Immunoselected STRO-3+mesenchymal precursor cells and restoration of the extracellular matrix of degenerate intervertebral discs

Laboratory investigation

Restricted access

Object

Chronic low-back pain of discal origin is linked strongly to disc degeneration. Current nonsurgical treatments are palliative and fail to restore the disc extracellular matrix. In this study the authors examined the capacity of ovine mesenchymal precursor cells (MPCs) to restore the extracellular matrix of degenerate discs in an ovine model.

Methods

Three adjacent lumbar discs of 24 adult male sheep were injected intradiscally with chondroitinase-ABC (cABC) to initiate disc degeneration. The remaining lumbar discs were used as normal controls. Three months after cABC injection, the L3–4 discs of all animals were injected with either a high dose (4 × 106 cells, in 12 sheep) or low dose (0.5 × 106 cells, in 12 sheep) of MPCs suspended in hyaluronic acid (HA). The adjacent L4–5 degenerate discs remained untreated; the L5–6 discs were injected with HA only. The animals were euthanized at 3 or 6 months after MPC injections (6 sheep from each group at each time point), and histological sections of the lumbar discs were prepared. Radiographs and MR images were obtained prior to cABC injection (baseline), 3 months after cABC injection (pretreatment), and just prior to necropsy (posttreatment).

Results

Injection of cABC decreased the disc height index (DHI) of target discs by 45%–50%, confirming degeneration. Some recovery in DHI was observed 6 months after treatment in all cABC-injected discs, but the DHI increased to within baseline control values only in the MPC-injected discs. This improvement was accompanied by a reduction in MRI degeneration scores. The histopathology scores observed at 3 months posttreatment for the high-dose MPC–injected discs and at 6 months posttreatment for the low-dose MPC–injected discs were significantly different from those of the noninjected and HA-injected discs (p <0.001) but not from the control disc scores.

Conclusions

On the basis of the findings of this study, the authors conclude that the injection of MPCs into degenerate intervertebral discs can contribute to the regeneration of a new extracellular matrix.

Abbreviations used in this paper:AF = anulus fibrosus; cABC = chondroitinase-ABC; DHI = disc height index; HA = hyaluronic acid (hyaluron); MPC = mesenchymal precursor cell; MSC = mesenchymal stem cell; NP = nucleus pulposus.

Article Information

Address correspondence to: Peter Ghosh, D.Sc., P.O. Box 35, Brookvale, New South Wales 2100, Australia. email: peter.ghosh@mesoblast.com.

Please include this information when citing this paper: published online March 9, 2012; DOI: 10.3171/2012.1.SPINE11852.

© AANS, except where prohibited by US copyright law.

Headings

Figures

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    Left: Baseline MR image obtained in a normal ovine lumbar spine showing the strong water signal for the NP of all discs prior to injection with cABC. Right: Pretreatment MR image obtained 3 months after cABC injection showing marked reduction in signal in the injected discs, confirming that cABC induced disc degeneration by initially degrading the water-binding proteoglycans of the NP. The nondegenerate control discs and the relative spinal levels of the 3 degenerate (cABC-injected) discs that were untreated or injected with either MPC+HA or HA alone are also indicated.

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    Graphs showing the mean MRI aggregate degeneration scores determined 3 months after cABC injection for the uninjected control discs and the cABC-injected discs that were to remain untreated (cABC), were to be injected with HA, or were to be injected with low-dose (A and C) or high-dose (B and D) MPC+HA. Before treatment, the values for all categories of cABC-injected discs were significantly different from those of the uninjected controls within each group. mths = months. Error bars indicate SEM. **p <0.001, ***p <0.0001 relative to control disc values.

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    Mean DHI values determined at baseline, pretreatment, and 3 and 6 months posttreatment for control discs (A), MPC+HA–injected discs (B), untreated cABC-injected discs (cABC) (C), and HA-injected discs (D) for spines from animals that received the low-dose MPC injections. Only the degenerate discs injected with MPCs (B) showed recovery in DHI after 6 months as indicated by the nonsignificant difference (NSD) in mean values relative to baseline. The DHI values for the untreated degenerate discs (C) and HA-injected degenerate discs (D) remained significantly different from baseline at 3 and 6 months posttreatment. Error bars indicate SEM. *p <0.01, ***p <0.0001 relative to baseline.

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    Mean DHI values determined at baseline, pretreatment, and 3 and 6 months posttreatment for control discs (A), MPC+HA–injected discs (B), untreated cABC-injected discs (“nil injected”) (C), and HA-injected discs (D) from animals that received the high-dose MPC injections. The degenerate discs injected with a high dose of MPCs (B) as well as those that were left untreated (C) showed a recovery in DHI to a value that was not significantly different from baseline. In contrast the DHI of the HA-injected discs (D) remained significantly different from baseline. Error bars indicate SEM. **p <0.001, ***p <0.0001 relative to baseline.

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    Mean MRI aggregate degeneration scores for control, untreated (cABC), HA-injected, and low- (A and C) and high-dose (B and D) MPC+HA–injected discs, 3 (A and B) and 6 (C and D) months posttreatment. The scores for the low-dose MPC–injected discs at 3 months and the scores for the high-dose MPC–injected discs at 3 and 6 months were all significantly lower than those of the untreated (cABC-injected) and HA-injected discs at those time points and were not significantly different from the control values. In contrast, the scores for the untreated and HA-injected discs were significantly higher than the control values. Error bars indicate SEM. *p <0.01, **p <0.001, ***p <0.0001 relative to control values; #p <0.01 relative to MPC-injected discs.

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    Criteria used to grade the histopathological changes observed in sections of discs from the various experimental groups. The system was based on a previous publication (Sasaki et al.42) that produces an overall score derived from pathological changes in all tissues of the disc. The photomicrographs show Alcian blue/neutral red–stained sections of the NP, AF, and cartilage endplate (CEP) of a normal disc highlighting the respective structural variations for these disc regions. BEP = bony endplate; PG = proteoglycan.

  • View in gallery

    Mean histopathology scores for control, untreated (cABC), HA-injected, and low- (A and C) and high-dose (B and D) MPC+HA–injected discs obtained 3 (A and B) and 6 (C and D) months posttreatment. The 3-month score for the high-dose (B) and 6-month score for the low-dose (C) MPC–injected discs were not significantly different from the scores of control discs but were lower than those of the untreated discs (p <0.001). The scores of the untreated (cABC) and HA-injected discs were significantly higher than the control disc scores at 3 and 6 months for both high- and low-dose MPC groups. Error bars indicate SEM. **p <0.001, ***p <0.0001 relative to control values; ##p <0.001 relative to the MPC-injected discs.

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