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PubMed2005

The effects of short-term load duration on anabolic and catabolic gene expression in the rat tail intervertebral disc.

The goal of this study was to determine the time-dependent response of the intervertebral disc cells to in vivo dynamic compression. Forty-seven skeletally mature Wistar rats (>12 months old) were instrumented with an Ilizarov-type device spanning caudal disc 8-9. Using a load magnitude (1 MPa) and frequency (1.0 Hz) that were previously shown to significantly alter mRNA levels in the disc, the effects of 0.5 and 4 h of loading were investigated and compared to a sham group and our previous 2 h results. Annulus and nucleus tissue of loaded (c8-9) and internal control discs (c6-7 and c10-11) were separately analyzed by real-time RT-PCR for levels of mRNA coding for various anabolic (collagen-1A1, collagen-2A1, aggrecan) and catabolic (MMP-3, MMP-13, ADAMTs-4) proteins. In the annulus, mRNA levels increased for Collagen types I & II, and MMP 3 & 13 with increasing load duration. In contrast, the nucleus had the largest increases in aggrecan, ADAMTs-4, MMP-3 and MMP-13 after 2 h of loading, with aggrecan and MMP-13 mRNA levels returning to control values after 4 h of loading. Taken in context with our previous studies, we conclude that intervertebral disc cells from the nucleus and annulus have distinct responses to dynamic mechanical compression in vivo with sensitivity to compression magnitude, frequency and duration.

ADAM Proteins↗
PubMed1983

Changes in the nucleus pulposus of the intervertebral disc in bipedal mice. A light and electron microscopic study.

Bipedal mice were produced by clipping the forelimbs and tails of mice within one week of birth. Using light and electron microscopy, the nucleus pulposus of the lumbar intervertebral disc in the bipedal mice was compared with that in normal mice at three, six, and 12 months of age. In normal neonatal mice, the nucleus pulposus is composed of densely packed notochordal cells, which undergo degenerative changes and decrease in number with age. In the bipedal mice, degenerative changes in the nucleus pulposus were accelerated, and herniation of the nucleus pulposus occurred frequently. At the same time, active chondrocytes associated with cartilage matrix appeared in the nucleus pulposus. This sequence of morphologic changes in the nucleus pulposus of the bipedal mice resembles the age-related changes that occur in the nucleus pulposus of the human intervertebral disc. These morphologic changes can be accelerated by creating abnormal mechanical stress. Chondrocytes in the nucleus pulposus may develop from surrounding cartilaginous tissue--cartilage plates and annulus fibrosus.

Aging↗
PubMed1986

Histological development of intervertebral disc herniation.

Sagittal and horizontal sections of 257 intervertebral discs obtained at autopsy and material obtained from 441 operations for herniation of a disc were examined histologically. In the material that was taken at autopsy, myxomatous degeneration of the annulus fibrosus increased in proportion to the age of the subject. The bundles in the internal layer of the annulus fibrosus reversed their usual direction and showed myxomatous degeneration, sometimes resulting in posterior and anterior convex bulging in the internal layer of the anterior and posterior parts of the annulus fibrosus, respectively. When material from a disc was surgically removed as a single free fragment (as in a complete extrusion or a sequestration type of herniation), annulus fibrosus with myxomatous degeneration was found in most material, while the nucleus pulposus rarely was. These results suggest that, from the standpoint of pathomechanism, a protrusion type of herniation of the annulus fibrosus exists in which only the annulus fibrosus is protruded due to reversal of the bundles of the annulus fibrosus, without involvement of the nucleus pulposus. This type of herniation would be a separate entity from the protrusion type of herniation of the nucleus pulposus that occurs when the nucleus pulposus is protruded through a fissure in the annulus fibrosus.

Adult↗
PubMedDate not supplied

Influence of spine morphology on intervertebral disc loads and stresses in asymptomatic adults: implications for the ideal spine.

BACKGROUND CONTEXT: Sagittal profiles of the spine have been hypothesized to influence spinal coupling and loads on spinal tissues. PURPOSE: To assess the relationship between thoracolumbar spine sagittal morphology and intervertebral disc loads and stresses. STUDY DESIGN: A cross-sectional study evaluating sagittal X-ray geometry and postural loading in asymptomatic men and women. PATIENT SAMPLE: Sixty-seven young and asymptomatic subjects (chiropractic students) formed the study group. OUTCOME MEASURES: Morphological data derived from radiographs (anatomic angles and sagittal balance parameters) and biomechanical parameters (intervertebral disc loads and stresses) derived from a postural loading model. METHODS: An anatomically accurate, sagittal plane, upright posture, quadrilateral element model of the anterior spinal column (C2-S1) was created by digitizing lateral full-spine X-rays of 67 human subjects (51 males, 16 females). Morphological measurements of sagittal curvature and balance were compared with intervertebral disc loads and stresses obtained using a quadrilateral element postural loading model. RESULTS: In this young (mean 26.7, SD 4.8 years), asymptomatic male and female population, the neutral posture spine was characterized by an average thoracic angle (T1-T12) = +43.7 degrees (SD 11.4 degrees ), lumbar angle (T12-S1) = -63.2 degrees (SD 10.0 degrees ), and pelvic angle = +49.4 degrees (SD 9.9 degrees ). Sagittal curvatures exhibited relatively broad frequency distributions, with the pelvic angle showing the least variance and the thoracic angle showing the greatest variance. Sagittal balance parameters, C7-S1 and T1-T12, showed the best average vertical alignment (5.3 mm and -0.04 mm, respectively). Anterior and posterior disc postural loads were balanced at T8-T9 and showed the greatest difference at L5-S1. Disc compressive stresses were greatest in the mid-thoracic region of the spine, whereas shear stresses were highest at L5-S1. Significant linear correlations (p < .001) were found between a number of biomechanical and morphological parameters. Notably, thoracic shear stresses and compressive stresses were correlated to T1-T12 and T4-hip axis (HA) sagittal balance, respectively, but not to sagittal angles. Lumbar shear stresses and body weight (BW) normalized shear loads were correlated with T12-S1 balance, lumbar angle, and sacral angle. BW normalized lumbar compressive loads were correlated with T12-S1 balance and sacral angle. BW normalized lumbar disc shear (compressive) loads increased (decreased) significantly with decreasing lumbar lordosis. Cervical compressive stresses and loads were correlated with all sagittal balance parameters except S1-HA and T12-S1. A neutral spine sagittal model was constructed from the 67 subjects. CONCLUSIONS: The analyses suggest that sagittal spine balance and curvature are important parameters for postural load balance in healthy male and female subjects. Morphological predictors of altered disc load outcomes were sagittal balance parameters in the thoracic spine and anatomic angles in the lumbar spine.

Adult↗
PubMed2002

The internal mechanics of the intervertebral disc under cyclic loading.

The mechanics of the intervertebral disc (IVD) under cyclic loading are investigated via a one-dimensional poroelastic model and experiment. The poroelastic model, based on that of Biot (J. Appl. Phys. 12 (1941) 155; J. Appl. Mech. 23 (1956) 91), includes a power-law relation between porosity and permeability, and a linear relation between the osmotic potential and solidity. The model was fitted to experimental data of the unconfined IVD undergoing 5 cyclic loads of 20 min compression by an applied stress of 1MPa, followed by 40 min expansion. To obtain a good agreement between experiment and theory, the initial elastic deformation of the IVD, possibly associated with the bulging of the IVD into the vertebral bodies or laterally, was removed from the experimental data. Many combinations of the permeability-porosity relationship with the initial osmotic potential (pi(i)) were investigated, and the best-fit parameters for the aggregate modulus (H(A)) and initial permeability (k(i)) were determined. The values of H(A) and k(i) were compared to literature values, and agreed well especially in the context of the adopted high-stress testing regime, and the strain related permeability in the model.

Cadaver↗
PubMed2006

Nucleus pulposus cells express HIF-1 alpha under normoxic culture conditions: a metabolic adaptation to the intervertebral disc microenvironment.

Nucleus pulposus (NP) cells of the intervertebral disc reside in an environment that has a limited vascular supply and generate energy through anaerobic glycolysis. The goal of the present study was to examine the expression and regulation of HIF-1alpha, a transcription factor that regulates oxidative metabolism in nucleus pulposus cells. Nucleus pulposus cells were isolated from rat, human, and sheep disc and maintained at either 21% or 2% oxygen for various time periods. Cells were also treated with desferrioxamine (Dfx), a compound that mimics the effects of hypoxia (Hx). Expression and function of HIF-1alpha were assessed by immunofluorescence microscopy, Western blot analysis, gel shift assays, and luciferase reporter assays. In normoxia (Nx), rat, sheep, and human nucleus pulposus cells consistently expressed the HIF-1alpha subunit. Unlike other skeletal cells, when maintained under low oxygen tension, the nucleus pulposus cells exhibited a minimal induction in HIF-1alpha protein levels. Electromobility shift assays confirmed the functional binding of normoxic HIF-1alpha protein to its putative DNA binding motif. A dual luciferase reporter assay showed increased HIF-1alpha transcriptional activity under hypoxia compared to normoxic level, although this induction was small when compared to HeLa and other cell types. These results indicate that normoxic stabilization of HIF-1alpha is a metabolic adaptation of nucleus pulposus cells to a unique oxygen-limited microenvironment. The study confirmed that HIF-1alpha can be used as a phenotypic marker of nucleus pulposus cells.

Adaptation, Physiological↗
PubMed1985

Some additional suggestions for an intervertebral disc prosthesis.

The paper, which refers to previous original papers on the subject, gives suggestions indicating four intervertebral disc prostheses, one utilizing bone cement for fixation to the bony substance of the motion segment. In the paper a modification of this inter-vertebral disc prosthesis offers a possibility for fixation of the prosthesis by osteointegration.

Biomechanical Phenomena↗
PubMed1989

Hierarchical structure of the intervertebral disc.

Optical microscope techniques are used to characterize the hierarchical structure of the collagenous components of the human intervertebral disc. In the anterior annulus fibrosus, the thickness of lamellae increases abruptly 2 mm inward from the edge of the disc, dividing the annulus into peripheral and transitional regions. Lamellae in the lateral and posterior aspects of the disc have a broad distribution of lamellar thicknesses throughout the annulus. In alternating lamellae, fibers are inclined with respect to the vertical axis of the spine in a layup structure. From the edge of the disc inward to the nucleus, this interlamellar angle decreases from +62 to +45 degrees. Within lamellae, the collagen fibers exhibit a planar crimped morphology. The plane of the waveform is inclined with respect to the vertical axis by the interlamellar angle. From the edge of the disc inward, the crimp angle increases from 20 to 45 degrees and the crimp period decreases from 26 to 20 um. A hierarchical model of the intervertebral disc has been developed that incorporates these morphological gradients.

Adult↗
PubMed1990

Growth related increase in rat intervertebral disc size: a quantitative radiographic and histologic comparison.

Methods were developed for measuring the width of lumbar intervertebral discs in Sprague-Dawley rats. Rates of growth of the L4-5 and L5-6 discs in a rostral-caudal direction in an approximately midsagittal plane were reported from analysis of radiographic and histologic data. The radiographic data correlated with histologic data (p less than 0.001). It was concluded that the L4-5 and L5-6 intervertebral discs continue to increase in size throughout the first 18 months of life of the rat, well past the age of rapid gain in body weight.

Animals↗
PubMed1992

Non-linear finite element analysis of formation and treatment of intervertebral disc herniae.

A biomechanical model of the spine motion segment L2/L3 consisting of the truncated vertebrae, endplates, intervertebral disc and pieces of anterior and posterior longitudinal ligaments has been used for a computer simulation study. A non-linear finite element analysis has shown that small loads compressing the spine, not greater than those occurring in everyday life, cause loss of stability of an intervertebral disc, resulting in lateral dislocation of its nucleus pulposus. This could be a potential cause of discopathy. The model indicates that conservative therapy of herniated disc by the traction method may result in retraction of hernia by about 40 per cent.

Biomechanical Phenomena↗
PubMed1994

Substance P in intervertebral discs. Binding sites on vascular endothelium of the human annulus fibrosus.

The annulus fibrosus of the human intervertebral disc is sparsely innervated, some of the fibers containing substance P. We could demonstrate, by autoradiography, binding sites for substance P localized on the endothelium of small blood vessels in the annulus fibrosus of human intervertebral discs removed during anterior fusion for back pain. In binding inhibition studies, binding of 125I-Bolton Hunter-substance P was inhibited by unlabeled substance P and the related tachykinins neurokinin A and neurokinin B with a rank order of potency substance P > NKA > NKB. Specific binding was reduced > 75 percent by 5'-guanylylimidodiphosphate, indicating G-protein coupling. These features are characteristic of an NK1 receptor through which vascular effects, i.e., vasodilation, plasma extravasation and angiogenesis of substance P, are mediated. The presence of NK1 receptors on blood vessels in the annulus fibrosus may indicate a role for substance P in tissue repair although acute proinflammatory effects may contribute to discogenic pain.

Adult↗
PubMed1998

Keratan sulfate as a potential biomarker of loading of the intervertebral disc.

STUDY DESIGN: A review of the literature. OBJECTIVE: To investigate the potential of serum levels of keratan sulfate as a biomarker of the effects of loading of the spine. SUMMARY OF BACKGROUND DATA: Exposure to mechanical loading of the spine causes changes in metabolism of intervertebral discs, eventually leading to accelerated disc degeneration. This process is characterized by the degradation of proteoglycans, which is reflected by an increase in the blood level of proteoglycan components. The serum level of keratan sulfate, an epitope present on these proteoglycan components, has been suggested as a marker of changes in metabolism of cartilaginous tissues. METHODS: A review of the literature on serum keratan sulfate levels in relation to degenerative changes in cartilaginous tissue. RESULTS: In a number of studies keratan sulfate in serum was reported to be related to degeneration of articular cartilage in patients with osteoarthritis. In addition, massive and rapid degradation of intervertebral discs was determined to result in a large rise in serum keratan sulfate levels. Whether degenerative changes of intervertebral discs induced by mechanical stress also cause a detectable increase in serum keratan sulfate should be subjected to further investigation. CONCLUSION: Quantification of keratan sulfate in serum offers a promising measure for the early effects of mechanical loading of the spine, but research is needed for validation.

Biomarkers↗
PubMed2005

[Study on the effect of combined therapy of Chinese and Western medicines and gene therapy (GT) on cartilage end-plate calcification in rabbits with cervical intervertebral disc regression].

OBJECTIVE: To study the relationship between cartilage end-plate calcification (CEC) and cervical intervertebral discs regression (CIDR) in rabbits, and to study the inhibitory effect of combined therapy of Kanggu Zengsheng Capsule (KZC) ansforming growth factor-apsule (TGF-PLC) and igene therapy (GT) on CEC by measuring the thickness of CEC layer. METHODS: Thirty-five New Zeland rabbits of 4 months old were selected to establish cervical dynamic imbalance rabbit model for inducing CIDR (No disposal was given to rabbits in the normal control group). Seven months after operation, combined therapy of KZC and PLC were given, in doses calculated by body weight, to the modeled rabbits in the drug treated group with CEC of either superficial layer or full layer, twice a dantly by gavage for 30 successive days. While to those in the gene therapy group, the recombinant plasmid DNA with transforming growth factor-beta1 (TGF-beta1) was injected once their intervertebral discs (ID) of C(2-3) C(3-4) and C(4-5), 20 microl for each injection. One month later, all rabbits were sacrificed with periotic venous gas embolic method and their ID of C(4-5) (including partial body of the upper and lower vertebrae) was resected. The degree of CIDR was evaluated morphologically, and the thickness of CEC in rabbits was measured and compared between groups. RESULTS: Thickness of CEC in the model group, either of superficial layer or of full layer, was significantly more than that in the normal control group with significant difference. Both combined KZC and PLC therapy and gene therapy showed significant inhibitory effects on CEC in treating CIDR (P < 0.05). CONCLUSION: CEC is the initial factor of CIDR with highly positive correlation. Both combined therapy of KZC and PLC and gene therapy can significantly inhibit CEC.

Animals↗
PubMed1981

Chemonucleolysis for relief of sciatica due to a herniated intervertebral disc.

Chemonucleolysis is the nonoperative chemical removal of displaced lumbar disc material. The enzyme chymopapain, which has a wide margin of safety between its effective therapeutic and toxic doses, is effective in the management of sciatica due to a herniated intervertebral disc. The patient will have leg pain as the dominant symptom and a 50% reduction in straight-leg raising with or without bowstring discomfort and crossover pain. Neurologic symptoms and signs are usual, as are abnormal results of contrast studies, which will verify the level of involvement. In 220 randomly selected patients who met criteria for the diagnosis of sciatica due to a herniated intervertebral disc and did not have psychogenic or nonorganic spinal pain, a spinal stenosis or a history of a previous, unsuccessful operation to relieve the sciatica, chemonucleolysis had a success rate of 80%. The only complications were a severe anaphylactic reaction in two patients and lesser, delayed reactions in five others. All of the reactions were successfully treated. Of the 45 patients in whom chemonucleolysis was unsuccessful, 38 underwent a laminectomy. In 3 of the 38 the results of chemonucleolysis were initially good, but later the disc herniation recurred; thus, the long-term treatment failure rate was 1.4%.

Adult↗
PubMed2003

Bone morphogenetic protein-2 facilitates expression of chondrogenic, not osteogenic, phenotype of human intervertebral disc cells.

STUDY DESIGN: In vitro experiment using bone morphogenetic protein-2 (BMP-2) and human intervertebral disc (IVD) cells. OBJECTIVES: To demonstrate the effect of BMP-2 on mRNAs expression (collagen type I, collagen type II, aggrecan, and osteocalcin), proteoglycan synthesis, expression of alkaline phosphatase, bone nodule formation in human IVD cells. SUMMARY OF BACKGROUND DATA: BMP-2 was widely known as a powerful agent for osteoinduction and a crucial growth factor for early chondrogenesis and maintenance of cartilaginous phenotype. BMP-2 proved to be effective in stimulating proteoglycan synthesis in articular chondrocytes and IVD cells. Nevertheless, the effect of BMP-2 on IVD cells, whether chondrogenic or osteogenic, was not thoroughly elucidated in transcriptional level and histochemical stains. MATERIALS AND METHODS: Human IVDs were harvested and enzymatically digested. Then IVD cells were cultured three-dimensionally in alginate beads. Osteoblasts were cultured from cancellous bone of ilium for histochemical stains. Recombinant human BMP-2 (rhBMP-2) was produced by Chinese hamster ovary cells after transduction of BMP-2 cDNA, then concentrated and purified. Then IVD cell cultures were exposed to various concentrations of rhBMP-2. Reverse transcription-polymerase chain reaction for mRNA expression of aggrecan, collagen type I, collagen type II, and osteocalcin was performed. Newly synthesized proteoglycan was measured by 35S-sulfate incorporation on Sephadex G-25 M in PD 10 columns. As a histochemical examination, alkaline phosphatase and Alizarin red-S stains were used to detect osteogenic marker and bone nodule formation, respectively. RESULTS: In the rhBMP-2 treated cultures, there was increased newly synthesized proteoglycan (67% in 300 ng/mL and 200% in 1,500 ng/mL of rhBMP-2) and up-regulated expression of aggrecan, collagen type I, and collagen type II mRNA over untreated control. However, rhBMP-2 did not up-regulate expression of osteocalcin mRNA in the given dose and culture period. IVD cell cultures with rhBMP-2 showed no evidence of bone formation in histochemical stains, i.e., alkaline phosphatase and Alizarin red-S, while osteoblast culture exhibited strong positive stains. CONCLUSIONS: The rhBMP-2 clearly up-regulated mRNA expression of chondrogenic components and also stimulated proteoglycan synthesis without expression of osteogenic phenotype. Taken together, this study raise the possibility of rhBMP-2 can be anabolic agent for regenerating matrix of intervertebral disc.

Adult↗