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SOURCE OF CELLS FOR BIOLOGICAL REPAIR OF THE INTERVERTEBRAL DISC



Abstract

Introduction: Degeneration of the intervertebral disc is characterised by loss of normal cell activity, disc matrix and loss of disc height. There is currently much interest in using cells to effect a biological repair in connective tissues, eg autologous chondrocyte implantation for cartilage repair. Intervertebral discs have a low cell density, with those cells present often being unhealthy and necrotic. Hence, identification of an alternative source of cells for autologous disc repair could be beneficial. Thus we have investigated other types of connective tissue cells to determine if they may be encouraged to undertake a disc cell phenotype.

Materials and Methods: Cells were enzymatically/mechanically extracted from bovine coccygeal discs (annulus and nucleus), skin, bone marrow, periosteum and tendon and the efficiency and proliferation rates assessed. Dermal fibroblasts and bone marrow cells were also grown in a 3D alginate system and compared to disc nucleus pulposus cells for phenotypic expression from 0–28 days. Cell phenotype was assessed via morphology, immunohistochemistry, Western blotting and RT-PCR for mRNA expression.

Results: All cell types could be extracted and proliferated in monolayer, with a flattened and fibroblast-like morphology. Proliferation was slowest for bone marrow cells (4 times slower than nucleus pulposus cells). Cells cultured in alginate became rounded with chondrocyte-like morphology. They remained viable for 4 weeks, but with little replication. Expression or production of proteoglycans, both aggrecan and the small proteoglycans (especially fibromodulin) and collagen types I, II and X was demonstrated for all cell types. There was, however, a difference in the timescale of production between some cell types.

Conclusions: Plasticity of different cell types is well known and the connective tissue cells investigated in this study are capable of responding to the environment in which they are cultured. They can synthesise matrix molecules typically produced by disc cells in vivo and hence warrant further investigation as a potential source of cells for biological repair of the intervertebral disc.

The abstracts were prepared by Editorial Secretary, Dr Charles Pither. Correspondence should be addressed to SBPR at the Royal College of Surgeons, 35–43 Lincoln’s Inn Fields, London WC2A 3PN