recruitment that are seen in this model.7Conclusion
19F-MRI of perfluorocarbon emulsions using cryogenic 19F/1H
coils is a promising method for long-term monitoring of changes in tissue
oxygenation over time in vivo.Acknowledgements
This work
was supported by the Stiftung Charité (BIH_PRO_317), the Federal Ministry of
Education and Research (BMBF) (grant number 01EO0801, Center for Stroke
Research Berlin), and the Deutsche Forschungsgemeinschaft (DFG) (Excellence
Cluster NeuroCure, Excellence Cluster BrainLinks-BrainTools). References
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