Tameshwar Ganesh1,2, Marvin Estrada3, James Duffin4, and Hai-Ling Margaret Cheng1,2,5,6
1Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada, 2Ted Rogers Centre for Heart Research, Translational Biology & Engineering Program, Toronto, ON, Canada, 3Lab Animal Services, Hospital for Sick Children, 4Anesthesia, University of Toronto, Canada, 5The Edward S. Rogers Sr. Department of Electrical & Computer Engineering, University of Toronto, Toronto, ON, Canada, 6Institute of Biomaterials & Biomedical Engineering, University of Toronto, Toronto, ON, Canada
Synopsis
The ability of blood vessels to modulate
vascular tone (i.e. blood volume) is an important normal physiological
function, which is compromised in multiple diseases, including diabetes and
heart diseases. To date, there is no
non-invasive imaging technique to assess dynamic microvascular blood volume
response. Herein we present a novel MRI technique to assess vasoactive response
in “near” real-time in extra-cranial tissues.
We employ a blood-pool agent Gadofosveset (Vasovist) together with a
variety of vasoactive gas stimuli. In-vivo results in kidney and liver
demonstrate we can visualize dynamic changes in gas-induced blood volume
changes, as confirmed on laser Doppler perfusion.
Target Audience
physiologists, radiologists, oncologistsPURPOSE
The integration of MR
imaging and a gas challenge stimulus (i.e. application of inspired gases
containing different levels of O2 and CO2) is a method
for non-invasively assessing cerebral vascular response in both health and
disease. The application of this approach to assess extracranial tissue is an
emerging research topic1-4, but these studies have shown that T2*
and T1 response to gas challenges has multiple contributions
in addition to that from blood volume. In this study, we demonstrate a novel
technique using long-life blood pool agent Gadofosveset (Vasovist) combined
with varying degrees of hypercapnia and hypoxia, for specific monitoring of vasoactive
perfusion changes in extracranial tissues.Methods
This study was approved by our institutional animal care
committee. A system consisting of a computer-controlled gas mixer, an
MR-compatible isoflurane vaporizer and ventilator was built to control the
delivery of precise levels of gases to small animals. Eight female adult
Sprague Dawley rats (Charles River Laboratories) weighing 250-300g were
anesthetized on 5% isoflurane, intubated with a 14-gauge angiocatheter, and
transferred to a water blanket maintained at 38°C. An endotracheal tube was connected to the gas delivery
system, and the animal was maintained on 2% isoflurane, resting supine in an
8-channel wrist coil and imaged on a 3-Tesla scanner (Achieva 3.0 T TX, Philips
Medical Systems). Vasovist (0.3mmol kg-1) was administered via the
tail vein, and 10 min sequential gas challenges beginning with normoxia (21% O2)
were applied. Four different types of gas challenge were studied: extreme
hypercapnia (20% CO2), mild hypercapnia (5% CO2), hypoxia
(12% O2), and hypercapnic hypoxia (20% CO2 +12%O2).
Four animals were used to study each level of gas stimulus. Coronal 3D T1-weighted
spoiled-gradient echo fat-suppressed images were acquired: TR=3.73 ms, TE=1.85
ms, FA=20°, NSA=3,
FOV=100mm, ten 3-mm slices, 0.7x0.7 mm
in-plane was acquired at 2 minute intervals for each gas challenge. MRI data
was analyzed using in-house software in Matlab (v.8.3). Regions of interest
were outlined on the liver, kidney cortex, and paraspinal muscle to obtain mean
signal intensity values. Changes in signal intensity from baseline were compared
across animals using one-way ANOVA and post-hoc Tukey-Kramer tests performed at
the 95% confidence level.
Real-time
relative perfusion measurements were obtained in the liver and kidney cortex on
seven female rats in response to the gas challenge regime using a laser Doppler
perfusion (LDP) (OxyFlo, Oxford Optronics Ltd., Oxford, UK) fiber-optic system.
The fiber optic probes were inserted below the renal capsule to measure renal cortical
perfusion and between the lobes of the liver to measure liver blood perfusion.Results
Fig. 1 shows relative changes in T1 signal in different abdominal
tissues across all animals for all gas challenge levels. As seen in Fig. 1, T1
signal decreased consistently for hypercapnic, hypoxic, and hypercapnic hypoxic
challenges when compared to baseline, with significance seen in the kidney
cortex. The hypercapnic reduction was sustained with addition of a hypercapnic
hypoxic stimulus, and these trends either from hypercapnia or hypercapnic
hypoxia were reversed upon the administration of mild hypercapnia (5% CO2).
Fig. 2 shows anatomical images of the kidney and changes in signal intensity in
all tissues in response to one of the gas sequences. This figure exemplifies
the large changes in signal obtained, particularly in the kidney cortex, for
visual real-time monitoring of vasoactive response. The underlying perfusion
changes observed on MRI were corroborated with LDP measurements (Fig. 3).Discussion
Using a long-life blood pool agent, we are able to visualize on MRI active
modulation of blood volume in the kidney and liver. The hypercapnic and
hypercapnic hypoxic mediated reduction in signal intensity and perfusion is
attributed to the vasoconstrictive effects of extreme levels of CO2
and O2 on the renal
vasculature, whilst mild hypercapnia resulted in vasodilatation. Comparison
between MR and LDP showed good agreement. For example, as seen in Figs. 2 and 3,
extreme hypercapnia resulted in a 35% reduction in the renal cortical signal
intensity, in good agreement with the 31% reduction in relative perfusion seen
on LDP. In contrast, mild hypercapnia led to vasodilatation, confirmed on LDP
and seen as signal enhancement on MRI.Conclusion
We have presented a novel imaging approach for the real-time assessment
of vasoactive response and, thereby, microvascular health of tissues. This
approach may be extended to other imaging modalities for the investigation of
microvessel-related diseases.Acknowledgements
T. Ganesh is supported by the Ontario Graduate Scholarship and NSERC PGS-D. H-L. M. Cheng is funded by the Heart & Stroke Foundation of Canada, NSERC Discovery Awards, the Canada Foundation for Innnovation, and the Ontario Research Funds.References
1.
Winter JD et al. Normal tissue
quantitative T1 and T2* MRI relaxation time responses to hypercapnic and
hyperoxic gases. Acad Radiol 201 1 ; 1 8(9);1 1 59-1 1 67.
2.
O’Connor JP et al. Organ-specific effects of
oxygen and carbogen gas inhalation on tissue longitudinal relaxation times.
Magn Reson Med 2007; 58(3):490-496.
3.
Pohlmann A et al. Detailing the relation
between renal T2* and renal tissue pO2 using an integrated approach of
parametric magnetic resonance imaging and invasive physiological measurements.
Invest Radiol 201 4;49(8):547-560.
4.
Ganesh T, Estrada M, Duffin J,
Cheng HL.
“T2* and T1 assessment of abdominal tissue response to graded hypoxia and hypercapnia
using a controlled gas mixing circuit for small animals,” Journal of Magnetic Resonance Imaging
44(2), 305-316, 2016.