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Skeletal keratan sulphate chains isolated from bovine intervertebral disc may terminate in alpha(2----6)-linked N-acetylneuraminic acid.

Peptido-keratan sulphate fragments were isolated from the nucleus pulposus of bovine intervertebral discs (2-year-old animals) after digestion with chondroitin ABC lyase followed by digestion with diphenylcarbamoyl chloride-treated trypsin of A1D1 proteoglycans and gel-permeation chromatography on Sepharose CL-6B. The peptido-keratan sulphate fragments were subjected to alkaline borohydride reduction. The reduced chains were treated with keratanase in the presence of the sialidase inhibitor 2,3-dehydro-2-deoxy-N-acetylneuraminic acid, and the digest was subjected to alkaline borohydride reduction. This produced oligosaccharides with galactitol at their reducing ends. This reduced digest was chromatographed on a Nucleosil 5 SB anion-exchange column and individual oligosaccharides were isolated. One of these was shown by 600 MHz 1H-n.m.r. spectroscopy to have the following structure: NeuAc alpha 2-6Gal beta 1-4GlcNAc(6-SO4)beta 1-3Gal-ol The structure of this oligosaccharide shows that keratan sulphate chains from bovine intervertebral disc have non-reducing termini with N-acetylneuraminic acid linked alpha(2----6) as well as alpha(2----3) to an unsulphated galactose.

Animals↗

MRI classification of lumbar herniated intervertebral disc.

A consecutive series of 28 patients with lumbar herniated intervertebral disc (HIVD) who exhibited positive magnetic resonance imaging (MRI) findings was analyzed and compared with operative findings to define the type of disc herniation. This prospective study showed that the overall accuracy of MRI in predicting the types of HIVD was 80.6%. Proton density image was the most helpful in differentiating protruded discs from extruded ones. Disruption of "posterior longitudinal ligament line," defined on T2-weighted sagittal image, was reliable in differentiating a transligamentous disc from subligamentous one. Sequestered disc could be more accurately diagnosed with Gadolinium-DTPA enhancement, which showed anterior rim enhancement of the sequestered portion on T1-weighted axial image.

Adolescent↗

Degenerative changes in the intervertebral discs of the lumbar spine and their sequelae.

Careful pathological examination of lumbar spines removed at autopsy has shown that degenerative changes are present in the intervertebral discs of all subjects by middle age. The degenerative changes are more marked and occur at an earlier age when evidence of vertical or posterior disc prolapse is also present. Degenerative changes in the discs are always accompanied by osteophyte formation on the margins of the vertebral bodies and remodelling changes in the apophyseal joints. There is a direct relationship between the degree of disc degeneration, marginal osteophyte formation on vertebral bodies, and apophyseal joint changes, which suggests that disc degeneration is the primary event leading to the clinical condition of 'degenerative spondylosis'.

Adolescent↗

2002 SSE Award Competition in Basic Science: expression of major matrix metalloproteinases is associated with intervertebral disc degradation and resorption.

During the process of degeneration, the intervertebral disc (IVD) shows a progressive and significant reduction in height due to tissue resorption. Intradiscal clefts and tears are major hallmarks of disc degeneration. Matrix-degrading enzymes such as matrix metalloproteinases (MMPs) are assumed to play a pivotal role in disc tissue degradation and resorption. The objective of this study was therefore to investigate the potential role of MMPs in extracellular matrix degradation leading to disc degeneration. This study was conducted on 30 formalin-fixed and EDTA-decalcified complete cross-sections of lumbar IVDs from cadavers of individuals aged between 0 and 86 years. Tissue sections were used for the immunolocalization of MMPs-1, -2, -3 and -9. The number of labeled cells was assessed by morphometric analyses, and was statistically correlated with the formation of clefts and tears, cellular proliferation, granular matrix changes and mucous degeneration. Furthermore, 30 disc specimens obtained during spinal surgery were used for in situ hybridization of MMP-2 and -3-mRNA. In addition, the enzymatic gelatinolytic activity was determined by in situ zymography in autopsy material. Immunohistochemistry showed the intradiscal expression of all four MMPs, which was confirmed by in situ hybridization, providing clear evidence for the synthesis of the enzymes within nucleus pulposus and annulus fibrosus cells. Gelatinolytic enzymatic activity was verified by in situ zymography. IVDs from infants and young adolescents remained almost completely unlabeled for all MMPs tested, while more MMPs-1 and -3 were seen in disc cells of younger adults than in those of a more advanced age; MMP-2 remained unchanged over the adult age periods, and MMP-9 was expressed in only relatively few cells. This pattern significantly correlated with the occurrence of clefts and tears. This correlation was strongest for MMP-1 ( P<0.0001), MMP-2 ( P<0.0017) and MMP-3 ( P<0.0005) in the nucleus, and MMP-1 ( P<0.0001) and MMP-2 ( P<0.038) in the annulus. In parallel, the proliferation of disc cells and matrix degeneration (granular changes and mucous degeneration) were related to MMP expression. Likewise, enzymatic activity was seen in association with cleft formation. Our data suggest that major MMPs play an important role in the degradation of the IVD. This is evidenced by the high correlation of MMP expression with the formation of clefts and tears. These findings implicate a leading function for MMPs in IVD degeneration resulting in the loss of normal disc function, eventually leading to low-back pain.

Adolescent↗

Cells from different regions of the intervertebral disc: effect of culture system on matrix expression and cell phenotype.

STUDY DESIGN: This study examined how the culture system and region of cellular origin affect disc cell morphology and extracellular matrix production. OBJECTIVE: To determine the role of the cell populations in the different regions of the adult intervertebral disc in maintaining gradients in composition across the disc. SUMMARY OF BACKGROUND DATA: It is not known whether the steep profiles in composition across the intervertebral disc are maintained by distinct cell populations or whether differences in cell metabolism are determined by changes in the physical environment across the disc. Very little information exists on the matrix produced by cells from the mature, non-notochordal nucleus pulposus. METHODS: Cells were extracted from articular cartilage, nucleus pulposus, and the inner and outer anulus fibrosus of caudal discs from 18- to 24-month-old steers cultured in alginate or collagen gels or in monolayer. The effect of culture system and cell origin on cell morphology and matrix synthesis was measured using 35S-sulphate labeling and indirect immunolocalization. RESULTS: Distinct morphologic differences between cells from different regions cultured in monolayer were retained through two passages. The rate of sulfate incorporation varied with cell type. Immediately after isolation, it was two- to threefold greater for nucleus cells than for cells from the disc inner anulus or articular cartilage. The rate was lowest for outer anulus cells. It also varied with culture system. For all cell types, the incorporation rate was highest in alginate and lowest in monolayer. Immunolocalization showed that nucleus cells stained strongly for all proteoglycan epitopes, whereas outer anulus cells stained least and in monolayer produced little proteoglycan. CONCLUSIONS: The disc has at least three distinct cell populations, which differ in morphology and in amount and type of matrix they produce. Cells from mature nucleus pulposus produced sulfated glycosaminoglycans at a high rate in contrast to reported results for notochordal nucleus cells. Alginate, although an appropriate culture system for inner anulus and nucleus cells, may not be a suitable medium for outer anulus cells.

Animals↗

Quantitative analysis of diurnal variation in volume and water content of lumbar intervertebral discs.

The Cavalieri method of modern design stereology has been used in combination with magnetic resonance imaging (MRI) to obtain unbiased estimates of the volume of the lumbar intervertebral discs immediately at the end of a day of normal activity and again following a night's rest. In addition, pixel-by-pixel mapping of the T2-relaxation time has been used to characterize objectively the tissues of the intervertebral discs. The mean increase in height of seven female subjects of average age 21 years (range 19-23 years) measured with a stediometer was 19.3 mm (range 8-26 mm). Image analysis showed that the mean overnight increase in volume of lumbar discs was 1300 mm3 (range 100-2700 mm3). The increase in volume of the disc was accompanied by an increase in the T2-relaxation time of the nucleus pulposus. This suggests that the change in disc volume is most probably caused by a preferential increase in the water content of the nucleus pulposus.

Adult↗

The qualitative assessment of glycosaminoglycans in the canine intervertebral disc using a critical electrolyte concentration staining technique.

Through the use of a critical electrolyte concentration staining technique, the glycosaminoglycans (GAGs) in the nucleus pulposus region of the canine intervertebral disc were arbitrarily identified as "hyaluronic acid", chondroitin sulphate and keratan sulphate. Approximate estimates of GAG concentration could be qualitatively deduced by determining the appropriate GAG "ALCIANOPHILIC INDEX". This index was considered to be an effective qualitative adjunct to chemical quantitation of GAGs in the intervertebral disc.

Alcian Blue↗

Effect of test environment on intervertebral disc hydration.

STUDY DESIGN: Water content of fresh human lumbar intervertebral discs (with adjacent endplates) was assessed in three studies: 1) after each of seven specimen preparation steps. 2) during exposure to either saline spray or a saline bath, and 3) during exposure to a saline bath and 445 N axial compression, either without or with previous exposure to the bath and no compression ("free swelling"). OBJECTIVE: To assess the effect on disc hydration of various aspects of specimen preparation and testing environments. SUMMARY OF BACKGROUND DATA: Water content is an important determinant of disc behavior. Specimen preparation method and testing environments may be important determinants of water content, yet no work appears to have been reported specifically on this topic. METHODS: Endplate-disc-endplate specimens were prepared from refrigerated cadavers within 24 hours of death by transverse sectioning of adjacent vertebral bodies. Water content change was determined by specimen weight change across each time interval of interest. RESULTS: Specimen preparation (including multiple freeze-thaw cycles) produced no water content change. Saline spray and plastic film wrap resulted in no change, but saline bath exposure resulted in a 24% increase over 7 hours, 44% of which occurred in the first 0.5 hour. A subsequent 7 hours of 445 N compression reduced the overall increase to 10%. This was not significantly different from the 8% increase that resulted from initial exposure to saline bath and compression. CONCLUSIONS: Specimen preparation as typically performed and specimen exposure to saline spray and plastic film wrap do not result in hydration change. Exposure to saline bath results in substantial swelling, which can either be reversed or prevented by axial compression in the physiologic range. Whether discs exposed to saline spray and wrap without compression and those exposed to saline bath with compression behave the same and which of these more closely mimics the in vivo condition are important issues for the experimentalist to test.

Adult↗

Secretory leucocyte proteinase inhibitor is produced by human articular cartilage chondrocytes and intervertebral disc fibrochondrocytes.

The objective of this study was to examine the expression of the secretory leucocyte proteinase inhibitor (SLPI) gene by human articular cartilage chondrocytes and intervertebral disc fibrochondrocytes. RNA was extracted from human articular cartilage chondrocytes, synovial fibroblasts and fibrochondrocytes of the annulus fibrosus. Expression was analysed by Northern blotting and hybridisation to a SLPI RNA probe. SLPI was isolated from chondrocyte culture supernatant by gel-permeation and cation-exchange chromatography and examined by a solid-phase enzyme-linked immunofiltration assay for SLPI and by SDS/PAGE using Western blotting with polyclonal IgG samples against bronchial mucus leucocyte proteinase inhibitor and SLPI. This identified biochemically and immunologically the major serine proteinase inhibitory protein synthesised by chondrocytes as SLPI. Secretory leucocyte proteinase inhibitor mRNA was detected in RNA preparations from articular cartilage chondrocytes and annulus fibrosus fibrochondrocytes. The results presented here allowed us to deduce that human articular cartilage chondrocytes and intervertebral disc fibrochondrocytes synthesized SLPI whereas human synovial fibroblasts did not.

Adult↗

[Results of surgical treatment for the intervertebral disc protrusion within thoracic spine].

Results of surgical treatment for intervertebral disc protrusion within thoracic spine in 4 cases are presented. Protrusion had caused spinal compression resulting in neurological impairment (plegia). There were 3 females and 1 male, all in their forties or fifties. Two cases are presented in details, with radiological investigations included. Lateral approach was used at surgery. Neurological deficits subsided completely in all cases. Follow-up ranged from 1-15 years, mean 8 years.

Adult↗

Complete cervical intervertebral disc extrusion with spinal cord injury in the absence of facet dislocation: a case report.

STUDY DESIGN: Complete cervical disc complex extrusion, defined as the extrusion of both cartilaginous end-plates, the entire nucleus pulposus, and portions of the anulus fibrosus, is rare. A case of complete cervical disc complex extrusion with spinal cord injury in the absence of facet dislocation or subluxation in an obtunded patient is reported. OBJECTIVE: To report an unusual presentation of spinal cord injury and the occurrence of complete traumatic cervical disc complex extrusion in the absence of facet dislocation and normal plain radiographic findings. SUMMARY OF BACKGROUND DATA: Traumatic cervical disc herniation occurs in 54% to 80% of patients with facet dislocation. A report of complete extrusion of a cervical intervertebral disc complex (cartilaginous endplate, anulus, and nucleus pulposus) with spinal cord injury in the absence of dislocation has not been described, to the best of the authors' knowledge. METHODS: A clinical and radiographic review of such a case of complete traumatic cervical disc complex herniation in the absence of dislocation was performed. RESULTS: Plain radiographic imaging did not show any injury. A nondisplaced fracture of the left inferior facet joint was evident on computed tomography. The diagnosis of C4-C5 intervertebral disc extrusion was made only after magnetic resonance imaging. The vacuum effect of complete disc extrusion created a "white-out" appearance to the disc space on the sagittal T2 magnetic resonance image. The patient underwent anterior cervical discectomy and fusion with additional posterior cervical fusion. He subsequently regained functional strength against gravity in two of four limbs. He remains completely paraparetic in the left upper extremity and partially paraparetic in the left lower extremity. CONCLUSION: The case report highlights the occurrence of complete traumatic cervical disc extrusion in the absence of facet dislocation with normal plain radiographic findings and consequent spinal cord injury, which can accompany such an injury.

Accidents, Traffic↗

The neuropathology of intervertebral discs removed for low-back pain.

Thirty lower lumbar intervertebral discs (IVDs) removed for low-back pain were examined. There is a profuse non-myelinated axonal network and abundant free nerve terminals in the outer (lateral) half of the annulus fibrosus. The inner annulus and the nucleus pulposus did not contain nerve terminals. No significant changes in the nerve networks could be demonstrated in degenerate IVDs: in particular, ingrowth of nerve terminals into foci of granulomatous tissue was not seen. Early foci of degeneration are clearly shown by Marshall's silver method for metalophil cells. Mucinous filaments are argyrophilic and can be mistaken for axonal structures in the nucleus pulposus.

Back Pain↗