Abstract

IntroductionThe aims of these studies were to identify the cytokine and chemokine expression profile of nucleus pulposus (NP) cells and to determine the relationships between NP cell cytokine and chemokine production and the characteristic tissue changes seen during intervertebral disc (IVD) degeneration.MethodsReal-time q-PCR cDNA Low Density Array (LDA) was used to investigate the expression of 91 cytokine and chemokine associated genes in NP cells from degenerate human IVDs. Further real-time q-PCR was used to investigate 30 selected cytokine and chemokine associated genes in NP cells from non-degenerate and degenerate IVDs and those from IVDs with immune cell infiltrates (‘infiltrated’). Immunohistochemistry (IHC) was performed for four selected cytokines and chemokines to confirm and localize protein expression in human NP tissue samples.ResultsLDA identified the expression of numerous cytokine and chemokine associated genes including 15 novel cytokines and chemokines. Further q-PCR gene expression studies identified differential expression patterns in NP cells derived from non-degenerate, degenerate and infiltrated IVDs. IHC confirmed NP cells as a source of IL-16, CCL2, CCL7 and CXCL8 and that protein expression of CCL2, CCL7 and CXCL8 increases concordant with histological degenerative tissue changes.ConclusionsOur data indicates that NP cells are a source of cytokines and chemokines within the IVD and that these expression patterns are altered in IVD pathology. These findings may be important for the correct assessment of the ‘degenerate niche’ prior to autologous or allogeneic cell transplantation for biological therapy of the degenerate IVD.

Highlights

  • The aims of these studies were to identify the cytokine and chemokine expression profile of nucleus pulposus (NP) cells and to determine the relationships between NP cell cytokine and chemokine production and the characteristic tissue changes seen during intervertebral disc (IVD) degeneration

  • A diverse cytokine and chemokine mRNA expression profile was identified in total cells isolated from human NP tissue specimens cDNA Low Density Array (LDA) was performed to assess the diversity and extent of cytokine- and chemokin-associated gene expression by cells isolated from NP tissue

  • An extensive gene expression profile was detected, including 15 cytokines and chemokines not previously identified within the IVD; IL-11, IL-16, IL-18, IL-23, Tumour necrosis factor (TNF)-β, OSM, leukaemia inhibitory factor (LIF), CCL8, CCL19, CCL20, CXCL2, CXCL3, CXCL5, CXCL6 and CX3CL1 (Figures 2 and 3)

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Summary

Introduction

The aims of these studies were to identify the cytokine and chemokine expression profile of nucleus pulposus (NP) cells and to determine the relationships between NP cell cytokine and chemokine production and the characteristic tissue changes seen during intervertebral disc (IVD) degeneration. IVDs comprise three distinct tissue regions; the endplates, the annulus fibrosus (AF) and the nucleus pulposus (NP). The NP is the site of specific characteristic tissue changes whereby the hydrated gelatinous ECM is condensed and replaced by a more fibrous structure [3]. These structural changes compromise the biomechanics of the IVD, and can lead to prolapse [4]

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