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Published in: BMC Musculoskeletal Disorders 1/2008

Open Access 01-12-2008 | Research article

Bovine explant model of degeneration of the intervertebral disc

Authors: Sally Roberts, Janis Menage, Sarit Sivan, Jill PG Urban

Published in: BMC Musculoskeletal Disorders | Issue 1/2008

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Abstract

Background

Many new treatments for degeneration of the intervertebral disc are being developed which can be delivered through a needle. These require testing in model systems before being used in human patients. Unfortunately, because of differences in anatomy, there are no ideal animal models of disc degeneration. Bovine explant model systems have many advantages but it is not possible to inject any significant volume into an intact disc. Therefore we have attempted to mimic disc degeneration in an explant bovine model via enzymatic digestion.

Methods

Bovine coccygeal discs were incubated with different concentrations of the proteolytic enzymes, trypsin and papain, and maintained in culture for up to 3 weeks. A radio-opaque solution was injected to visualise cavities generated. Degenerative features were monitored histologically and biochemically (water and glycosaminoglycan content, via dimethylmethylene blue).

Results and Conclusion

The central region of both papain and trypsin treated discs was macro- and microscopically fragmented, with severe loss of metachromasia. The integrity of the surrounding tissue was mostly in tact with cells in the outer annulus appearing viable. Biochemical analysis demonstrated greatly reduced glycosaminoglycan content in these compared to untreated discs. We have shown that bovine coccygeal discs, treated with proteolytic enzymes can provide a useful in vitro model system for developing and testing potential new treatments of disc degeneration, such as injectable implants or biological therapies.
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Literature
1.
go back to reference Thomas J, Lowman A, Marcolongo M: Novel associated hydrogels for nucleus pulposus replacement. J Biomed Mater Res. 2003, 67: 1329-1337.CrossRef Thomas J, Lowman A, Marcolongo M: Novel associated hydrogels for nucleus pulposus replacement. J Biomed Mater Res. 2003, 67: 1329-1337.CrossRef
2.
go back to reference Meisel HJ, Siodla V, Ganey T, Minkus Y, Hutton WC, Alasevic O: Clinical experience in cell-based therapeutics: Disc chondrocyte transplantation – A treatment for degenerated or damaged intervertebral disc. Biomolecular Engineering. 2006, 24: 5-21.CrossRefPubMed Meisel HJ, Siodla V, Ganey T, Minkus Y, Hutton WC, Alasevic O: Clinical experience in cell-based therapeutics: Disc chondrocyte transplantation – A treatment for degenerated or damaged intervertebral disc. Biomolecular Engineering. 2006, 24: 5-21.CrossRefPubMed
3.
go back to reference Alini M, Eisenstein SM, Ito K, Little CB, Kettler A, Masuda K, Melrose J, Ralphs JR, Stokes IAF, Wilke H-J: Are animal models useful for studying human disc disorders/degeneration?. Eur Spine J. 2008, 17: 2-19.CrossRefPubMed Alini M, Eisenstein SM, Ito K, Little CB, Kettler A, Masuda K, Melrose J, Ralphs JR, Stokes IAF, Wilke H-J: Are animal models useful for studying human disc disorders/degeneration?. Eur Spine J. 2008, 17: 2-19.CrossRefPubMed
4.
go back to reference Oshima H, Ishihara H, Urban JPG, Tsuji H: The use of coccygeal discs to study intervertebral disc metabolism. J Orthop Res. 1993, 11: 332-338.CrossRefPubMed Oshima H, Ishihara H, Urban JPG, Tsuji H: The use of coccygeal discs to study intervertebral disc metabolism. J Orthop Res. 1993, 11: 332-338.CrossRefPubMed
5.
go back to reference Roberts S, Menage J: Microscopic Methods for the Analysis of Engineered Tissues. Methods in Molecular Biology: Biopolymer Methods in Tissue Engineering. Edited by: Hollander AP, Hatton PV. 2004, Totowa: Humana Press Inc, 238: 171-195.CrossRef Roberts S, Menage J: Microscopic Methods for the Analysis of Engineered Tissues. Methods in Molecular Biology: Biopolymer Methods in Tissue Engineering. Edited by: Hollander AP, Hatton PV. 2004, Totowa: Humana Press Inc, 238: 171-195.CrossRef
6.
go back to reference Farndale RW, Buttle DJ, Barrett AJ: Improved Quantitation and Discrimination of Sulphated Glycosaminoglycans by use of Dimethylmethylene Blue. Biochim Biophys Acta. 1986, 883: 173-177.CrossRefPubMed Farndale RW, Buttle DJ, Barrett AJ: Improved Quantitation and Discrimination of Sulphated Glycosaminoglycans by use of Dimethylmethylene Blue. Biochim Biophys Acta. 1986, 883: 173-177.CrossRefPubMed
7.
go back to reference Roberts S, Menage J, Duance VC, Wotton S, Ayad S: Collagen Types Around the Cells of the Intervertebral Disc and Cartilage End Plate: An Immunolocalization Study. Spine. 1991, 16: 1030-1038.CrossRefPubMed Roberts S, Menage J, Duance VC, Wotton S, Ayad S: Collagen Types Around the Cells of the Intervertebral Disc and Cartilage End Plate: An Immunolocalization Study. Spine. 1991, 16: 1030-1038.CrossRefPubMed
8.
go back to reference Ariga K, Yonenobu K, Nakase T, Hosono N, Okuda S, Meng W, Tamura Y, Yoshikawa H: Mechanical stress-induced apoptosis of endplate chondrocytes in organ-cultured mouse intervertebral discs. Spine. 2003, 28: 1528-1533.PubMed Ariga K, Yonenobu K, Nakase T, Hosono N, Okuda S, Meng W, Tamura Y, Yoshikawa H: Mechanical stress-induced apoptosis of endplate chondrocytes in organ-cultured mouse intervertebral discs. Spine. 2003, 28: 1528-1533.PubMed
9.
go back to reference Risbud MV, Di Martino A, Guttapalli A, Seghatoleslami R, Denaro V, Vaccaro A, Albert T, Shapiro I: Toward an Optimum System for Intervertebral Disc Organ Culture. Spine. 2006, 31: 884-890.CrossRefPubMed Risbud MV, Di Martino A, Guttapalli A, Seghatoleslami R, Denaro V, Vaccaro A, Albert T, Shapiro I: Toward an Optimum System for Intervertebral Disc Organ Culture. Spine. 2006, 31: 884-890.CrossRefPubMed
10.
go back to reference Risbud MV, Izzo MW, Adams CS, Arnold WW, Hillibrand AS, Vresilovic EJ, Vaccaro AR, Albert TJ, Shapiro IM: An organ culture system for the study of the nucleus pulposus: description of the system and evaluation of the cells. Spine. 2003, 28: 2652-2659.CrossRefPubMed Risbud MV, Izzo MW, Adams CS, Arnold WW, Hillibrand AS, Vresilovic EJ, Vaccaro AR, Albert TJ, Shapiro IM: An organ culture system for the study of the nucleus pulposus: description of the system and evaluation of the cells. Spine. 2003, 28: 2652-2659.CrossRefPubMed
11.
go back to reference Chiba K, Andersson GB, Masuda K, Momohara S, Williams JM, Thonar JM: A new Culture System to Study the Metabolism of the Intervertebral Disc in vitro. Spine. 1998, 23: 1821-1828.CrossRefPubMed Chiba K, Andersson GB, Masuda K, Momohara S, Williams JM, Thonar JM: A new Culture System to Study the Metabolism of the Intervertebral Disc in vitro. Spine. 1998, 23: 1821-1828.CrossRefPubMed
12.
go back to reference Lee CR, Iatridis JC, Poveda L, Alini M: In vitro organ culture of the bovine intervertebral disc-effects of vertebral endplate and potential for mechanobiology. Spine. 2006, 31: 515-522.CrossRefPubMed Lee CR, Iatridis JC, Poveda L, Alini M: In vitro organ culture of the bovine intervertebral disc-effects of vertebral endplate and potential for mechanobiology. Spine. 2006, 31: 515-522.CrossRefPubMed
13.
go back to reference Gantenbein B, Grunhagen T, Lee CR, van Donkelaar CC, Alini M, Ito K: An in vitro organ culturing system for intervertebral disc explants with vertebral endplates: a feasibilty study with ovine caudal discs. Spine. 2006, 31: 2665-2673.CrossRefPubMed Gantenbein B, Grunhagen T, Lee CR, van Donkelaar CC, Alini M, Ito K: An in vitro organ culturing system for intervertebral disc explants with vertebral endplates: a feasibilty study with ovine caudal discs. Spine. 2006, 31: 2665-2673.CrossRefPubMed
14.
15.
go back to reference Imai Y, Okuma M, An HS, Nakagawa K, Yamada M, Muehleman C, Thonar E, Masuda K: Restoration of disc height loss by recombinant human osteogenic protein-1 injection into intervertebral discs undergoing degeneration induced by an intradiscal injection of chondroitinase ABC. Spine. 2007, 32: 1197-1205.CrossRefPubMed Imai Y, Okuma M, An HS, Nakagawa K, Yamada M, Muehleman C, Thonar E, Masuda K: Restoration of disc height loss by recombinant human osteogenic protein-1 injection into intervertebral discs undergoing degeneration induced by an intradiscal injection of chondroitinase ABC. Spine. 2007, 32: 1197-1205.CrossRefPubMed
16.
go back to reference Norcross JP, Lester GE, Weinhold P, Dahners LE: An in vivo model of degenerative disc disease. J Orthop Res. 2003, 21: 183-185.CrossRefPubMed Norcross JP, Lester GE, Weinhold P, Dahners LE: An in vivo model of degenerative disc disease. J Orthop Res. 2003, 21: 183-185.CrossRefPubMed
17.
go back to reference Antoniou J, Mwale F, Demers CN, Beaudoin G, Goswami T, Aebi M, Alini M: Quantitative magnetic resonance imaging of enzymatically induced degradation of the nucleus pulposus of intervertebral discs. Spine. 2006, 31: 1547-1554.CrossRefPubMed Antoniou J, Mwale F, Demers CN, Beaudoin G, Goswami T, Aebi M, Alini M: Quantitative magnetic resonance imaging of enzymatically induced degradation of the nucleus pulposus of intervertebral discs. Spine. 2006, 31: 1547-1554.CrossRefPubMed
Metadata
Title
Bovine explant model of degeneration of the intervertebral disc
Authors
Sally Roberts
Janis Menage
Sarit Sivan
Jill PG Urban
Publication date
01-12-2008
Publisher
BioMed Central
Published in
BMC Musculoskeletal Disorders / Issue 1/2008
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/1471-2474-9-24

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