Age-related degeneration of lumbar intervertebral discs in rabbits revealed by deuterium oxide-assisted MRI V.Y.L. Leung, Ph.D., S.-C. Hung, M.Sc., L.-C.

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Age-related degeneration of lumbar intervertebral discs in rabbits revealed by deuterium oxide-assisted MRI V.Y.L. Leung, Ph.D., S.-C. Hung, M.Sc., L.-C. Li, B.Sc., E.X. Wu, Ph.D., K.D.K. Luk, F.R.C.S., D. Chan, Ph.D., K.M.C. Cheung, M.D. Osteoarthritis and Cartilage Volume 16, Issue 11, Pages 1312-1318 (November 2008) DOI: 10.1016/j.joca.2008.03.015 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 T2-weighted/STIR MRI on deuterium oxide dilutions. H2O:D2O mixture and fish oil capsule were scanned without (A) or with (B) STIR sequence. Signal of fish oil capsule under STIR was suppressed preferentially. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Calibration curves from H2O:D2O phantoms. The normalized mean density of the standards was measured and plotted against HDi (HDi assigned to the dilution ratios) to establish a calibration curve for each scan. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Disc signal extraction from MRI scan. (A) Each animal was scanned in sagittal plane in the presence of H2O:D2O dilution panel. A representative scan of 1-year-old rabbit lumbar spine is shown. (B) Serial images of the IVDs (black–white inverted) were extracted from the scan. The intensity and the area of signal were measured to determine HDi from calibration curve. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Comparison of calibrated and non-calibrated measurements. The means of total integrated density of L3/L4 level were plotted against the age of the animals (non-calibrated measurement). For calibrated measurement, total disc signals were determined from the HDi calibration curves and then normalized to the mean of 6 months non-calibrated data. For semi-calibrated measurement, the signals were determined by first normalizing the total integrated density of the discs using the 40% H2O phantom signal in the corresponding scans, and the resulting means were then normalized to the mean of 6 months non-calibrated data. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Change of total HDi in lumbar discs with age. Sagittal MRI scans of 0.5–3 years old rabbits were obtained. The total HDi values of T12/L1, L1/L2, L3/L4, and L5/L6 IVDs were computed and plotted against age. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Change of total disc signal using non-calibrated measurement. From the same scans as shown in Fig. 4, the raw values of total integrated density of the lumbar discs were plotted against age. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 Change of maximum HDi in lumbar discs with age. From the same scans as shown in Fig. 4, the maximum HDi values of the lumbar discs were computed and plotted against age. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 8 Variation of HDi in rabbits. The Var(HDi) of the discs was analyzed at specific disc level and age. Osteoarthritis and Cartilage 2008 16, 1312-1318DOI: (10.1016/j.joca.2008.03.015) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions