In vitro torsion-induced stress distribution changes in porcine intervertebral discs.

STUDY DESIGN: A cadaveric porcine spine motion segment experiment was conducted. OBJECTIVE: To test the hypothesis that small vertebral rotations cause increased stress in the anulus while decreasing stress in the nucleus through stiffening of the anulus. SUMMARY OF BACKGROUND DATA: Stress profiles of the intervertebral disc reportedly depend on degeneration grade and external loading. Increased stress in the anulus was found during asymmetric loading. In addition, depressurization of the nucleus combined with an instantaneous disc height increase was found when small (<2 degrees ) axial vertebral rotations were applied. METHODS: Seven lumbar porcine cadaveric motion segments consisting of two vertebrae and the intervening disc with ligaments were loaded in the neutral position with 340 N of compression. Stress profiles were obtained in the neutral position, then after 0.5 degrees and 1 degrees axial rotation of the bottom vertebral body. The distribution of compressive stress in the disc matrix was measured by pulling a miniature pressure transducer through the disc along a straight path in the midfrontal plane. Stress profiles were measured in vertical (0 degrees ) and horizontal (90 degrees ) orientation. RESULTS: Deformation of the anulus by small axial rotations of the lower vertebra instantaneously decreased the horizontally and vertically measured stress in the nucleus while increasing stress in the anulus. A 1-hour period of creep loading decreased the stresses in the nucleus and the anulus 20% to 30%, depending on the orientation, but the effect of an increasing stress in the anular region after axial rotation persisted. CONCLUSIONS: The compressive Young's modulus of the composite anulus tissue increases instantaneously when small axial rotations are applied to porcine spine motion segments. This is accompanied by decreased stress in the nucleus pulposus, increased stress in the anulus fibrosus, changes in the stress profile superimposed on and independent of prolonged viscoelastic creep and dehydration, and changes in stress distribution independent of horizontal and vertical orientation.     

Stress in lumbar intervertebral discs during distraction: a cadaveric study

Background contextThe intervertebral disc is a common source of low back pain (LBP). Prospective studies suggest that treatments that intermittently distract the disc might be beneficial for chronic LBP. Although the potential exists for distraction therapies to affect the disc biomechanically, their effect on intradiscal stress is debated.PurposeTo determine if distraction alone, distraction combined with flexion, or distraction combined with extension can reduce nucleus pulposus pressure and posterior annulus compressive stress in cadaveric lumbar discs compared with simulated standing or lying.Study designLaboratory study using single cadaveric motion segments.Outcome measuresStrain gauge measures of nucleus pulposus pressure and compressive stress in the anterior and posterior annulus fibrosus.MethodsIntradiscal stress profilometry was performed on 15 motion segments during 5 simulated conditions: standing, lying, and 3 distracted conditions. Disc degeneration was graded by inspection from 1 (normal) to 4 (severe degeneration).ResultsAll distraction conditions markedly reduced nucleus pressure compared with either simulated standing or lying. There was no difference between distraction with flexion and distraction with extension in regard to posterior annulus compressive stress. Discs with little or no degeneration appeared to distribute compressive stress differently than those with moderate or severe degeneration.ConclusionsDistraction appears to predictably reduce nucleus pulposus pressure. The effect of distraction therapy on the distribution of compressive stress may be dependent in part on the health of the disc.     

Peak stresses observed in the posterior lateral anulus

STUDY DESIGN: The stress distributions within cadaveric lumbar intervertebral discs were measured for a range of loading conditions. OBJECTIVES: To examine the distribution of stress across the area of the intervertebral disc and to compare regional variations in peak stress during compression loading with various flexion angles. SUMMARY OF BACKGROUND DATA: The rate of disc degeneration and the occurrence of low back disorders increase with higher mechanical loading of the spine. The largest peak stresses occur in the anulus. METHODS: Human lumbar L2--L3 and L4--L5 cadaver functional spinal units were obtained and tested. The distribution of disc stress was measured using a pressure probe with loads applied, pure compression and compression with 5 degrees of either flexion or extension. RESULTS: Stress profiles were recorded across the intervertebral disc at a compressive force of 1000 N and each of the three flexion-extension angles. The highest values (2.99 +/- 1.31 MPa) were measured during extension-compression lateral to the midline of the disc in the posterior anulus. The pressure in the nucleus was relatively unchanged by flexion angle remaining about 1.00 MPa for a 1000-N compression. CONCLUSIONS: Pressure measurements of the cadaveric nucleus have been used to validate models of lumbar spine loading and to evaluate the risk of low back injury and disc herniation. Previous observations limited to midsagittal measurements of the nucleus did not identify the regions of highest stress. The highest values observed here within the posterolateral anulus correspond to common sites of disc degeneration and herniation.     

Intervertebral disc degeneration can predispose to anterior vertebral fractures in the thoracolumbar spine.

Mechanical experiments on cadaveric thoracolumbar spine specimens showed that intervertebral disc degeneration was associated with reduced loading of the anterior vertebral body in upright postures. Reduced load bearing corresponded to locally reduced BMD and inferior trabecular architecture as measured by histomorphometry. Flexed postures concentrated loading on the weakened anterior vertebral body, leading to compressive failure at reduced load. INTRODUCTION: Osteoporotic fractures are usually attributed to age-related hormonal changes and inactivity. However, why should the anterior vertebral body be affected so often? We hypothesized that degenerative changes in the adjacent intervertebral discs can alter load bearing by the anterior vertebral body in a manner that makes it vulnerable to fracture. MATERIALS AND METHODS: Forty-one thoracolumbar spine "motion segments" (two vertebrae and the intervertebral disc) were obtained from cadavers 62-94 years of age. Specimens were loaded to simulate upright standing and flexed postures. A pressure transducer was used to measure the distribution of compressive "stress" inside the disc, and stress data were used to calculate how compressive loading was distributed between the anterior and posterior halves of the vertebral body and the neural arch. The compressive strength of each specimen was measured in flexed posture. Regional volumetric BMD and histomorphometric parameters were measured. RESULTS: In the upright posture, compressive load bearing by the neural arch increased with disc degeneration, averaging 63 +/- 22% (SD) of applied load in specimens with severely degenerated discs. In these specimens, the anterior half of the vertebral body resisted only 10 +/- 8%. The anterior third of the vertebral body had a 20% lower trabecular volume fraction, 16% fewer trabeculae, and 28% greater intertrabecular spacing compared with the posterior third (p < 0.001). In the flexed posture, flexion transferred 53-59% of compressive load bearing to the anterior half of the vertebral body, regardless of disc degeneration. Compressive strength measured in this posture was proportional to BMD in the anterior vertebral body (r2 = 0.51, p < 0.001) and inversely proportional to neural arch load bearing in the upright posture (r2 = 0.28, p < 0.001). CONCLUSIONS: Disc degeneration transfers compressive load bearing from the anterior vertebral body to the neural arch in upright postures, reducing BMD and trabecular architecture anteriorly. This predisposes to anterior fracture when the spine is flexed.     

Gradual disc prolapse

Fifty-two cadaveric lumbar motion segments were subjected to fatigue loading in compression and bending to determine if the intervertebral discs could prolapse in a gradual manner. Prior to testing, the nucleus pulposus of each disc was stained with a small quantity of blue dye and radiopaque solution. This enabled the progress of any gradual prolapse to be monitored by direct observation and by discogram. Six discs developed a gradual prolapse during the testing period. The injury starts with the lamellae of the annulus being distorted to form radial fissures and then nuclear pulp is extruded from the disc and leaks into the spinal canal. Discs most commonly affected were from the lower lumbar spine of young cadavers. Tests on ten older discs with pre-existing ruptures showed that such discs are stable and do not leak nuclear pulp.     

Intradiscal solid phase displacement as a determinant of the centripetal fluid shift in the loaded intervertebral disc

STUDY DESIGN: The movement of cross sections of the monofilament nylon threads inserted into the axially loaded intervertebral disc was traced with magnetic resonance imaging (MRI). This technique allowed the observation of the sequential solid phase displacement of the loaded intervertebral disc. OBJECTIVES: To clarify sequential solid phase displacement of the axially loaded intervertebral disc to elucidate the cause of centripetal fluid shift within a disc. SUMMARY OF BACKGROUND DATA: We already have reported that there is a centripetal fluid shift within the axially loaded intervertebral disc during the early phase of loading. We assumed that there should be an elaborate intradiscal matrix displacement that generates a pressure gradient within the disc to cause a centripetal fluid shift. METHODS: Thirteen freshly obtained bovine caudal intervertebral discs were prepared. Three to five monofilament nylon threads were inserted into each disc in the anterior-posterior direction to trace the intradiscal solid phase displacement on the midcoronal MR images. Sequential displacement of the disc matrix was recorded during a 294 N axial loading. RESULTS: Relatively large centrifugal expansion at the inner layer of the anulus fibrosus compared with less centrifugal expansion of the outer anulus fibrosus was observed in accord with gradual creep of the disc thickness. CONCLUSIONS: The uneven displacement of the intradiscal solid phase observed in the present study expels the fluid phase from the inner anulus fibrosus, thus resulting in accumulation of fluid phase in the nucleus pulposus. The present study suggests the presence of a mechanism that retains water within the normal intervertebral disc, in spite of an external load, because it forms a water-abundant nucleus pulposus, which is surrounded by an anulus fibrosus with decreased water permeability caused by fluid loss. A more detailed analysis is required to clarify topographic volumetric changes within the disc.     

Histology of intervertebral disc protrusion: an experimental study using an aged rat model

STUDY DESIGN: In vitro experimental intervertebral disc ruptures of aged rats were examined histologically. OBJECTIVES: To clarify the mechanism of intervertebral disc herniations by microscopic investigation of ruptured discs. SUMMARY OF BACKGROUND DATA: Clinically, disc herniations have been classified into two types: extrusion and protrusion. However, the pathogenesis of protrusion type herniations has not yet been demonstrated by any studies. To clarify this issue, it is essential to establish an appropriate model producing disc herniations, and to examine the sequential changes in the structure of herniated discs. METHODS: Lumbar discs of 2-year-old rats were examined histologically and compared with human lumbar discs. To examine structural changes in discs subjected to repetitive motion stress, 400 repetitions of a sequence of flexion (30 degrees ) and axial rotation (6 degrees ) were applied in vitro to the lumbar discs of the animals. RESULTS: The microstructure of normal lumbar discs in aged rats was similar in many ways to the human lumbar discs in a 20- to 40-year-old adult. Of 10 discs subjected to repetitive stress, 4 were ruptured at the junction between the posterior anulus fibrosus and the sacral cartilage endplate. One had an extruded nucleus pulposus, and three had a protruded anulus fibrosus, which displayed disorganized structure containing widened and flaccid lamellae. CONCLUSIONS: The results from this study indicate that disc protrusion can be caused by disorganization of the ruptured annular lamellae, not by focal compression of the nucleus pulposus.     

Nucleus pulposus allograft retards intervertebral disc degeneration

Autogenous implantation of nucleus pulposus or nucleus pulposus cells that were activated by coculture retards intervertebral disc degeneration, but harvesting such grafts causes disc degeneration at the donor site. This study examined whether nucleus pulposus allografts similarly retard disc degeneration and whether such allografting induces immunologic rejection. Japanese White rabbits served as donors and recipients for allografts. Lumbar disc degeneration was induced by aspirating the nucleus pulposus. Two weeks later, intact nucleus pulposus or nucleus pulposus cells were injected and compared with a sham procedure and normal control. The recipients' discs were examined histologically and immunologically at intervals for 16 weeks. Discs receiving an intact nucleus pulposus showed the least degeneration, followed by discs receiving nucleus pulposus cells, both of which were better than no treatment. These findings correlated directly with the intensity of immunochemical staining for Type II collagen. Allogeneic grafts did not induce any appreciable host-versus-graft response. Injection of nucleus pulposus and nucleus pulposus cells retards intervertebral disc degeneration. However, injection of intact nucleus pulposus is more effective than injection of nucleus pulposus cells alone. The intercellular matrix plays an important, but poorly understood, role in preserving intervertebral discs.     

水通道蛋白3在人正常椎间盘中的表达

目的 观察水通道蛋白-3(aquaporin-3)在人正常椎间盘中的表达及分布.方法 收集10例青年人因腰椎骨折行椎间融合手术摘除的正常椎间盘样本,运用免疫组织化学、逆转录-聚合酶链反应(RT-PCR)、 Western blot检测AQP3在椎间盘中的表达及分布.结果 免疫组织化学显示AQP3表达于椎体软骨终板类软骨细胞,髓核和纤维环未见表达;RT-PCR显示AQP3 mRNA在椎体软骨终板及髓核组织有表达;Western blot检测显示椎间盘组织在29×103左右有一特异性条带.结论 AQP3在人正常椎间盘中的表达及分布提示其可能参与椎间盘内液体平衡,对维持椎间盘组织的正常生理功能、在椎间盘退变的发病机制中可能有重要作用.     

Histologic findings of disc, end plate and neural elements after coblation of nucleus pulposus: an experimental nucleoplasty study

BACKGROUND CONTEXT: Partial removal of the nucleus has been shown to decompress herniated discs, relieving pressure on nerve roots and, in some cases, offering relief from disc pain. The nucleoplasty technique builds on earlier surgical approaches that helped validate the strategy of intranuclear tissue removal. Nucleoplasty, a new minimally invasive procedure using patented coblation technology, combines coagulation and ablation for partial removal of the nucleus pulposus to decompress the disc. PURPOSE: To determine if histologic changes of the intervertebral discs and surrounding tissues occur after nucleoplasty. STUDY DESIGN: A light microscopic study of intervertebral disc and adjacent neural tissues after disc decompression by nucleoplasty in pig cadavers. METHODS: Light microscopy was used to examine disc and neural tissues in two pig cadaveric specimens (T12 to sacrum). Nucleoplasty was performed by 1) advancing a radiofrequency wand to a predetermined depth in the disc (ablation), and 2) withdrawing the wand to the starting point (coagulation). Discs and adjacent tissues were removed from treated and nontreated segments, and examined under light microscopy. RESULTS: Histologic examination revealed no evidence of direct mechanical or thermal damage to the surrounding tissues. There was clear evidence of coblation channels with clean coagulation borders of the nucleus pulposus. Normal histologic findings of the annulus and end plate, with normal neural elements of the spinal cord and nerve roots at the level of the procedure, were observed. CONCLUSIONS: The histologic findings of this study suggest that the nucleoplasty achieves volumetric removal of target disc tissue without overt thermal or structural damage to the adjacent tissues. Further studies in live animals will be needed to assess the effects of nucleoplasty on the annulus, end plate and neural tissues under physiologic conditions, including assessment of cell viability.     

Age-related changes in fibromodulin and lumican in human intervertebral discs.

STUDY DESIGN: An analysis of proteoglycans of the intervertebral disc using immunoblotting of tissue extracts. OBJECTIVES: To investigate the changes in structure and abundance of fibromodulin and lumican in human intervertebral discs during aging and degeneration. SUMMARY OF BACKGROUND DATA: Fibromodulin and lumican are keratan sulfate proteoglycan constituents of the disc's extracellular matrix, whose interaction with collagen fibrils may contribute to the mechanical properties of the tissue. Changes in their abundance and/or structure that occur with aging and degeneration therefore may have an impact on disc function. METHODS: Lumbar intervertebral discs were obtained from individuals of different ages, and extracts of anulus fibrosus and nucleus pulposus were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting using antibodies specific for fibromodulin and lumican. RESULTS: The major changes in abundance observed with age were a decrease in fibromodulin in the adult nucleus pulposus and an increase in lumican in anulus fibrosus during early juvenile development. In addition, fibromodulin in the anulus fibrosus exhibited a structural change with increasing age, characterized by a shift toward the predominance of its glycoprotein form lacking keratan sulfate. Fibromodulin was more abundant in the anulus fibrosus than in nucleus pulposus at all ages, whereas lumican was much more abundant in nucleus pulposus than in anulus fibrosus in the young juvenile; in the adult, however, lumican was present in comparable levels in both tissues. With increasing degrees of degeneration, fibromodulin exhibited an increase in abundance. CONCLUSIONS: Growth, aging, and degeneration of the intervertebral disc are associated with changes in the abundance and structure of fibromodulin and lumican, which presumably influence the functional properties of the tissue.     

Abnormal stress concentrations in lumbar intervertebral discs following damage to the vertebral bodies: a cause of disc failure?

Summary The purpose of this investigation was to test the hypothesis that damage to a lumbar vertebral body can lead to abnormal stress concentrations in the adjacent intervertebral discs. Twenty-three cadaveric lumbar motion segments, from persons who had died aged between 19 and 87 years, were subjected to substantial compressive loading while in the neutral, lordotic and flexed postures. During the loading period, a miniature pressure transducer was pulled through the disc along its mid-sagittal diameter and graphs of horizontal and vertical compressive stress against distance were obtained. Measurements were repeated after each motion segment had been compressed up to the point of mechanical failure: at this point the vertebral bodies suffered minor damage to the trabecular arcades, and sometimes to the end-plate, but the structure remained essentially intact and motion segment height was reduced by only 1%–2%. After damage, the stress in the nucleus and anterior annulus fell by about 30%, and high stress peaks appeared in the inner posterior annulus. These changes were more pronounced in lordotic posture and less pronounced in flexion. The youngest discs showed the smallest changes. It is concluded that minor compressive damage to the vertebral body can lead to high stress concentrations in the posterior annulus. Since the vertebral body is the weak link of the lumbar spine, this may be a frequent precipitating cause of isolated disc failure in living people.AcroMed Prize of the European Spine Society 1992     

Regional variations in the compressive properties of lumbar vertebral trabeculae. Effects of disc degeneration

The compressive mechanical properties of human lumbar vertebral trabeculae were examined on the basis of anatomic origin, bone density, and intervertebral disc properties. Trabecular bone compressive strength and stiffness increased with increasing bone density, the latter proportional to strength and stiffness to the one-half power. Regional variations within each segment were found, the most prevalent differences occurring in regions of bone overlying the disc nucleus in comparison with bone overlying the disc anulus. For normal discs, the ratio of strength of bone overlying the disc nucleus to bone overlying the disc anulus was 1.25, decreasing to 1.0 for moderately degenerated discs. These results suggest that an interdependency of trabecular bone properties and intervertebral disc properties may exist.     

Histology and pathology of the human intervertebral disc

The intervertebral disc is a highly organized matrix laid down by relatively few cells in a specific manner. The central gelatinous nucleus pulposus is contained within the more collagenous anulus fibrosus laterally and the cartilage end plates inferiorly and superiorly. The anulus consists of concentric rings or lamellae, with fibers in the outer lamellae continuing into the longitudinal ligaments and vertebral bodies. This arrangement allows the discs to facilitate movement and flexibility within what would be an otherwise rigid spine. At birth, the human disc has some vascular supply within both the cartilage end plates and the anulus fibrosus, but these vessels soon recede, leaving the disc with little direct blood supply in the healthy adult. With increasing age, water is lost from the matrix, and the proteoglycan content also changes and diminishes. The disc-particularly the nucleus-becomes less gelatinous and more fibrous, and cracks and fissures eventually form. More blood vessels begin to grow into the disc from the outer areas of the anulus. There is an increase in cell proliferation and formation of cell clusters as well as an increase in cell death. The cartilage end plate undergoes thinning, altered cell density, formation of fissures, and sclerosis of the subchondral bone. These changes are similar to those seen in degenerative disc disease, causing discussion as to whether aging and degeneration are separate processes or the same process occurring over a different timescale. Additional disorders involving the intervertebral disc can demonstrate other changes in morphology. Discs from patients with spinal deformities such as scoliosis have ectopic calcification in the cartilage end plate and sometimes in the disc itself. Cells in these discs and cells from patients with spondylolisthesis have been found to have very long cell processes. Cells in herniated discs appear to have a higher degree of cellular senescence than cells in nonherniated discs and produce a greater abundance of matrix metalloproteinases. The role that abnormalities play in the etiopathogenesis of different disorders is not always clear. Disorders may be caused by a genetic predisposition or a tissue response to an insult or altered mechanical environment. Whatever the initial cause, a change in the morphology of the tissue is likely to alter the physiologic and mechanical functioning of the tissue.     

Cyclic mechanical stretch stress increases the growth rate and collagen synthesis of nucleus pulposus cells in vitro

STUDY DESIGN: A rabbit model designed to investigate the effects of applied cyclic tensile stress on the cell division rate and the collagen synthesis in the rabbit nucleus pulposus cells in vitro. OBJECTIVE: To evaluate the effects of mechanical stress on nucleus pulposus cells, thus adding to the understanding of the adaptation of the intervertebral disc to mechanical stress. SUMMARY OF BACKGROUND DATA: Intervertebral disc cells in vivo are exposed to a multitude of physical forces during physical motion. Although it is known that in intervertebral disc disease, a common pathway of disc degeneration is mechanical stress on the nucleus pulposus or the anulus fibrosus or both, the underlying mechanism has been less well defined. METHODS: Nucleus pulposus cells were isolated from 4-week-old Japanese white rabbits. These cells were subjected to the mechanical cyclic stretch stress using a computerized, pressure-operated instrument that physically deformed the cells. The DNA synthesis rate, collagen synthesis rate, and cell cycle progression were measured. RESULTS: Cyclic tensile stretch increased the DNA synthesis rate in nucleus pulposus cells and in the population of cells in the S phase of the cell cycle during 1 to 2 days of subjugation to stress. Cyclic tensile stretch also increased collagenous protein synthesis in nucleus pulposus cells during 1 to 4 days of stress. CONCLUSIONS: Mechanical stress on nucleus pulposus cells promotes the proliferation of cells and alters the properties of intervertebral disc cells. This study may reflect the adaptation of the intervertebral disc to increased motion and stress.     

Contribution of disc degeneration to osteophyte formation in the cervical spine: a biomechanical investigation.

Cervical spine disorders such as spondylotic radiculopathy and myelopathy are often related to osteophyte formation. Bone remodeling experimental-analytical studies have correlated biomechanical responses such as stress and strain energy density to the formation of bony outgrowth. Using these responses of the spinal components, the present study was conducted to investigate the basis for the occurrence of disc-related pathological conditions. An anatomically accurate and validated intact finite element model of the C4-C5-C6 cervical spine was used to simulate progressive disc degeneration at the C5-C6 level. Slight degeneration included an alteration of material properties of the nucleus pulposus representing the dehydration process. Moderate degeneration included an alteration of fiber content and material properties of the anulus fibrosus representing the disintegrated nature of the anulus in addition to dehydrated nucleus. Severe degeneration included decrease in the intervertebral disc height with dehydrated nucleus and disintegrated anulus. The intact and three degenerated models were exercised under compression, and the overall force-displacement response, local segmental stiffness, anulus fiber strain, disc bulge, anulus stress, load shared by the disc and facet joints, pressure in the disc, facet and uncovertebral joints, and strain energy density and stress in the vertebral cortex were determined. The overall stiffness (C4-C6) increased with the severity of degeneration. The segmental stiffness at the degenerated level (C5-C6) increased with the severity of degeneration. Intervertebral disc bulge and anulus stress and strain decreased at the degenerated level. The strain energy density and stress in vertebral cortex increased adjacent to the degenerated disc. Specifically, the anterior region of the cortex responded with a higher increase in these responses. The increased strain energy density and stress in the vertebral cortex over time may induce the remodeling process according to Wolff's law, leading to the formation of osteophytes.     

Viscoelastic properties of intervertebral disc cells. Identification of two biomechanically distinct cell populations

STUDY DESIGN: A combined experimental and theoretical biomechanical study to quantify the mechanical properties of living cells of the porcine intervertebral disc. OBJECTIVES: To quantify zonal variations in the mechanical properties and morphology of cells isolated from the intervertebral disc. SUMMARY OF BACKGROUND DATA: Cellular response to mechanical stimuli is influenced by the mechanical properties of cells and of the extracellular matrix. Significant zonal variations in intervertebral disc matrix properties have been reported. No information is currently available on the corresponding regional variations in the mechanical properties of intervertebral disc cells, despite evidence of significant differences in cellular phenotype and biologic response to loading. METHODS: The micropipette aspiration test was used in combination with a three-parameter viscoelastic solid model to measure the mechanical properties of cells isolated from the anulus fibrosus, transition zone, and nucleus pulposus. RESULTS: Intervertebral disc cells exhibited viscoelastic solid behaviors. Highly significant differences were observed in the morphology, cytoskeletal arrangement, and biomechanical properties of the nucleus pulposus cells as compared with anulus fibrosus or transition zone cells. Cells of the nucleus pulposus were approximately three times stiffer and significantly more viscous than cells of the anulus fibrosus or transition zone. CONCLUSIONS: The findings of this study provide new evidence for the existence of two biomechanically distinct cell populations in the intervertebral disc. These differences in mechanical behavior may be related to observed differences in the cytoskeletal architecture between these cells, and may further play an important role in the development, maintenance, and degeneration of the intervertebral disc.     

Induced narrowing and back adaptation of lumbar intervertebral discs in biomechanically stressed rats.

The purpose of this work was to develop an animal model of chronic low-back dysfunction induced by biomechanical stress. The intervertebral disc size was measured by radiography and histology in control rats, in rats intermittently forced to ambulate on a flat surface or in cylinders. Forced ambulation in cylinders caused rats to assume a relative extensive posture in the low back. Lumbar discs in biomechanically stressed rats initially narrowed, and after months of continued intermittent stress, regained near-normal size in vivo, shown by radiography. By using histologic methods, discs from rats that ambulated in any posture were shown to be larger than discs from rats not forced to ambulate. The results were consistent with the hypothesis that in vivo discs in biomechanically stressed rats initially narrow due to compressive loading and then widen back toward normal size because of an altered matrix which has increased imbibing properties. According to this hypothesis altered matrix synthesis in the nucleus pulposus was an adaptation induced by the cyclic compressive loading associated with ambulation.     

Phenotypic characteristics of rabbit intervertebral disc cells. Comparison with cartilage cells from the same animals.

STUDY DESIGN: Intervertebral disc cells were extracted from the surrounding matrix, and their metabolic activities and phenotypes were studied. OBJECTIVES: To compare the metabolic activities and phenotypes of cell populations extracted from the intervertebral discs of young rabbits with those of articular and growth plate chondrocytes from the same animals. SUMMARY OF BACKGROUND DATA: The phenotype of intervertebral disc cells has been poorly studied and still is debated. METHODS: The intervertebral discs as well as articular and vertebral growth plate cartilage of rabbits were digested enzymatically. The morphology of freshly isolated cells was examined. Their contents of collagen II and X mRNAs were determined by Northern blot analysis, and their sulfation activity by 35S-sulfate incorporation as chondrocytic markers. Cells were cultured at high density or low density and grown in primary culture. The stability of their phenotype was monitored by evaluating the collagen I and II mRNA ratio. The proteoglycans newly synthesized by the cells also were quantified, and their elution profile analyzed on Sepharose 2B columns. RESULTS: The anulus fibrosus cells were morphologically undistinguishable from articular chondrocytes. The nucleus pulposus contained mainly large vacuolated cells and a few smaller cells. All freshly extracted cells expressed different levels of collagen II mRNA. Anulus fibrosus and nucleus pulposus cells contained, respectively, 22% and 8% of collagen II mRNA compared with that found in articular or growth plate chondrocytes from the same animal. Only growth plate chondrocytes expressed collagen X. When anulus fibrosus cells were incubated for 48 hours at high density, they had collagen II mRNA contents similar to those of articular and growth plate chondrocytes, but synthesized five to six times fewer sulfated proteoglycans. When seeded at low density, anulus fibrosus cells divided more slowly than articular chondrocytes and incorporated four times fewer 35S-sulfate into proteoglycans. Their collagen II mRNA content was 2.75-fold lower than that of chondrocytes, and the procollagen alpha 1II/alpha 1I mRNA ratio was 3.1 for anulus fibrosus cells and 7 for chondrocytes. No collagen X mRNA was detected. When incubated for 48 hours at high density, the nucleus pulposus giant cells had four times less collagen II mRNA content than cartilage cells but synthesized the same amounts of sulfated proteoglycans. They did not divide during 21 days in culture and still contained collagen II mRNA but no collagen X mRNA. CONCLUSIONS: Findings showed that intervertebral disc cells all express cartilage-specific matrix proteins with quantitative differences, depending on their anatomic situation. It is suggested that anulus fibrosus cells are chondrocytic cells at a different stage of differentiation than articular and growth plate chondrocytes. The phenotype of nucleus pulposus cells still is unclear. They could be chondrocytic or notochordal. A definitive answer to this important question requires differentiating markers of notochordal cells.