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An in vitro tissue model to study the effect of age on nucleus pulposus cells

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Abstract

Differentiation between age (physiological) and disease-induced changes in the nucleus pulposus will facilitate our understanding of the mechanism(s) leading to the development of degenerative disc disease. The aim of this study was to develop an in vitro model that would allow the study of age-induced alterations of cell function in nucleus pulposus. Nucleus pulposus (NP) cells were isolated from intervertebral discs obtained from either calves (<9 months) or cows (>18 months). The cells were placed in culture and grown for 19 days. Although nucleus pulposus tissue was formed by the cells of the two different ages the more mature (older) cells formed less tissue as determined histologically by light microscopy. This was confirmed biochemically as the wet weight and proteoglycan content of the tissue formed by the older cells were significantly less than that of the younger tissue. The older cells accumulated less proteoglycans as determined by quantifying radioisotope incorporation. The older cells showed lower constitutive gene expression of collagen type II and aggrecan whereas collagen type I and link protein levels were similar to those of the younger cells. Metalloprotease (MMP) 13 gene and protein expression increased with age. There was no change in the levels of gene expression of MMP 2 and TIMP 1, 2, or 3 with age. Cells obtained from NP tissue harvested from younger or mature animals showed both genotypic and phenotypic differences in vitro that resulted in the inability of the older cells to reconstitute their extracellular matrix to the same extent as the younger cells. This suggests that this in vitro NP tissue model will be suitable to determine the mechanism(s) regulating age-induced changes.

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Acknowledgments

CS and DH were supported by fellowships from NSERC. We would like to thank Dr Jason Hong for making the CPP, Jennifer Yip for technical assistance and Marie Maguire for secretarial assistance. We would also like to thank Mr Harry Bojarski and Ryding-Regency Meat Packers, Toronto for the tissues. This work was supported by a grant from The Arthritis Society and CIHR-NSERC Collaborative Health Program.

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Authors and Affiliations

  1. CIHR-BioEngineering of Skeletal Tissues Team, Department of Laboratory Medicine and Pathobiology, Mount Sinai Hospital, University of Toronto, 600 University Avenue, Suite 6-500, M5X 1G5, Toronto, ON, Canada

    R. A. Kandel, D. Hamilton, C. Séguin, S.-Q. Li & C. Arana

  2. Faculty of Dentistry, University of Toronto, Toronto, ON, Canada

    R. Pilliar

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  1. R. A. Kandel
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  2. D. Hamilton
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  3. C. Séguin
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  4. S.-Q. Li
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  5. C. Arana
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  6. R. Pilliar
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Correspondence to R. A. Kandel.

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Kandel, R.A., Hamilton, D., Séguin, C. et al. An in vitro tissue model to study the effect of age on nucleus pulposus cells. Eur Spine J 16, 2166–2173 (2007). https://doi.org/10.1007/s00586-007-0467-y

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  • DOI: https://doi.org/10.1007/s00586-007-0467-y

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