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PubMed2006

Quantitative magnetic resonance imaging of enzymatically induced degradation of the nucleus pulposus of intervertebral discs.

STUDY DESIGN: The structural integrity of the nucleus pulposus (NP) of intervertebral discs was targeted by enzyme-specific degradations to correlate their effects to the magnetic resonance (MR) signal. OBJECTIVE: To develop quantitative MR imaging as an accurate and noninvasive diagnostic tool to better understand and treat disc degeneration. SUMMARY OF BACKGROUND DATA: Quantitative MR analysis has been previously shown to reflect not only the disc matrix composition, but also the structural integrity of the disc matrix. Further work is required to identify the contribution of the structural integrity versus the matrix composition to the MR signal. METHODS: The bovine coccygeal NPs were injected with either enzyme or buffer, incubated at 37 degrees C as static, unloaded and closed 3-disc segments, and analyzed by a 1.5-Tesla MR scanner to measure MR parameters. RESULTS: Collagenase degradation of the NP significantly decreased the relaxation times, slightly decreased the magnetization transfer ratio, and slightly increased the apparent diffusion coefficient. Targeting the proteoglycan and/or hyaluronan integrity by trypsin and hyaluronidase did not significantly affect the MR parameters, except for an increase in the apparent diffusion coefficient of the disc after trypsin treatment. CONCLUSIONS: Our results demonstrate that changes in the structural integrity of matrix proteins can be assessed by quantitative MR.

Animals↗
PubMed1981

Experimental intervertebral disc degeneration: morphologic and proteoglycan changes over time.

An animal model of intervertebral disc degeneration produced by surgical ventral disc herniation in the rabbit is described. Histologic studies showed proliferation of cells on the inner third of the annular wound, with metaplasia into fibrocartilage in the first 2 weeks following injury. Progressive fibrocartilaginous change occurred, reproducing the morphologic changes of disc degeneration over the first 6 weeks involving nearly the entire disc. Proteoglycans (total and newly synthesized) were studied qualitatively and quantitatively for periods of 1 to 200 days after herniation. There were two periods of time during the early course of degeneration when the ability of the proteoglycans to aggregate by interaction with hyaluronic acid was recovered, but this decreases progressively after 6-7 weeks. There was an immediate loss of water content from the injured disc which was restored only transiently during the first 2 days after herniation. Thereafter the water content progressively decreased. The uronic acid content of the disc changed in parallel with the changes in water content. The hyaluronic acid content decreased rapidly after herniation. However, the size of the proteoglycan monomers did not change with degeneration. The biochemical and morphologic changes are correlated, and an early repair mechanism is postulated to exist after injury.

Animals↗
PubMed1996

Herniated lumbar intervertebral discs spontaneously produce matrix metalloproteinases, nitric oxide, interleukin-6, and prostaglandin E2.

STUDY DESIGN: Herniated lumbar disc specimens were obtained from patients undergoing surgical discectomy for persistent radiculopathy and cultured in vitro to determine whether various biochemical agents were being produced. OBJECTIVES: Our hypothesis is that biochemical mediators of inflammation and tissue degradation play a role in intervertebral disc degeneration and in the pathophysiology of radiculopathy. SUMMARY OF BACKGROUND DATA: Low back pain with or without radiculopathy is a significant clinical problem, but the etiology of low back pain and the exact pathophysiology of radiculopathy remain elusive. The biochemical events that occur with intervertebral disc degeneration and, in particular, the role of biochemical mediators of inflammation and tissue degradation have received sparse attention in the literature. There is some preliminary evidence that inflammatory mediators may have an important role in the pathophysiology of radiculopathy. METHODS: Eighteen herniated lumbar discs were obtained from 15 patients undergoing disc surgery. The specimens were cultured and incubated for 72 hours, and the media were collected subsequently for biochemical analysis. Biochemical assays for matrix metalloproteinases, nitric oxide, prostaglandin E2, and a variety of cytokines were performed. As a control group, eight lumbar disc specimens were obtained from four patients undergoing anterior surgery for scoliosis and traumatic burst fractures, and similar biochemical analyses were performed. RESULTS: The culture media from the herniated lumbar discs showed increased levels of matrix metalloproteinase activity compared with the control discs. Similarly, the levels of nitric oxide, prostaglandin E2, and interleukin-6 were significantly higher in the herniated discs compared with the control discs. Interleukin 1 alpha, interleukin-1 beta, tumor necrosis factor-alpha, interleukin-1 receptor antagonist protein, and substance P were not detected in the culture media of either the herniated or control discs. CONCLUSIONS: Herniated lumbar discs were making spontaneously increased amounts of matrix metalloproteinases, nitric oxide, prostaglandin E2, and interleukin-6. These products may be involved intimately in the biochemistry of disc degeneration and the pathophysiology of radiculopathy. Their exact roles certainly need further investigation, but their mere presence implicates biochemical processes in intervertebral disc degeneration.

Adult↗
PubMedDate not supplied

[Intervertebral disc calcification in childhood (author's transl)].

Intervertebral disc calcification in childhood is benign and uncommon. The prognosis is good. Clinically, it is marked by torticollis with neck pain and stiffness. Sometimes, appearances suggestive of sepsis may lead to false diagnosis. The diagnosis is made on good radiological examination which shows opacity of the disc and no other abnormality, though with a tendency to forward protrusion. Healing takes place spontaneously after several days and the calcification disappears after about a year, after a stage of fragmentation. Complications are rare. Treatment should be conservative and symptomatic (braces, analgesics).

Calcinosis↗
PubMed2002

The characterization of versican and its message in human articular cartilage and intervertebral disc.

Splicing variation of the versican message and size heterogeneity of the versican core protein were analyzed in human articular cartilage and intervertebral disc. Splicing variation of the message was studied by PCR analysis to detect the presence or absence of exons 7 and 8, which encode large chondroitin sulfate attachment regions. At all ages in normal cartilage from the third trimester fetus to the mature adult, the presence of the versican isoform possessing exon 8 but not exon 7 (V1) could be readily detected. The message isoforms possessing neither exon 7 nor 8 (V3) or both exons 7 and 8 (V0) were only detectable in the fetus, and the isoform possessing only exon 7 (V2) was never detected. In osteoarthritic cartilage and in adult intervertebral disc the versican message pattern was the same as that observed in the normal adult with only the isoform possessing exon 8 being detected. Core protein heterogeneity was studied by immunoblotting following enzymic removal of the glycosaminoglycan chains from the proteoglycan, using an antibody recognizing the globular G1 region of versican. All articular cartilage extracts from the fetus to the mature adult contained multiple core protein sizes of greater than 200 kDa. The adult cartilage extracts tended to have an increased proportion of the smaller sized core proteins and osteoarthritic cartilage possessed similar core protein sizes to the normal adult. In contrast, intervertebral disc at all post-natal ages showed a greater range of size heterogeneity with a prominent component of about 50 kDa. The abundance of this component increased if the samples were treated with keratanase prior to analysis, suggesting that the G1 region of versican in disc can be substituted with keratan sulfate. The increased presence of versican in the disc relative to articular cartilage may suggest a more pronounced functional role for this proteoglycan, particularly in the nucleus pulposus.

Adolescent↗
PubMed1988

Degradation and biosynthesis of proteoglycans in the nucleus pulposus of canine intervertebral disc after chymopapain treatment.

The effects of chymopapain treatment of the canine intervertebral disc were studied in vivo by monitoring proteoglycan in the nucleus pulposus. Analysis of proteoglycan was carried out by Sepharose CL-4B (Pharmacia Fine Chemicals AB, Stockholm, Sweden) chromatography and electrophoresis. The proteoglycan was degraded to glycosaminoglycans within 1 week after chymopapain treatment. Two weeks later, a proteoglycan smaller than the original appeared in the nucleus pulposus. At 8 weeks after injection, the amount of the newly synthesized proteoglycan, similar in molecular weight to the original, had recovered to about half that of the original, although the new proteoglycan fraction was rich in hyaluronic acid. It was concluded that, following chemonucleolysis with chymopapain, the water-binding capacity of the nucleus pulposus recovered, but that the regenerated nucleus pulposus differed biochemically from the original.

Animals↗
PubMed1978

Anterior intervertebral disc herniations (report of six cases).

Six new patients with anterior intervetebral disc herniations are reported. This is probably one of the most common causes of back pain in children. Excessive sport activity and osteoporosis are probably two main causes of anterior intervertebral disc herniations. Anterior intervertebral disc herniation is a severe complication of prolonged, mild trauma.

Adolescent↗
PubMed1999

Transforming and epidermal growth factors in degenerated intervertebral discs.

We studied the presence of anabolic growth factors in human herniated intervertebral discs (IVD) using a reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. Messenger RNA (mRNA) was isolated from the nucleus pulposus using oligo (dT)25 superparamagnetic beads and probing with gene-specific primers in RT-PCR. mRNA coding for TGF-alpha (3/10), EGF (0/10), TGF-beta1 (0/10) and TGF-beta3 (2/10) or the EGF receptor (EGF-R; 0/10) and TGF-beta type-II receptor (0/10) was found only occasionally. Beta-actin was always present and positive sample controls confirmed the validity of the RT-PCR assay. These RT-PCR findings were confirmed using immunohistochemical staining of EGF and TFG-beta, whereas TGF-alpha protein was always found associated with discocytes. We conclude that the nucleus pulposus of the herniated IVD is vulnerable to proteolytic degradation and depletion of proteoglycans due to the lack and/or low production of anabolic growth factors/receptors which could increase the local synthesis of the extracellular matrix.

Actins↗
PubMed2007

Direct measurement of intervertebral disc maximum shear strain in six degrees of freedom: motions that place disc tissue at risk of injury.

Human intervertebral disc specimens were tested to determine the regions of largest maximum shear strain (MSS) experienced by disc tissues in each of three principal displacements and three rotations, and to identify the physiological rotations and displacements that may place the disc at greatest risk for large tissue strains and injury. Tearing of disc annulus may be initiated by large interlamellar shear strains. Nine human lumbar discs were tagged with radiographic markers on the endplates, disc periphery and with a grid of wires in the mid-transverse plane and subjected to each of the six principal displacements and rotations. Stereo-radiographs were taken in each position and digitized for reconstruction of the three-dimensional position of each marker. Maximum tissue shear strains were calculated from relative marker displacements and normalized by the input displacement or rotation. Lateral shear, compression, and lateral bending were the motions that produced the mean (95% confidence interval) largest mean MSS of 9.6 (0.7)%/mm, 9.0 (0.5)%/mm, and 5.8 (1.6)%/ degrees , respectively, and which occurred in the posterior, posterolateral and lateral peripheral regions of the disc. After taking into account the reported maximum physiological range of motion for each degree of freedom, motions producing the highest physiological MSS were lateral bending (57.8 (16.2)%) and flexion (38.3 (3.3)%), followed by lateral shear (14.4 (1.1)%) and compression (12.6 (0.7)%).

Biomechanical Phenomena↗
PubMed2005

[Evaluation of the intervertebral disc spaces with a low dose radiographic system].

PURPOSE: EOS is a new low dose radiographic system based on micro-pattern gaseous detector technology. The purpose was to compare the value of EOS and MRI in the evaluation of degenerative intervertebral disc disease. METHODS AND MATERIALS: Thirty one fresh lumbar discs were studied with EOS, MRI, and discography. Finally, the discs were dissected for macroscopy. They were graded according to their degenerative level using each imaging modality. Osteophytes, herniations, calcifications, loss of disc height, discographic tears or ligamentous ossification were recorded as anomalies. Morphometric data were measured from the cadaveric sections and low-dose radiographs. RESULTS: Thirty nine percent of the discs were narrowed, with a high correlation between EOS and MRI. Anterior and posterior disc margins were respectively depicted in 22% and 64% of cases with EOS vs 84% and 97% with MRI. MRI showed a disc herniation in 9 cases. The correlation between EOS and MRI was poor for the analysis of anterior and posterior disc margins, and disc herniation. Modic 2 abnormalities were present in 19% of disc levels at MRI. Osteophytes were visible in 65% of disc levels with EOS. The density of the disc space was homogeneous in 65% of cases. In the remaining cases, intradiscal calcifications were detected with EOS and confirmed at macroscopy. CONCLUSION: The EOS low dose images allow a good characterisation of disc abnormalities. Especially, intervertebral disc height, density variations and finer bony details (osteophytes) were detected with high confidence. EOS is a promising new low dose radiographic method for spine imaging.

Aged↗
PubMed1994

Penetration of ceftriaxone into the intervertebral disc.

Concentrations of ceftriaxone in serum and intervertebral disc tissue were determined with high-pressure liquid chromatography in forty-five patients after a single intravenous loading dose of 1000 milligrams given at different intervals before an operation on the spine. The mean serum concentrations in this study corresponded well with reported values. The mean tissue concentrations were 5.6 micrograms per gram (95 per cent confidence interval, 3.6 to 6.8 micrograms per gram) one to less than two hours after administration of the antibiotic, 6.4 micrograms per gram (95 per cent confidence interval, 2.8 to 10.0 micrograms per gram) two to less than four hours, and 3.6 micrograms per gram (95 per cent confidence interval, 0.6 to 6.6 micrograms per gram) at fourteen to less than sixteen hours. These drug concentrations exceed the minimum inhibitory concentration that was effective against 90 per cent of the bacteria for methicillin-sensitive Staphylococcus aureus; for Streptococcus pyogenes, agalactiae, viridans, pneumoniae, and bovis; and for community-acquired Enterobacteriaceae. The average serum-to-tissue ratio was 191:1 at less than one-half hour and 13:1 at less than one and a half hours. The lower values of the 95 per cent confidence intervals for the concentration of the antibiotic exceeded the minimum inhibitory concentrations in the disc tissue against most susceptible bacteria during the period between one and a half and four hours, but a larger bolus would be needed to maintain this level for a longer period (such as in a longer operation) and as prophylaxis against methicillin-sensitive Staphylococcus aureus and coagulase-negative staphylococci.

Body Weight↗
PubMed1975

Genetic factors in the maturation of the canine intervertebral disc.

The composition and structure of the intervertebral discs have been studied in a group of six four-month-old beagles which were full or half-siblings. The animals were raised in identical environment and were comparable weight and size at sacrifice. Collagen, hexosamine, uronic acid, sialic acid and non-collagenous protein analysis (via indirect method) were performed on the nucleus pulposus (NP), transitional zone (TZ) and a annulus fibrosus (AF) of the discs for the lumbo-sacral, lumbar and thoracic spinal levels. Parallel histochemical and morphological examinations of the discs were also performed. The knee joint meniscus, and articular cartilage was also examined for the components described. The results of the analysis for the discs revealed that within this single age group there co-exist two statistically different compositional states. These differences reside largely in the NP and TZ regions and from the analytical values are interpreted as indicating the chondroid and non-chondroid conditions. No such compositional variation was observed for articular cartilage and fibrocartilage (knee joint menisci).

Animals↗
PubMed2004

Disc inflammation potentially promotes axonal regeneration of dorsal root ganglion neurons innervating lumbar intervertebral disc in rats.

STUDY DESIGN: The expression of growth-associated protein 43 (GAP-43), a marker of axonal growth, in the dorsal root ganglion (DRG) neurons innervating the lumbar intervertebral disc was assessed using the retrograde tracing method and immunohistochemistry. OBJECTIVES: To study whether disc inflammation affects GAP-43 expression in DRG neurons innervating the disc in rats. SUMMARY AND BACKGROUND DATA: Persistent inflammation and nerve ingrowth into the inner layer of degenerated discs can be a cause of discogenic pain. Although the presence of GAP-43-expressing nerve fibers in painful discs has been reported, the expression of GAP-43 in DRG neurons innervating the disc has not been studied. METHODS: Seven days after the application of Fluoro-Gold to the L5-L6 disc, 50 microL of saline (n = 10, control group) or complete Freund's adjuvant (n = 10, inflammatory group) was applied to the disc in rats. Ten days after the Fluoro-Gold application, T13-L5 DRGs were double-stained with GAP-43 and either calcitonin gene-related peptide or isolectin B4 (IB4). RESULTS: The percentage of Fluoro-Gold-labeled neurons that were positive for GAP-43 was significantly higher in the inflammatory group (44%) than in the control group (24%, P < 0.001). In both groups, the majority of GAP-43-positive neurons were small and positive for calcitonin gene-related peptide but not IB4. CONCLUSIONS: The present results suggest that disc inflammation potentially promotes axonal growth of DRG neurons innervating the disc. In light of the strong correlation between the expression of calcitonin gene-related peptide and nerve growth factor receptor, it is most likely that nerve growth factor-sensitive DRG neurons extend their axons following disc inflammation.

Animals↗
PubMed2000

The anatomic relation among the nerve roots, intervertebral foramina, and intervertebral discs of the cervical spine.

STUDY DESIGN: An anatomic study of the cervical intervertebral foramina, nerve roots, and intradural rootlets performed using a surgical microscope. OBJECTIVES: To investigate the anatomy of cervical root compression, and to obtain the anatomic findings related to cervical foraminotomy for the treatment of cervical radiculopathy. SUMMARY OF BACKGROUND DATA: Cervical foraminotomy is a procedure performed frequently for the management of cervical radiculopathy. However, anatomic studies of cervical foraminotomy have not been fully elucidated. METHODS: In this study, 18 cadavers were obtained for the study of the cervical spine. All the soft tissues were dissected from the cervical spine. Thereafter, laminectomy and facetectomy were performed on C4 through T1 using a surgical microscope. The nerve roots and surrounding anatomic structures, including intervertebral discs and foramina, were exposed. In addition, the intradural rootlets and their intersegmental connections were observed. RESULTS: The shape of the intervertebral foramina approximated a funnel, the entrance zone being the most narrow part and the root sleeves conical, with their takeoff points from the central dural sac being the largest part. Therefore, compression of the nerve roots occurred at the entrance zone of the intervertebral foramina. Anteriorly, compression of the nerve roots was caused by protruding discs and osteophytes of the uncovertebral region, whereas the superior articular process, the ligamentum flavum, and the periradicular fibrous tissues affected the nerve posteriorly. The C5 nerve roots were found to exit over the middle aspect of the intervertebral disc, whereas the C6 and C7 nerve roots were found to traverse the proximal part of the disc. The C8 nerve roots had little overlap with the C7-T1 disc in the intervertebral foramen. The C6 and C7 rootlets passed two disc levels in the dural sac. Also, a high incidence of the intradural connections between the dorsal rootlets of C5, C6, and C7 segments was found. CONCLUSIONS: This study demonstrated the anatomy of the nerve roots, rootlets, and intervertebral foramina, and may aid in understanding the pathology of cervical radiculopathy. The presence of intradural connections between dorsal nerve roots and the relation between the course of the nerve root and the intervertebral disc may explain the clinical variation of symptoms resulting from-nerve root compression in the cervical spine. To perform cervical foraminotomy for cervical radiculopathy, it is necessary to understand the detailed anatomy of the intervertebral foramina thoroughly.

Aged↗
PubMed1999

Serial changes in the rate of proteoglycan synthesis after chemonucleolysis of rabbit intervertebral discs.

STUDY DESIGN: Serial changes in the rate of proteoglycan synthesis in rabbit discs after chemonucleolytic treatment with chymopapain and chondroitinase ABC were measured using an in vitro method. OBJECTIVES: To determine the retained ability of the intervertebral disc to synthesize proteoglycans after chemonucleolytic treatment. SUMMARY OF BACKGROUND DATA: Most previous studies describe radiologic and histologic changes that occur after chemonucleolytic treatment. However, in humans it is not clear whether reconstitution of the disc space with normal nucleus proteoglycans can occur with time. METHODS: Twenty-five rabbits were treated with chymopapain (10 units/0.1 mL/disc) and chondroitinase ABC (5 units/0.1 mL/disc) by intradisc injection. Five rabbits were killed at each interval, 1, 2, 4, 8 and 12 weeks after injection. Radiologic changes in the disc height were noted, and the rate of proteoglycan synthesis was determined biochemically. RESULTS: After injection, no significant recovery of disc height was seen in either enzyme group after the initial disc narrowing. The average rate of proteoglycan synthesis in control rabbit intervertebral discs, those which had not been surgically treated, was 27.1 (x 10(-6) mmols sulphate/hour/dry weight). Twelve weeks after injection, the values were 21.6 in the saline group, 8.9 in the chondroitinase ABC group, and 8.2 in the chymopapain group. CONCLUSIONS: Doses within the therapeutic range can damage disc cells, at least in the rabbit, so that proteoglycan synthesis declined to 30% of control rates, and no significant recovery of disc height was observed.

Animals↗
PubMedDate not supplied

Modulation of proteoglycan production by cyclic tensile stretch in intervertebral disc cells through a post-translational mechanism.

Proteoglycan production is one of the major extracellular matrix components implicated in the dynamic process of intervertebral disc degeneration. Mechanical stress is an important modulator of the degeneration, but the underlying molecular mechanism at the proteoglycan level remains unclear. The aim of this work was to study the regulation of proteoglycan production by cyclic tensile stretch applied to intervertebral disc annulus fibrosus cells. Matrix metalloproteinases do not seem to be implicated in the regulation of proteoglycan production. By contrast, nitrite oxide production is induced by cyclic tensile stretch, in a time, intensity, and frequency dependant manner. Using a non-specific nitric oxide synthases inhibitor [NG-methyl-L-arginine (L-NMA)], we suppress totally the inhibition of proteoglycan production induced by cyclic tensile stretch suggesting the implication of nitric oxide synthases in the observed phenomenon. Introducing the transcriptional inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole or a more specific inhibitor of nitric oxide synthases II [N-iminoethyl-L-lysine (L-NIL)] did not affect the decreased proteoglycan production, which suggests a post-translational regulation. In contrast, N-omega nitro-L-arginine (L-NNA) a more specific inhibitor of NOS I and III abrogated the cyclic tensile stretch-dependant inhibition of proteoglycan production. These results suggest that cyclic tensile stretch regulates proteoglycan production through a post-translational mechanism involving nitrite oxide. This result could be of interest in the development of local therapeutic strategies aimed at controlling intervertebral disc degeneration.

Animals↗