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PubMed1983

Target tissue models: the proteoglycans and degeneration of the human intervertebral disc.

Seventy-five intervertebral discs from 15 spines were analyzed after morphologic grading. With degeneration, the water and proteoglycan (PG) contents decreased and the collagen content increased. The PG was isolated from 20 discs by single-stage dissociative density gradient ultracentrifugation. Neither the chondroitin 6-sulphate nor the protein contents of the PG changed with degeneration although the keratan sulphate fell slightly.

Collagen↗
PubMed1991

Lumbar intervertebral foramens. An in vitro study of their shape in relation to intervertebral disc pathology.

The lumbar intervertebral foramens of 20 isolated cadaveric spines were investigated by the use of a molding technique to assess their dimensions accurately and how intervertebral disc pathology altered their configuration. Oval foramens predominated over auricularly shaped foramens when the disc was normal, but when the disc was abnormal, the converse was true. Foraminal size varied from 40 to 160 mm2 but showed great variation even at individual levels; these were not accurately reflected by simple radiologic measurements.

Adolescent↗
PubMed2001

The relationship between apoptosis of endplate chondrocytes and aging and degeneration of the intervertebral disc.

STUDY DESIGN: Apoptosis in cervical intervertebral disc cells and cartilaginous endplate cells was examined by the nick end labeling (TUNEL) technique during the process of natural aging and in a mouse experimental spondylosis model. OBJECTIVES: To determine the role of apoptosis in aging and degeneration of intervertebral discs by monitoring chronologic changes in the quantity and localization of apoptotic cells. SUMMARY OF BACKGROUND DATA: Apoptosis occurs within human intervertebral discs, but little is known about the pathologic significance of this process. On the other hand, the cartilaginous endplate is known to decrease in thickness and to disappear with aging and degeneration. The cause of this age-related change remains unclear. METHODS: A mouse spondylosis model was prepared via surgical resection of the posterior spinal element in 12 mice to examine the experimentally induced spondylosis process. Eighteen naturally aged mice were also used to examine the influence of aging. Paraffin-embedded midsagittal sections of the cervical spine were obtained 2, 3, 6, and 12 months after surgery in the spondylosis model and in the age-matched naturally aged mice, as well as in 4-week-old and 18-month-old naturally aged mice. Sections were stained with hematoxylin and eosin, safranin-O, and the TUNEL procedure. The number of apoptotic cells and vital cells were counted in the cartilaginous endplate of the intervertebral disc excluding the growth cartilage, and the degree of disappearance of the cartilaginous endplate was evaluated. RESULTS: Apoptosis, particularly noticeable in the cartilaginous endplate, increased with age and resulted in a marked decrease in cell density. Subsequently, the structure of the cartilaginous endplate began to disappear. Apoptosis was more evident and the structure of the cartilaginous endplate began to disappear more rapidly in the surgically treated group than in the naturally aged group. CONCLUSIONS: TUNEL-positive cells in the cartilaginous endplate increased with age, with destruction of the cartilaginous endplate after apoptosis (TUNEL-positive cell death). The application of the spondylosis model increased the incidence of apoptosis preceding the development of spondylosis. This suggests that apoptosis plays a role in the age-related changes seen in the cartilaginous endplate of the intervertebral disc and in the experimentally induced spondylosis process.

Aging↗
PubMed1998

Congenital maldevelopment of intervertebral disc simulating a neurofibroma.

Intervertebral disc herniation is a common cause of radiculopathy and myelopathy in adulthood. It is an uncommon tumor mimic. We report on an extradural disc associated with an osseous defect ostensibly caused by pressure erosion and appearing as a neural tumor. It showed homogeneous enhancement on a contrast-enhanced MR examination, leading to an erroneous diagnosis of nerve sheath tumor. An attempt has been made to explain the likely mechanism of formation accounting for the imaging appearances, along with a review of the literature.

Diagnosis, Differential↗
PubMed2000

Histological study of lumbar intervertebral disc herniation in adolescents.

Herniated intervertebral discs are rare in children and adolescents constituting approximately 1-5% of all patients undergoing surgery for lumbar and lumbosacral intervertebral disc herniation. Preceding traumata and congenital anomalities have been reported as important factors for the pathogenesis of intervertebral disc prolapses in young patients. The present histological study is based upon 15 patients with lumbar disc herniation within an age range from 14 to 19 years. Only in one case, was adequate trauma reported. All patients exhibited degenerative changes of the disc, similiar to those observed in adults. These changes were marked in 11 patients (73%). Thus, as known from adults, also in isolated traumatic disc herniation of adolescence, pre-existing degeneration of the disc has to be considered. If such changes are present, trauma has the significance of only transitory deterioration of the previous disc degeneration.

Adolescent↗
PubMed2002

Fas ligand exists on intervertebral disc cells: a potential molecular mechanism for immune privilege of the disc.

STUDY DESIGN: Rat and human intervertebral disc specimens were examined immunohistochemically. Reverse transcription polymerase chain reaction (RT-PCR) analysis was also performed on rat disc tissue to demonstrate the existence of Fas ligand. OBJECTIVE: To clarify the existence of Fas ligand on intact intervertebral disc cells. SUMMARY OF BACKGROUND DATA: The nucleus pulposus has been reported to be an immune-privileged site. The immune-privileged characteristic in other tissues such as the retina and testis has been attributed to the local expression of Fas ligand, which acts by inducing apoptosis of invading Fas-positive T-cells. The existence of Fas ligand in normal disc cells has not yet been addressed. METHODS: Skeletally mature SD male rats were killed, and the coccygeal discs were harvested. Human disc specimens were obtained from idiopathic scoliosis patients during surgical procedures. Immunohistochemical staining for Fas ligand was performed for cross-sections of the discs by standard procedures. Reverse transcription polymerase chain reaction analysis was also carried out to demonstrate Fas ligand mRNA expression on rat intervertebral discs. Testes of the rats were used for positive controls, and muscles were used for negative controls. The sections were observed by light microscopy. RESULTS: The nucleus pulposus cells exhibited intense positive immune staining for Fas ligand. The outer anulus fibrosus cells and notochordal cells exhibited little immunopositivity. The positive controls exhibited positive immune staining, and the negative control showed no immunopositivity. The result of RT-PCR confirmed the existence of Fas ligand in disc cells. The human nucleus pulposus cells showed a similar predilection to rat disc cells. CONCLUSIONS: We demonstrated the existence of Fas ligand on disc cells, which should play a key role in the potential molecular mechanism to maintain immune privilege of the disc. Immune privilege and Fas ligand expression of the intervertebral disc may provide a new insight for basic science research as well as clinical treatments for disc degenerative diseases, including disc herniation with radicular pain.

Animals↗
PubMed2006

Correlation of swelling pressure and intrafibrillar water in young and aged human intervertebral discs.

Fluid balance in the intervertebral disc under applied load is determined primarily by its swelling pressure, that is, the external pressure at which it neither loses nor gains water. This depends on the composition of the tissue, in particular on its proteoglycan concentration. Proteoglycans develop a high osmotic pressure due to their fixed negatively charged groups. Because of their size, proteoglycans are excluded from the collagen's intrafibrillar volume; hence their osmotic activity is determined only by the extrafibrillar water. Here, we show that in order to evaluate correctly the swelling pressure in the annuli fibrosi of human intervertebral disc, it is essential to evaluate its proportion of intrafibrillar water. We used low-angle X-ray scattering and osmotic stress techniques to determine the lateral packing of the collagen molecules in the fibrils of the annuli fibrosi (ages: 25-77). It was found that the lateral packing and, hence, the intrafibrillar water content depends on age, external osmotic pressure, and location in the tissue. Subtracting intrafibrillar water from total hydration yields the amount of extrafibrillar water, from which the true fixed charge density of the tissue could be estimated. From a force balance, it would appear that collagen tension plays only a minor role in the equilibrium of the human intervertebral disc under load, in contrast to articular cartilage, where collagen tension is important for load bearing.

Adult↗
PubMed1994

[Laser ablation of intervertebral disc: animal experiment].

The lumbar intervertebral discs (L3-6) were ablated through a transperitoneal approach in 12 adult dogs by using Nd: YAG laser (1.06 microns) with a 600 microns quartz fiber. The status of limbs motion and sphincter (bladder, bowel) was observed for evaluating the safety of laser irradiation. After irradiation, the animals were sacrificed at prescribed intervals of up to 40 weeks (2, 4, 8, 12 and 40 weeks after operation). The lumbar intervertebral discs were harvested and subjected to light microscopic observation. No dog had suffered from neurogenic dysfunction of limb motion and sphincter. Histological findings immediately after the irradiation showed the disc was vaporized and a cavity was made. After 2 and 4 weeks, fibrous tissues began to proliferate, but cartilaginous tissues replaced the fibrous tissues 12 weeks after the laser irradiation. No new bone formation was found within 40 weeks after operation. On the basis of this study and our previous cadaveric study, percutaneous laser disc decompression (PLDD) was applied in clinical practice in march of 1993. 10 patients underwent PLDD utilizing the same laser equipment. The average follow-up was 3 months. According to the Macnab's criteria, there was an excellent response in 7 patients and a good response in 3.

Animals↗
PubMed2000

Does a herniated nucleus pulposus contribute significantly to a decrease in height of the intervertebral disc? Quantitative volumetric MRI.

A lumbar intervertebral disc with a herniated nucleus pulposus (HNP) often exhibits a decrease in the height of the intervertebral space. Our purpose was to ascertain whether the loss of volume of an HNP is sufficient to cause a perceptible decrease in the height of the intervertebral space. MRI of 44 patients with 51 HNPs were reviewed. The volumes of the herniated material and of the intervertebral discs were calculated for every level from L1-2 to L5-S1. The average volume of the HNP was 503 +/- 301 mm3. The average volumes of all 220 intervertebral discs and of the 127 normal-appearing discs were 14,442 +/- 4200 mm3 and 17,476 +/- 2885 mm3 respectively. The average volume of the HNP represented 3.5% of the parent disc. An average HNP caused a decrease in intervertebral space height of 0.35 mm (0.56 pixels). Therefore, the loss of the volume of the HNP does not cause a significant decrease in the intervertebral space height. The average calculated decrease in the disc height is less than that reported in normal diurnal variation.

Adult↗
PubMed2006

Advanced glycation end-products downregulating intervertebral disc cell production of proteoglycans in vitro.

OBJECT: The authors sought to clarify the role, if any, of advanced glycation end-products (AGEs) in disc degeneration. METHODS: Intervertebral discs were analyzed for the presence of AGEs and of their receptor (RAGE) by immunohistochemical analysis. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to detect any RAGE gene expression, and real-time PCR was used to quantify messenger RNA (mRNA) levels of aggrecan and collagen types I and II in nucleus pulposus cells treated with AGEs. Aggrecan protein concentration was determined by enzyme-linked immunosorbent assay. Immunohistochemical analysis revealed that AGEs and RAGE were localized in the nucleus pulposus of the intervertebral disc. Advanced glycation end-products were found to significantly suppress the expression of aggrecan at both mRNA and protein levels in a dose- and time-dependent manner. The levels of collagen types I and II remained unchanged after treatments with AGEs. CONCLUSIONS: These results suggest that the accumulation of AGEs and their interaction with their receptor in the nucleus pulposus might result in the downregulation of aggrecan production responsible for disc degeneration.

Aged↗
PubMed1991

Histochemical and ultrastructural observations on brown degeneration of human intervertebral disc.

Thirty-eight fresh human intervertebral discs collected during anterior interbody fusion surgery were histochemically and ultrastructurally analyzed for pigments. Macroscopically, five stages of degeneration were classified according to the color, fibrosis, and fragility of the nucleus pulposus of the discs. In order to demonstrate lipofuscin granules, specimens were subjected to special staining procedures, including carbol fuchsin lipofuscin stain, the Schmorl's reaction, and autofluorescence. Lipofuscin granules were distributed from the inner layer of the annulus fibrosus to the nucleus pulposus. Such granules were numerous in cases of slight or severe degeneration, whereas fewer granules were found in cases of moderate degeneration. However, the stage of macroscopic degeneration of the intervertebral disc did not necessarily correlate with the incidence of lipofuscin granules. By ultrastructural observation, the morphological features of the components of the intervertebral disc and the ultrastructure of the lipofuscin granule were clarified. The ultrastructure of the "brown degeneration" disc exhibited markedly increased amorphous electron-dense bodies located among collagen fibrils in the matrix.

Adult↗
PubMed2002

Spontaneous production of monocyte chemoattractant protein-1 and interleukin-8 by the human lumbar intervertebral disc.

STUDY DESIGN: Scoliotic and herniated human intervertebral disc tissue obtained intraoperatively was cultured, and the medium was analyzed for the production of monocyte chemoattractant protein-1 and interleukin-8. OBJECTIVES: This study was conducted to determine whether the human intervertebral disc is capable of spontaneous production of the chemokines monocyte chemoattractant protein-1 and interleukin-8. SUMMARY OF BACKGROUND DATA: Lumbar disc herniations undergo spontaneous regression with time. This is believed to occur via macrophage-mediated phagocytosis of herniated disc material. Monocyte chemoattractant protein-1, a chemotactic agent for macrophages, has recently been identified in rat intervertebral disc tissue. METHODS: Disc material obtained from patients undergoing surgery for scoliosis and sciatica was cultured using a serumless technique, and the medium was subsequently analyzed for levels of monocyte chemoattractant protein-1 and interleukin-8. RESULTS: Monocyte chemoattractant protein-1 and IL-8 were detected in both control and herniated disc specimens. Noncontained herniations produced higher levels of chemokines than those with an intact anulus. CONCLUSIONS: Human intervertebral disc tissue is capable of spontaneously producing the proinflammatory chemokines monocyte chemoattractant protein-1 and interleukin-8. These are chemotactic for macrophages and capillaries and may explain the ingrowth of granulation tissue seen in spontaneous disc herniation resorption.

Adolescent↗
PubMed2000

Effect of anular repair on the healing strength of the intervertebral disc: a sheep model.

STUDY DESIGN: The intervertebral disc, in a sheep model, was used to assess the effect of directly repairing three different anular incisions on the subsequent healing strength of the intervertebral disc. OBJECTIVES: To assess whether directly repairing an anular defect, made at the time of lumbar discectomy, could influence the healing rate and strength of the anulus fibrosus. METHODS: Twenty-four sheep underwent a retroperitoneal approach to five lumbar disc levels. An anular incision, followed by partial discectomy was done at each exposed level. Anular incisions used in this study consisted of 1) a straight transverse slit, 2) a cruciate incision, and 3) a window or box excision. Healing strength was measured at three time intervals: 2 weeks, 4 weeks, and 6 weeks. Each anular incision type was performed on 30 lumbar discs, 10 discs in each time interval. Five discs in each time interval underwent direct repair, and five discs were left unrepaired to heal as controls. The sheep were killed at 2, 4, and 6 weeks after surgery. The lumbar spines were removed en bloc, and the intervertebral discs were subjected to pressure-volume testing to assess the anular strength of repaired versus unrepaired disc injuries at each time interval. RESULTS: Statistical analysis was performed to evaluate the effects of healing time, incision technique, and repair on the pressure-volume characteristics of the involved discs. Pressure-volume testing showed trends of stronger healing for repaired discs, but at no time interval was any significant difference found between repaired and nonrepaired anular strength. Of the nonrepaired discs, the box incision was only 40 to 50% as strong as the slit or cruciate incised discs during early healing. CONCLUSION: Direct repair of anular incisions in the lumbar spine does not significantly alter the healing strength of the intervertebral disc after lumbar discectomy.

Animals↗
PubMed2003

Disc chondrocyte transplantation in a canine model: a treatment for degenerated or damaged intervertebral disc.

STUDY DESIGN: Disc degeneration and osteoarthritis are diseases of the matrix. Chondrocytes that have been removed from damaged cartilaginous tissues maintain a capacity to proliferate, produce, and secrete matrix components, and respond to physical stimuli such as dynamic loading. A dog model was used to investigate the hypothesis that autologous disc chondrocytes can be used to repair damaged intervertebral disc. OBJECTIVES: Given the capacity for the cells in vitro to produce matrix molecules that would be appropriate for disc chondrocytes, the focus of the experiment was to investigate whether the cells would continue to sustain metabolic function after transplantation. SUMMARY OF THE BACKGROUND DATA: No evidence for long-term integration exists for cell transplantation in species other than rats and rabbits. Furthermore, no controlled studies of 1-year duration have been published. MATERIALS AND METHODS: Disc chondrocytes were harvested and expanded in culture under controlled and defined conditions, returned to the same animals from which they had been sampled (autologous transplantation) via percutaneous delivery. The animals were analyzed at specific times after transplantation by several methods to examine whether disc chondrocytes integrated with the surrounding tissue, produced the appropriate intervertebral disc extracellular matrix, and might provide a formative solution to disc repair. RESULTS: In the context of degenerative changes in an injury model: (1) autologous disc chondrocytes were expanded in culture and returned to the disc by a minimally invasive procedure after 12 weeks; (2) disc chondrocytes remained viable after transplantation as shown by Bromodeoxyuridine incorporation and maintained a capacity for proliferation after transplantation as depicted by histology; (3) transplanted disc chondrocytes produced an extracellular matrix that displayed composition similar to normal intervertebral disc tissue. Positive evidence of proteoglycan content was supported by accepted histochemical staining techniques such as Safranin O-Fast Green; (4) both type II and type I collagens were demonstrated in the regenerated intervertebral disc matrix by immunohistochemistry after chondrocyte transplantation; and (5) when the disc heights were analyzed for variance according to treatment, a statistically significantcorrelation between transplanting cells and retention of disc height was achieved. CONCLUSIONS: Autologous chondrocyte transplantation is technically feasible and biologically relevant to repairing disc damage and retarding disc degeneration.

Animals↗
PubMed1994

Intervertebral disc amyloidosis: histochemical, immunohistochemical and ultrastructural observations.

Intervertebral discs selected at random from autopsies and surgical operations on herniated discs were examined for evidence of amyloid deposition. Amyloid deposits were detected in 53 (81.5%) of 65 autopsy cases. Discs from subjects over 50 years of age revealed a significantly high incidence of amyloid deposition. Among herniated discs, amyloid deposits were documented in 49 (75.4%) of 65 surgical specimens. The incidence of amyloid deposition in intervertebral discs increased with advancing age. Intervertebral disc amyloidosis consisted of amyloid deposits of three morphological types: linear amyloid deposits around the degenerating chondrocytes (perichondrocyte type), and nodular or band-like deposits in the annulus fibrosus (annulus fibrosus type) and nucleus pulposus (nucleus pulposus type). We suspect that the precursor protein of perichondrocyte type amyloid is derived from chondrocytes, and those of annulus fibrosus type and nucleus pulposus types are derived from the blood. The amyloid deposits were resistant to treatment with potassium permanganate. Immunohistochemically, the amyloid deposits reacted with antibody against amyloid P-component, but not with antibodies against AA, A kappa, A lambda, transthyretin or beta 2-microglobulin. Ultrastructurally, the amyloid deposits were composed of 10 nm-wide nonbranching fibrils. The amyloid in herniated discs had the same biochemical and immunohistochemical properties as those found in autopsy cases. The immunohistochemical characteristics of the amyloid deposits suggest that it derives from an, as yet, unknown precursor protein.

Adolescent↗
PubMed1999

Effects of basic fibroblast growth factor on spontaneous resorption of herniated intervertebral discs. An experimental study in the rabbit.

STUDY DESIGN: Histologic examination was performed on the autologous intervertebral disc material that was removed from the intervertebral space at L1-L2 and then relocated to the L4 posterior epidural space after the addition of basic fibroblast growth factor (bFGF) in a rabbit. OBJECTIVES: To evaluate whether basic fibroblast growth factor influences the resorption process of the herniated intervertebral disc through the promotion of angiogenesis and chemotaxis. SUMMARY OF BACKGROUND DATA: It has been reported that newly formed vessels, inflammatory cells, and their products may play an important role in the spontaneous resorption process of herniated intervertebral discs. In a rabbit model that mimics the sequestration type of intervertebral disc herniation, it has been reported that the autologous intervertebral disc material that relocated into the epidural space was penetrated by newly formed vessels originating from the epidural fat tissue. Therefore, it is possible that promotion of angiogenesis may influence the resorption of herniated intervertebral discs. Basic fibroblast growth factor is well known as an angiogenesis stimulation factor in vivo. METHODS: Thirty-six adult rabbits were divided into three groups. The L1-L2 intervertebral disc was partially incised through a retroperitoneal approach in each rabbit. The harvested disc material, which contained nucleus pulposus and anulus fibrosus, was immersed in one of three kinds of solution before relocation into the posterior epidural space at L4. In the control group, the harvested intervertebral disc was immersed in physiologic saline for 2 hours before relocation. In the group receiving 5 micrograms bFGF, the disc was immersed in 5 micrograms/mL bFGF for 2 hours before the relocation. In the group receiving 20 micrograms bFGF, the disc was immersed in 20 micrograms/mL bFGF for 2 hours before the relocation. Rabbits of each group were killed for histologic examination 1, 2, 4, and 8 weeks after surgery. RESULTS: In the bFGF-treated groups, newly formed vessels were observed to be in more numerous than those in the control group, 1 and 2 weeks after surgery. The number of inflammatory cells, including macrophages, lymphocytes, and fibroblasts, also increased in the bFGF-treated groups. The period from the surgery to the degradation of the intervertebral disc in the bFGF-treated groups was shorter than that in the control group, although the resorption process of the relocated discs was also observed in the control group. The size of relocated intervertebral discs in the bFGF-treated groups decreased at a higher rate than in the control group as time progressed. The rate of decrease in the size of discs in the group treated with 20 micrograms bFGF was more than that in the group treated with 5 micrograms. CONCLUSIONS: Epidural injection of bFGF facilitated the resorption of the intervertebral disc relocated to the epidural space.

Animals↗
PubMed1991

Juvenile intervertebral disc calcification: recognition, management, and pathogenesis.

Juvenile intervertebral disc calcification is an uncommon disorder of childhood, characterized by calcification of the nucleus pulposus of one or more intervertebral discs. Calcification may remain dormant or subsequently become symptomatic. The symptoms include fever, malaise, and neck pain and are associated with an elevated erythrocyte sedimentation rate and, occasionally, leukocytosis. Although disc protrusion occurs in 38% of patients, neurological signs are distinctly uncommon. We report the case of a patient with a herniated T2-T3 calcified intervertebral disc and compressive myelopathy. Juvenile intervertebral disc calcification is generally a self-limiting disease that seldom requires an operation. The symptoms are transient, and resorption of the disc calcification is the rule once symptoms occur. Neither the cause of the disc calcification nor the trigger for the onset of symptoms is known. An inflammatory response within the disc appears to give rise to clinical symptoms and is associated with eventual resorption of the disc calcification.

Calcinosis↗
PubMed2006

Relevance of in vitro and in vivo models for intervertebral disc degeneration.

Models available for the study of intervertebral disc degeneration are designed to answer many important questions. In vitro biologic models employ a variety of cell, tissue, or organ culture techniques with culture conditions that partially mimic the cellular environment of the degenerated human intervertebral disc. In vitro biomechanical models include intervertebral disc or motion-segment loading experiments as well as finite element modeling techniques. The literature describes numerous in vivo animal models for use in the study of intervertebral disc degeneration, each of which has its own advantages and disadvantages. Human-subject studies have included the use of magnetic resonance imaging and other techniques to assess diffusion into the intervertebral disc, to measure intradiscal pressure, to conduct kinematic or stiffness studies of lumbar motion segments, and to evaluate muscular forces on the spine. Although all of these studies are helpful in answering specific questions, their relevance in assessing disc degeneration in patients with symptoms of discogenic pain must be carefully considered.

Age Factors↗