Expression and regulation of neurotrophins in the nondegenerate and degenerate human intervertebral disc.
BACKGROUND
The neurotrophins nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) have been identified in the human intervertebral disc (IVD) and have been implicated in the mechanisms associated with nerve ingrowth and nociception in degeneration of the IVD. The aim of the current study was to investigate an association between neurotrophin expression in the IVD and the severity of disc degeneration, including the effect of disc-related proinflammatory cytokines on neurotrophin and neuropeptide expression in cells derived from the human IVD.


METHODS
Immunohistochemical analysis was performed to examine the expression of NGF, BDNF and their high-affinity receptors Trk-A and Trk-B in human IVD samples, divided into three categories: non-degenerate, moderate degeneration and severe degeneration. In order to study the effect of disc-related cytokines on neurotrophin/neuropeptide gene expression, nucleus pulposus cells derived from non-degenerate and degenerate IVD samples were seeded in alginate and were stimulated with either IL-1beta or TNFalpha for 48 hours. RNA was extracted, cDNA was synthesised and quantitative real-time PCR was performed to examine the expression of NGF, BDNF and substance P.


RESULTS
Immunohistochemistry showed expression of NGF and BDNF in the native chondrocyte-like cells in all regions of the IVD and in all grades of degeneration. Interestingly only BDNF significantly increased with the severity of degeneration (P < 0.05). Similar expression was observed for Trk-A and Trk-B, although no association with disease severity was demonstrated. In cultured human nucleus pulposus cells, stimulation with IL-1beta led to significant increases in NGF and BDNF gene expression (P < 0.05). Treatment with TNFalpha was associated with an upregulation of substance P expression only.


CONCLUSIONS
Our findings show that both the annulus fibrosus and nucleus pulposus cells of the IVD express the neurotrophins NGF and BDNF, factors that may influence and enhance innervation and pain in the degenerate IVD. Expression of Trk-A and Trk-B by cells of the nondegenerate and degenerate IVD suggests an autocrine role for neurotrophins in regulation of disc cell biology. Furthermore, modulation of neurotrophin expression by IL-1beta and modulation of substance P expression by TNFalpha, coupled with their increased expression in the degenerate IVD, highlights novel roles for these cytokines in regulating nerve ingrowth in the degenerate IVD and associated back pain.
Purmessur
Devina
D
Faculty of Medical and Human Sciences, School of Clinical and Laboratory Sciences, The University of Manchester, Manchester, UK.
Freemont
Anthony J
AJ
Hoyland
Judith A
JA
D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism*; D016022:Case-Control Studies; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D007403:Intervertebral Disc / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D020932:Nerve Growth Factor / Q000378:metabolism*; D009414:Nerve Growth Factors / Q000378:metabolism*; D020917:Receptor, trkA / Q000378:metabolism; D020813:Receptor, trkB / Q000378:metabolism; D012720:Severity of Illness Index; D013122:Spinal Diseases / Q000378:metabolism* / Q000473:pathology; D013373:Substance P / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D015415:Biomarkers; D019208:Brain-Derived Neurotrophic Factor; D053583:Interleukin-1beta; D009414:Nerve Growth Factors; D014409:Tumor Necrosis Factor-alpha; D013373:Substance P; D020932:Nerve Growth Factor; D020917:Receptor, trkA; D020813:Receptor, trkB