Stefan Zbyn1,2, Kai D. Ludwig1,2, Lauren Watkins3, Alexandra R. Armstrong4, Russell L. Lagore1,2, Amanda Nowacki1, Marc A. Tompkins5, Ferenc Toth4, Gregor Adriany1,2, Kevin G. Shea6, Garry Gold7, Armin M. Nagel8, Cathy S. Carlson4, Gregory J. Metzger1,2, and Jutta M. Ellermann1,2
1Center for Magnetic Resonance Research, University of Minnesota, Minneapolis, MN, United States, 2Department of Radiology, University of Minnesota, Minneapolis, MN, United States, 3Department of Bioengineering, Stanford University, Stanford, CA, United States, 4Department of Veterinary Clinical Sciences, University of Minnesota, St. Paul, MN, United States, 5Department of Orthopaedic Surgery, University of Minnesota, Minneapolis, MN, United States, 6Department of Orthopaedic Surgery, Stanford Children's Hospital, Palo Alto, CA, United States, 7Department of Radiology, Stanford University, Stanford, CA, United States, 8Institute of Radiology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany
Synopsis
Sodium imaging is quantitative technique sensitive to
changes in cartilage glycosaminoglycan content. Changes in cartilage matrix,
due to maturation or degeneration, may influence sodium relaxation times which can
lead to incorrect sodium concentration estimates when not addressed. This ex
vivo study employs pediatric knee specimens to evaluate the relationship
between sodium relaxation parameters and compositional changes in the developing
cartilage matrix. Our preliminary evaluation suggests that cartilage maturation
is accompanied by a decrease in sodium T1 and the short T2* component. Sodium
concentrations in studies comparing healthy, diseased or immature cartilage
should be corrected for possible changes in relaxation times.
INTRODUCTION:
The
health and functional properties of articular cartilage are related to its glycosaminoglycan
(GAG) concentration, which can be evaluated with sodium MRI.1,2 Most studies calculate
sodium concentrations assuming the same sodium relaxation times in healthy
and degenerated or osteoarthritic cartilage. However, previous results suggest
relaxation times may increase in degenerated cartilage 3 and thus lead to the incorrect
sodium concentration measurements when not addressed. Pediatric cartilage at
different developmental stages offers a natural, non-enzymatic model to study the
relationship between sodium relaxation parameters and compositional changes in
developing cartilage matrix. The aims of this ex vivo 10.5T study were to: (i) evaluate sodium relaxation times
in the articular-epiphyseal cartilage complex of pediatric knee specimens, (ii)
calculate hydration, collagen content and GAG content in cartilage biopsies
from these same specimens, (iii) evaluate associations between specimen age, biochemical
composition, and sodium relaxation parameters.METHODS:
Six intact knee joint specimens from pediatric donors (4 females,
2 males; all left knees; age, 3-51 months) were received frozen from Allosource
Inc. (Centennial, CO). All specimens were thawed, immersed in
perfluoropolyether, and measured at room temperature using a whole-body 10.5T
MRI system (Siemens Magnetom, Erlangen, Germany).4 Immediately after MRI,
osteochondral core biopsies (diameter, 6 mm) from trochlear ridge were
acquired, wrapped in saline-soaked gauze and frozen at -20°C.
For sodium imaging, a 3T birdcage knee coil (Siemens) was modified
and re-tuned to the 10.5T sodium frequency (118.25 MHz). B0 shimming
and proton imaging were performed using a pair of fractionated proton dipole
antennas 4 driven with equal RF power and phase, and positioned inside the
sodium knee coil. Water-selective 3D DESS images (TR/TE: 15/5ms, FA: 50°, isotropic
resolution: 0.4mm, TA: 14-22 min) were acquired for the segmentation of the articular-epiphyseal
cartilage complex. All sodium images were acquired using a density 3D radial
projection sequence (DA-RP).5 Sodium B1+ maps were calculated using a
dual-angle method. Five sets of inversion recovery prepared DA-RP sequences were
acquired for T1 mapping (TR: TI+130ms, TE: 0.3ms, TI: 1-180ms, FA: 90°, 4500
projections, readout: 10ms, nominal isotropic resolution: 3.0mm, TA: 10-23 min/sequence).
Four sets of multi-echo DA-RP sequences (32 TEs) were acquired for
bi-exponential T2* mapping (TR: 70ms, TEs: 0.3, 0.9, 1.7,…, 63 ms, FA: 70°, 9500
projections, readout: 4ms, nominal isotropic resolution: 4.0mm, TA: 11 min/sequence).
All sodium images were reconstructed in Matlab (Mathworks,
Natick, MA) using a non-uniform fast Fourier transform method and Hann filter.
T1 and T2* maps were calculated pixel-by-pixel using three-parametric
mono-exponential and five-parametric bi-exponential least square fitting of
sodium signal in Matlab, respectively (Fig.1).
T1, short (T2*SHORT) and long (T2*LONG) components of T2* relaxation, signal fraction
of short T2* component (T2*PART), and corresponding adjusted coefficient of
determination (R2) maps were calculated for all specimens (Fig.2). Semi-automated segmentation of femoral, tibial and patellar
cartilage in DESS images was performed in ITK-SNAP (Fig.2). Segmentation masks were down-sampled to the resolution of
sodium relaxation maps and the mean values for each articular-epiphyseal cartilage
compartment were obtained from each knee specimen in Matlab (Fig.3).
For biochemical evaluation, osteochondral
biopsies were thawed and, after removing bone tissue, cartilage cores were
divided into cross-sectional discs. Sample wet and dry weights were recorded
before further cartilage treatment. Proteoglycan and collagen content were
assessed from spectrophotometric evaluations of the 1,9-dimethylmethylene blue
and the O-hydroxyproline biochemical assays, respectively. The mean hydration,
collagen, and sulfated glycosaminoglycan content (sGAG) were calculated for
each core biopsy (Fig.4). Pearson’s
correlation coefficients (r) were calculated to evaluate association between
specimen age and biochemical and sodium relaxation parameters.RESULTS:
The
ranges of mean sodium relaxation times in all evaluated specimens were: T1= 22-31
ms, T2*SHORT= 0.6-1.8 ms, T2*LONG= 6.3-13.4 ms, T2*PART= 39-51% (Fig.3). High adjusted R2 values suggest
excellent performance of fitting models (Fig.3).
Specimen age was strongly correlated with cartilage hydration (r= -0.943) and
collagen content (r= 0.741), but not with sGAG content (Figs.4,5). Specimen age also showed strong correlation to T2*SHORT (r=
-0.731), medium correlation to T1 (r= -0.670) and T2*PART (r= 0.504), and no
correlation to T2*LONG (Fig.5).DISCUSSION:
The
preliminary results of the present pediatric articular cartilage study suggest
a decrease in sodium T1 and T2*SHORT relaxation times and an increase in T2*PART
with increasing age and skeletal maturation. Presented sodium T1 times are longer than previously reported T1 values in
adult cartilage ranging from 18 to 21 ms.6 Observed changes are consistent with a physiologic increase in
collagen content and organization, and a decrease in water content and GAG
content during cartilage maturation.7,8 Our initial biochemical results from
biopsies also suggest a trend towards a decrease in cartilage hydration and an increase
in collagen content with increasing specimen age.CONCLUSION:
Sodium
relaxation times may change as a result of compositional and structural changes
in the cartilage matrix caused by tissue maturation or degeneration. Assuming T2*
parameters form the youngest and oldest specimen in the study, the difference
between sodium concentrations calculated with and without correction for T2* was
more than 11% for TE of 0.3 ms and more than 17% for more frequently used TE of
0.5 ms. Studies comparing results from healthy and degenerated or immature cartilage
should account for changes in sodium relaxation parameters when evaluating sodium
concentrations.Acknowledgements
This study was supported by the NIH grants R01 AR070020,
P41 EB027061, and S10 RR029672.References
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