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PubMed2005

Segmental variation in the in vitro cell metabolism of nucleus pulposus cells isolated from a series of bovine caudal intervertebral discs.

STUDY DESIGN: This study focuses on the association between cell metabolism and molecular matrix composition of nucleus pulposus (NP) tissue with spine level in sequential bovine caudal intervertebral discs. OBJECTIVE: To explore the hypothesis that the molecular composition of NP tissue and corresponding cell metabolism varies with caudal spine. A secondary hypothesis is tested that potential cellular differences are maintained after monolayer culture. SUMMARY OF BACKGROUND DATA: In articular cartilage, cell metabolism and molecular matrix composition are influenced by loading history. This may also be a feature of intervertebral discs in series. METHODS: NP cells (nonpooled or level pooled) were isolated from four sequential bovine caudal intervertebral discs (levels 3-4, 4-5, 5-6, and 6-7) and cultured in alginate beads immediately or following monolayer culture. Levels of 3H-TdR (proliferation) and 35SO4 (GAG synthesis) incorporation were determined from 14 animals. In a separate set of 6 animals, total content of water, DNA, collagen (and type), GAG (and type) were also determined. RESULTS: The rate of 3H-TdR and 35SO4 incorporation in freshly isolated NP cells increased nonlinearly from level 3-4 to 6-7 (P < 0.05). Monolayer cultured cells retained level-specific differences for 35SO4 and 3H-TdR incorporation similar to that of freshly isolated cells. GAG content and chondroitin sulfate proportion decreased distally (P < 0.05); however, total collagen and Type I proportion increased distally (P < 0.05). No significant differences in water or DNA content could be determined. CONCLUSIONS: The results support the hypothesis that level-specific differences in NP cell metabolism and molecular composition are dependent on spine level potentially reflecting subtle mechanical differences between levels. Retention of level-specific differences in monolayer may suggest a certain level of cell "programming." This may be important for cellular strategies to repairspecific sites of degeneration.

Animals↗

PubMed1995

The effect of prophylactic antibiotics on iatrogenic intervertebral disc infections. a rabbit model.

STUDY DESIGN: A rabbit model was used to test the efficacy of two commonly used prophylactic antibiotics, cefazolin and vancomycin, in preventing iatrogenically introduced Staphylococcus aureus intervertebral disc infections. OBJECTIVE: This study was performed to assess the efficacy of two prophylactic antibiotics in preventing iatrogenically introduced Staphylococcus aureus intervertebral disc infections. SUMMARY OF BACKGROUND DATA: Previous studies have had conflicting results regarding the penetration of antibiotics into the nucleus pulposus and their ability to eradicate infection. METHODS: In this study, 40 adult New Zealand White rabbits underwent inoculation of 10(1) or 10(3) Staphylococcus aureus/ml into 3-6 lumbar intervertebral discs under direct visualization. Either no antibiotics (control groups) or various preoperative and postoperative dosing schedules of cefazolin or vancomycin were given intravenously. Five days after surgery, the discs were harvested and cultured. RESULTS: All 40 discs inoculated in the control groups became infected. None of the 35 discs inoculated in the cefazolin groups became infected. Infection developed in 23 of 107 discs inoculated in the vancomycin groups. Most notable of these were 17 of 17 positive cultures in animals given vancomycin 8 hours preoperatively only. CONCLUSIONS: Based on these results, it was concluded that intravenous cefazolin or vancomycin given within 1 hour before surgery can effectively prevent postoperative discitis. No advantage was found with additional postoperative antibiotics.

Animals↗

PubMed2003

Effects of immobilization and dynamic compression on intervertebral disc cell gene expression in vivo.

STUDY DESIGN: An in vivo analysis of the intervertebral disc's cellular response to dynamic compression and immobilization was performed using a rat-tail model. OBJECTIVE: To assess the effects of immobilization and short-term dynamic compression on intervertebral disc cell expression of anabolic and catabolic genes. SUMMARY OF BACKGROUND DATA: Static compressive loads applied in vivo alter the composition of the disc matrix and cell viability in a dose-dependent manner. The effects of in vivo dynamic compression, which is a more physiologic load, and reported risk factor for low back pain have not been investigated. METHODS: An Ilizarov-type device was implanted on the rat tail and used to determine the effects from 72 hours of immobilization (n = 6), 2 hours of dynamic compression (1 MPa/0.2 Hz) (n = 8), and the coupled effect of immobilization followed by compression (n = 8). Real-time reverse transcription-polymerase chain reaction was used to measure changes in anabolic and catabolic gene levels relative to both internal control subjects and a sham-operated group (n = 7). RESULTS: Immobilization and dynamic compression affect anabolic and catabolic genes, with an overall downregulation of types 1 and 2 collagen and upregulation of aggrecanase, collagenase, and stromelysin in the anulus. The effects of immobilization and compression appear to be additive for collagen types 1 and 2 in the anulus, but not in the nucleus, and not for catabolic genes. CONCLUSIONS: Short-duration dynamic compression and immobilization alter gene expression in the rat disc. In studying the response of the disc to loading, it is necessary to look at both anabolic and catabolic pathways, and to consider strain history.

Aggrecans↗

PubMedDate not supplied

In situ calibration of an oxygen sensor in intervertebral discs.

Intradiscal hypoxia has been suspected of playing a role in the pathogenesis of degenerative back diseases. Investigating this hypothesis requires quantitative methods for measuring PO2 in the intervertebral disc. Tissue oxygen tensions are usually measured with a membrane-covered oxygen-consuming sensor. Calibrating this sensor is complicated by the existence of an oxygen-diffusion gradient in front of the membrane. This article describes a method based on mass spectrometry to measure the oxygen tension in the intervertebral disc and an 'in situ' calibration technique, which is able to compensate for errors caused by the oxygen-diffusion gradient. The principle of the calibration technique is to measure the signal of an inert reference gas, whose partial pressure in the intervertebral disc is known, and to utilize the measured signal to estimate the magnitude of the oxygen-diffusion gradient.

Humans↗

PubMed2006

Outer annulus tears have less effect than endplate fracture on stress distributions inside intervertebral discs: relevance to disc degeneration.

BACKGROUND: Annulus tears and endplate fracture are common lesions in human intervertebral discs. Both cause degenerative changes in animal models, but the time course appears to be different. The purpose of the present experiment is to compare the effects of outer annulus tears and endplate fracture on intradiscal stresses. We hypothesise that endplate fracture provides a greater stimulus for disc degeneration. METHODS: Seven cadaveric lumbar "motion segments" aged 49-70 years were compressed at 2 kN while the distribution of compressive stress was measured in each disc by pulling a 1.3 mm-diameter pressure transducer along its mid-sagittal diameter. Measurements were repeated after rim tears were simulated by 10 mm-deep scalpel cuts into the outer anterior annulus. The first cut was horizontal, 15 mm to the right of the disc midline, near the junction with the upper endplate. The second cut was vertical, 15 mm to the left of the disc midline, at mid-disc height. The third cut was horizontal, in the disc midline and at mid-disc height, so that the cut passed through the needle hole of the pressure transducer. Stress profiles were recorded in three postures and at two load levels, after each cut. Stress measurements were repeated a final time following compressive overload sufficient to fracture the endplate. FINDINGS: Outer annulus tears had negligible effect on compressive stress distributions in the annulus fibrosus more than 15 mm from the scalpel cut, and they caused nucleus pressure to fall by only 1% (SD 1.3%, NS). In contrast, endplate fracture reduced nucleus pressure by 37% (P=0.004) and increased maximum stress within the posterior annulus by 93% (P=0.033). INTERPRETATION: Outer annulus tears have less (immediate) effects on intradiscal compressive stresses than endplate fracture, supporting our hypothesis.

Aged↗

PubMed1988

Pain descriptions and surgical findings in patients with herniated lumbar intervertebral discs.

The diagnostic value of pain mapping in patients with lumbago sciatica and herniated intervertebral discs is evaluated, somatosensory pain descriptions being compared with surgical findings. Information has been extracted from complex data sets using a multivariate data analysis program (FCVPC-87). Superficially experienced pain may represent pronounced nerve root affliction. Pain relief in particular body positions was related to extruded discs and additional osseous stenosis, and rarely occurring relief to derangement of nerve root circulation. Neither somatosensory pain quality descriptions nor pain intensity variables were found to be of diagnostic value. Long duration of symptoms was found together with nerve root atrophy. Affliction of the L5 and S1 nerve roots showed differences with regard to pain localization and intensity and some surgical findings. Pain mapping may represent an indication of the patho-anatomical substratum in patients with herniated lumbar intervertebral discs.

Adolescent↗

PubMed2006

Frequency response of pig intervertebral disc cells subjected to dynamic hydrostatic pressure.

The pathogenesis of vibration-induced disorders of intervertebral disc at the cellular level is largely unknown. Dynamic loads with frequencies close to that of the in vivo human spine resonant frequency (4-6 Hz) have a destructive effect, which may induce extracellular disc matrix (ECM) degradation. To investigate this issue, three-dimensional (3D) alginate cultures of normal pig intervertebral disc nucleus and inner annulus cells were tested under dynamic hydrostatic loading. Alginate cultures of each region were divided into six groups; five groups were exposed to cyclic hydrostatic pressures of frequencies 1, 3, 5, 8, and 10 Hz with the same amplitude (1 MPa), and group 6 was the control group (no loading). Cultures of different groups were loaded for 3 days (30 min daily) in a hydraulic chamber. Effects of loading frequency on disc collagen and protein metabolism were investigated by measuring 3H-proline-labeled proteins associated with the cells in the extracellular matrix and release of 3H-proline-labeled molecules into culture medium. The results indicated a poor synthesis rate and more degradation near the 5 Hz frequency. The repeatability of experiments was verified by performing two experiments with the same protocol. Both experiments indicated that a threshold frequency of around 5 Hz disrupted protein metabolism.

Animals↗

PubMed2000

Mechanical initiation of intervertebral disc degeneration.

STUDY DESIGN: Mechanical testing of cadaveric lumbar motion segments. OBJECTIVES: To test the hypothesis that minor damage to a vertebral body can lead to progressive disruption of the adjacent intervertebral disc. SUMMARY OF BACKGROUND DATA: Disc degeneration involves gross structural disruption as well as cell-mediated changes in matrix composition, but there is little evidence concerning which comes first. Comparatively minor damage to a vertebral body is known to decompress the adjacent discs, and this may adversely affect both structure and cell function in the disc. METHODS: In this study, 38 cadaveric lumbar motion segments (mean age, 51 years) were subjected to complex mechanical loading to simulate typical activities in vivo while the distribution of compressive stress in the disc matrix was measured using a pressure transducer mounted in a needle 1.3 mm in diameter. "Stress profiles" were repeated after a controlled compressive overload injury had reduced motion segment height by approximately 1%. Moderate repetitive loading, appropriate for the simulation of light manual labor, then was applied to the damaged specimens for approximately 4 hours, and stress profilometry was repeated a third time. Discs then were sectioned and photographed. RESULTS: Endplate damage reduced pressure in the adjacent nucleus pulposus by 25% +/- 27% and generated peaks of compressive stress in the anulus, usually posteriorly to the nucleus. Discs 50 to 70 years of age were affected the most. Repetitive loading further decompressed the nucleus and intensified stress concentrations in the anulus, especially in simulated lordotic postures. Sagittal plane sections of 15 of the discs showed an inwardly collapsing anulus in 9 discs, extreme outward bulging of the anulus in 11 discs, and complete radial fissures in 2 discs, 1 of which allowed posterior migration of nucleus pulposus. Comparisons with the results from tissue culture experiments indicated that the observed changes in matrix compressive stress would inhibit disc cell metabolism throughout the disc, and could lead to progressive deterioration of the matrix. CONCLUSIONS: Minor damage to a vertebral body endplate leads to progressive structural changes in the adjacent intervertebral discs.

Adult↗

PubMed1975

Factors involved in the nutrition of the human lumbar intervertebral disc: cellularity and diffusion of glucose in vitro.

Post-mortem specimens of the human lumbar (L4-L5) intervertebral disc have been studied histologically and physico-chemically. Blood vessels were found only at the margin of the anulus fibrosus and in the vertebral marrow spaces. Contact between disc tissue and marrow spaces occupied about 10% of the bone-cartilage interface. The disc was most cellular at the periphery of the anulus fibrosus and in the hyaline cartilage next to the vertebral bone. Cellularity declined towards the nucleus pulposus where it achieved a low constant value. The cell density of the disc as a whole was about 60000 cells/mm3. For glucose, the diffusion coefficient of the anulus fibrosus and hyaline cartilage end plate was 2.5 cm2/sec and 2.4 cm2/sec respectively at 37 degrees C, comparable to that of cartilage elsewhere. The permeability of the bone-cartilage interface was low, particularly in the peripheral part. Calculations, based on the present findings and derived values for glucose utilization in disc tissue, indicate that nutritional conditions in the intervertebral disc are more critical than, for example, in articular cartilage.

Adolescent↗

PubMed1950

Ruptured cervical intervertebral discs.

Intractable pain in the neck and upper extremity is frequently radicular in character and mechanical in origin. Rupture of a cervical intervertebral disc is the most common cause. Diagnosis and localization are more difficult in the cervical than in the lumbar region. Pantopaque myelogram should be considered only as a mean of localization. Operation should be preceded by a thorough trial of conservative treatment.

Cervical Vertebrae↗

PubMed2002

Cytokines and growth factors in the protruded intervertebral disc of the lumbar spine.

Nerve root irritation induced by factors produced by the intervertebral disc may play a crucial role in the pathophysiology of sciatic pain production. In this study we used immunohistochemistry to investigate the presence of transforming growth factor-beta1 (TGF-beta1), insulin-like growth factor-1 (IGF-1), interleukin-6 (IL-6), IL-6-receptor (IL-6R) and fibronectin in lumbar disc bioptic specimens from 30 patients with disc herniation (protrusion type). Chondrocytes of herniated discs stained positive for TGF-beta1, IGF-1, IL-6 and fibronectin. We demonstrated for the first time the presence of IL-6-R in the chondrocytes of herniated tissue. Specimens from autoptic healthy tissue were used as controls. In these sections no immunoreaction for TGF-beta1, IL-6, or IL-6R was found, while they expressed IGF-1 and fibronectin, but in lower quantities than herniated discs. These results demonstrated the production of factors such as TGF-beta1, IGF-1, IL-6, IL-6R and fibronectin at the site of lumbar disc herniation.

Adult↗

PubMed1958