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Development of an intact intervertebral disc organ culture system in which degeneration can be induced as a prelude to studying repair potential

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Abstract

The present work describes a novel bovine disc organ culture system with long-term maintenance of cell viability, in which degenerative changes can be induced as a prelude to studying repair. Discs were isolated with three different techniques: without endplates (NEP), with bony endplates (BEP) and with intact cartilage endplates (CEP). Swelling, deformation, and cell viability were evaluated in unloaded cultures. Degeneration was induced by a single trypsin injection into the center of the disc and the effect on cell viability and matrix degradation was followed. Trypsin-treated discs were exposed to TGFβ to evaluate the potential to study repair in this system. NEP isolated discs showed >75% maintained cell viability for up to 10 days but were severely deformed, BEP discs on the other hand maintained morphology but failed to retain cell viability having only 27% viable cells after 10 days. In CEP discs, both cell viability and morphology were maintained for at least 4 weeks where >75% of the cells were still viable. To mimic proteoglycan loss during disc degeneration, a single trypsin injection was administered to the center of the disc. This resulted in 60% loss of aggrecan, after 7 days, without affecting cell viability. When TGFβ was injected to validate that the system can be used to study a repair response following injection of a bio-active substance, proteoglycan synthesis nearly doubled compared to baseline synthesis. Trypsin-treated bovine CEP discs therefore provide a model system for studying repair of the degenerate disc, as morphology, cell viability and responsiveness to bio-active substances were maintained.

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Acknowledgments

This work was supported by financial support from AOSpine grant number AOSBR-07-07, the Canadian Arthritis Network and the Shriners of North America.

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Authors and Affiliations

  1. Orthopaedic Research Laboratory, McGill University, 687 Pine Avenue West, Room L4.70, Montreal, QC, H3A 1A1, Canada

    Bernice Jim, Thomas Steffen, Janet Moir & Lisbet Haglund

  2. Shriners Hospital for Children, McGill University, 1529 Cedar Avenue, Montreal, QC, H3G 1AG, Canada

    Peter Roughley

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  1. Bernice Jim
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  2. Thomas Steffen
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  3. Janet Moir
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  4. Peter Roughley
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  5. Lisbet Haglund
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Correspondence to Lisbet Haglund.

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Jim, B., Steffen, T., Moir, J. et al. Development of an intact intervertebral disc organ culture system in which degeneration can be induced as a prelude to studying repair potential. Eur Spine J 20, 1244–1254 (2011). https://doi.org/10.1007/s00586-011-1721-x

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  • DOI: https://doi.org/10.1007/s00586-011-1721-x

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