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Interleukin-1α treatment of meniscal explants stimulates the production and release of aggrecanase-generated, GAG-substituted aggrecan products and also the release of pre-formed, aggrecanase-generated G1 and m-calpain-generated G1-G2

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Abstract

Pro-inflammatory cytokines induce meniscal matrix degradation and inhibition of endogenous repair mechanisms, but the pathogenic mechanisms behind this are mostly unknown. Therefore, we investigated details of interleukin-1 (IL-1α)-induced aggrecan turnover in mature meniscal tissue explants. Fibro-cartilagenous disks (3 mm diameter × 1 mm thickness) were isolated from the central, weight-bearing region of menisci from 2-year-old cattle. After 3 or 6 days of IL-1α-treatment, GAG loss (DMMB assay), biosynthetic activity ([35SO4]-sulfate and [3H]-proline incorporation), gene expression (quantitative RT-PCR) and the abundance (zymography, Western blot) of matrix-degrading enzymes and specific aggrecan products were determined. Meniscal fibrocartilage had a 4-fold lower GAG content (per wet weight) than adjacent articular cartilage, and expressed MMPs-1, -2, -3 and ADAMTS4 constitutively, whereas ADAMTS5 m-RNA was essentially undetectable. Significant IL-1 effects were a decrease in biosynthetic activity, an increase in GAG release and in the expression/abundance of MMP-2, MMP-3 and ADAMTS4. Fresh tissue contained aggrecan core protein products similar to those previously described for bovine articular cartilage of this age. IL-1 induced the release of aggrecanase-generated CS-substituted products including both high (>250 kDa) and low molecular weight (about 75 kDa) species. TIMP-3 (but not TIMP-1 and -2 or a broad spectrum MMP inhibitor) inhibited IL-1-dependent GAG loss. In addition, IL-1 induced the release of preformed pools of three known G1-bearing products. We conclude that aggrecanases are responsible for IL-1-stimulated GAG release from meniscal explants, and that IL-1 also stimulates release of G1-bearing products, by a process possibly involving hyaluronan fragmentation.

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Acknowledgements

We thank Rita Kirsch, Elsbeth Schulz, Frank Lichte and Marianne Ahler for their technical support. We also thank the Hofschlachterei Muhs (Krummbek) and NFZ Norddeutsche Fleischzentrale GmbH for the utilization of the knee joints.

Note

The amino acid residue numbers used in this manuscript are the bovine aggrecan sequence starting with N-terminal 1MTTL (NCBI accession no. P13608).

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Authors and Affiliations

  1. Institute of Anatomy, Christian-Albrechts-Universität Kiel, Kiel, Germany

    Angelika K. Lemke, Henning Voigt, Rolf Mentlein, Jakob Fay, Michael Schünke & Bodo Kurz

  2. Rush University Medical Center, Chicago, IL, USA

    John D. Sandy

  3. Institute of Physiological Chemistry and Pathobiochemistry, University of Münster, Münster, Germany

    Rita Dreier

  4. Wyeth Research, Collegeville, PA, USA

    Jennifer H. Lee

  5. Massachussetts Institute of Technology, Cambridge, MA, USA

    Alan J. Grodzinsky

  6. Anatomisches Institut, CAU zu Kiel, Olshausenstr. 40, 24098, Kiel, Germany

    Bodo Kurz

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  1. Angelika K. Lemke
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  2. John D. Sandy
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  3. Henning Voigt
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  9. Michael Schünke
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  10. Bodo Kurz
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Corresponding author

Correspondence to Bodo Kurz.

Additional information

Angelika K. Lemke and John D. Sandy contributed equally to this paper.

The study was funded by the Endo-Stiftung, Stiftung des Gemeinnützigen Vereins der ENDO-Klinik e.V., Hamburg.

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Lemke, A.K., Sandy, J.D., Voigt, H. et al. Interleukin-1α treatment of meniscal explants stimulates the production and release of aggrecanase-generated, GAG-substituted aggrecan products and also the release of pre-formed, aggrecanase-generated G1 and m-calpain-generated G1-G2. Cell Tissue Res 340, 179–188 (2010). https://doi.org/10.1007/s00441-010-0941-4

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