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Basing on preliminary endoscopic examinations to present the intervertebral disc interspace using a common arthroscope in operations performed for lumbar disc prolapses, a special endoscopic instrumentarium (nucleoscope Hausmann-Forst) was developed allowing detailed and riskless views into the intervertebral disc interspace.
STUDY DESIGN: This study is an in vitro investigation of the effects of substance P on intervertebral disc cell metabolism. OBJECTIVES: To determine whether the neuropeptide, substance P, affects cells isolated from the intervertebral disc. SUMMARY OF THE BACKGROUND DATA: Nerve fibers containing substance P are present in the anulus fibrosus and may be released from the nerve terminals as in other tissues. Substance P is mitogenic for a variety of immune and connective tissue cells, and a fragment of the peptide affects the metabolism of articular chondrocytes. METHODS: Cells were isolated enzymically from the anulus fibrosus of intervertebral disc of 8-week-old rabbits. The effects of substance P and the C-terminal pentapeptide fragment SP7-11 on cell proliferation and proteoglycan deposition were determine by crystal violet and Alcian blue staining, respectively. RESULTS: Substance P ((10)-11-(10)-7 mol/l) had a small stimulatory effect on disc cell proliferation. Proteoglycan deposition in the cell layer increased concomitantly. A greater proliferative effect was observed with substance P fragment 7-11 or with the addition of the neutral endopeptidase inhibitor, phosphoramidon. CONCLUSIONS: Substance P has small mitogenic effects on rabbit intervertebral disc cells in vitro. Further investigation is required to establish whether this might have biologic relevance in relation to the maintenance or repair of the intervertebral disc.
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.
STUDY DESIGN: Biochemical study of human intervertebral discs collected at surgery from patients with low back pain associated with disc degeneration or scoliosis. Matrix metalloproteinases were studied by quantitative zymography. OBJECTIVE: To determine whether changes in the expression of matrix metalloproteinases will bring about tissue remodelling that contributes to the progressive nature and pathology of these diseases of the intervertebral disc. SUMMARY OF BACKGROUND DATA: The diseases of the intervertebral disc, degenerative disc disease and scoliosis, are both characterized by changes in the extracellular matrix components that will affect the mechanical function of the tissue. Matrix metalloproteinases are known to have the capability of degrading all the known extracellular matrix components of the disc. METHODS: Matrix metalloproteinases 2 and 9 were detected by gelatin-gel zymography and quantified by laser scanning densitometry. Both pro and active forms of the enzymes were measured. Thirty-four discs from patients with low back pain and 29 from patients with scoliosis were investigated. RESULTS: A correlation was found between the increasing levels of matrix metalloproteinases 2 and 9 and the grade of degenerative disc disease. In addition, the levels of these enzymes show a differential expression across the scoliotic disc with the highest levels in samples taken from the convexity of the curve. CONCLUSIONS: The difference between the concave and convex side of the scoliotic curve indicates that mechanical loads might influence the expression of these enzymes. The increased expression of these enzymes in both degenerative disc disease and scoliosis strongly suggests that they may affect the progressive nature of these diseases.
BACKGROUND: The effect of the menopause and HRT on the intervertebral discs has not been investigated. METHODS: One hundred women were recruited, comprising of 44 post-menopausal women on HRT, 33 untreated post-menopausal women and 23 pre-menopausal women. The height of the intervertebral discs between the 12th thoracic vertebra and the 3rd lumbar vertebra was measured by utilizing the bone densitometer height cursors. RESULTS: The untreated menopausal group of women had the lowest total disc height (D1-D3: 1.95 0.31 cm). This was significantly lower than the pre-menopausal group D1-D3: 2.16 0.24 cm) and the hormone-treated group (2.2 0.26 cm) (P > 0.02). The 2nd intervertebral disc consistently maintained a significant difference between the untreated menopausal group (D2: 0.63 0.13) and the other two groups (pre-menopausal group (D2: 0.72 0.09 cm) and treated menopausal group (D2: 0.73 0.12 cm) (P > 0.02). CONCLUSIONS: Estrogen-replete women appear to maintain higher intervertebral discs compared to untreated post-menopausal women. The estrogenic milieu may be relevant because of the significant impact it has on the hydrophilic glycosaminoglycans, the water content, collagen and elastin of the intervertebral discs. The maintenance of adequate disc height may allow the intervertebral discs to retain their discoid shape and viscoelastic function, containing vertical forces which may threaten spinal architecture leading to vertebral body compression fractures.
OBJECT: Given the virtual absence of histologically detected, clinically unsuspected disease in intervertebral disc specimens, some authors have advocated that histological examination be discontinued. However, the examination of intervertebral disc specimens remains common practice in most pathology laboratories. No cost-benefit analysis of this practice has been made; therefore, the authors' goal in this study was perform such an analysis. METHODS: Using the University of Iowa surgical pathology database, 1109 patients who had undergone a laminectomy were identified retrospectively. These cases were classified into four categories based on the patients' preoperative clinical diagnosis and final histopathological diagnosis: insignificant clinical diagnosis/insignificant pathological diagnosis (ICIP), significant clinical diagnosis/insignificant pathological diagnosis (SCIP), significant clinical diagnosis/significant pathological diagnosis (SCSP), and insignificant clinical diagnosis/significant pathological diagnosis (ICSP). A significant clinical diagnosis was defined as one other than a benign, noninfectious indication for laminectomy. A significant pathological diagnosis was a diagnosis other than degenerative changes. The cost-benefit value of performing a histological examination in cases with significant or insignificant clinical diagnoses was examined. The cases were classified as: 1068 ICIP, 17 SCIP, 21 SCSP, and three ICSP. On chart review, in all three cases of ICSP an epidural abscess was identified perioperatively and the subsequent histological diagnosis did not affect patient care. The costs per case of identifying a significant pathological diagnosis with a significant and an insignificant clinical diagnosis were $44.79 and $8811, respectively. CONCLUSIONS: Histological examination of intervertebral disc specimens is cost beneficial only if there is a significant preoperative clinical diagnosis.
OBJECTIVE: To test the hypothesis that hydrostatic pressure directly affects the synthesis of collagen and proteoglycan by intervertebral disc cells. DESIGN: By the use of pressure vessels, hydrostatic pressure was applied to intervertebral disc cells cultured in alginate. BACKGROUND: The influence of compression (both hydrostatic and axial) on chondrocyte metabolism was examined in a number of earlier studies. However, in most of these studies, articular cartilage, not intervertebral disc was used, and in none of these was hydrostatic pressure applied to intervertebral disc cells cultured in alginate. METHODS: Fresh cells were harvested from the lumbar intervertebral discs of dogs. Before their suspension in an alginate gel system, the cells were plated and expanded until they reached confluence. Then, by use of the alginate gel system, the cells were exposed (for up to 9 days) to specific values of hydrostatic pressure inside two stainless steel pressure vessels. One vessel was kept at 0.35 MPa and the other at atmospheric pressure (approximately 0.1 MPa). The effects of 0.35 MPa were compared against atmospheric pressure by measuring the incorporation of [3H]-proline and [35S]-sulfate into collagen and proteoglycans, respectively, for the anulus cells and nucleus cells separately, and by determining whether this incorporation was reflected by changes in the levels of mRNA for aggrecan and Types I and II collagen. RESULTS: Proteoglycan synthesis was inhibited at 0.35 MPa as compared to atmospheric pressure for both the nucleus and anulus cells, whereas collagen synthesis was stimulated in the nucleus cells, but inhibited in the anulus cells. The mRNA levels of collagen 1A and collagen 2A decreased in the anulus but showed a differential response in the nucleus (collagen 1A increased, while collagen 2A decreased). The mRNA levels for aggrecan core protein decreased in the anulus and increased in the nucleus. CONCLUSIONS: Hydrostatic pressure directly affects the synthesis of collagen and proteoglycan by the intervertebral disc cells. RELEVANCE: This in vitro study reveals the direct effect of hydrostatic pressure on disc cells, in the absence of other factors. However, circumspection must be applied when comparisons between these results, from in vitro experiments on dog disc cells, are extrapolated and applied to the whole discs of humans.
Surgical treatments for intervertebral disc disease are useful for animals with gradually progressive, severely acute, or recurrent lesions. Surgical patients require a diligent neuroradiographic evaluation to confirm location and severity of the discopathy and rule out conditions unrelated to disc disease. Although controversial, some surgeons advocate fenestration for recurrent pain or mild ataxia associated with a noncompressive lesion. Decompressive procedures are recommended for treatment and prognostication of compressive disc-associated myelopathy. Foraminotomy is reserved for patients with lateralizing extrusions confirmed with myelography or newer imaging modalities (CT, MRI). Future treatments for animals may include chemonucleolysis or various discectomy procedures currently performed in humans with symptomatic disc disease.
STUDY DESIGN: Immunohistochemical study of expression and localization of bone morphogenetic protein (BMP)-2/4 and type I and II receptors on intervertebral disc. OBJECTIVES: To determine the biologic functions of BMPs and their receptors in the process of degeneration of the intervertebral disc. SUMMARY OF BACKGROUND DATA: Biologic and pathologic processes in the cell during the degeneration of the intervertebral disc are as yet poorly understood. METHODS: The cervical spines of 15 male senescence-accelerated mice aged 8, 24, or 50 weeks were used for histologic and immunohistochemical examination of BMP-2/4 and BMP receptors IA, IB, and II. Immunostaining was performed with the avidin-biotin-peroxidase complex method. RESULTS: Degenerative change was recognized within intervertebral discs of senescence-accelerated mice aged 50 weeks. BMP-2/4 and its receptors were abundant in hyaline cartilaginous cells within the endplate of the vertebrae at 8 and 24 weeks of age. However, the expression of BMP-2/4 and its receptors moved from the hyaline cartilage of the endplate of the vertebrae to fibrous cells within the anulus and to the calcified cartilage at the site of enthesis of mice aged 50 weeks. CONCLUSIONS: BMP-2/4 and its receptors may play roles in degenerative change of intervertebral disc.
STUDY DESIGN: Immunohistochemical study of the sensory innervation of the human lumbar intervertebral disc. OBJECTIVE: To determine the type of sensory fibers innervating human degenerated lumbar intervertebral discs. SUMMARY OF BACKGROUND DATA: Sensory neurons involved in pain perception related to inflammation in rats are typically small, peptide-containing neurons immunoreactive for calcitonin gene-related peptide (CGRP). Small non-peptide-containing neurons binding to isolectin B4 (IB4) may also be involved in pain states, such as nerve injury pain. The character of such sensory neurons in humans has not been clarified. METHODS: A degenerated, painful lumbar intervertebral disc was harvested from each of 8 patients during surgery. Sections were immunostained for protein gene product 9.5 (PGP 9.5, a general neuronal marker), CGRP, and IB4. The numbers of PGP 9.5- and CGRP-immunoreactive, and IB4-binding nerve fibers in the discs were counted. RESULTS: PGP 9.5-immunoreactive fibers were observed in all discs. Nerve fibers immunoreactive for CGRP were also observed in 6 of 8 cases. IB4-binding nerve fibers were not found in any case. CONCLUSIONS: Almost all of the nociceptive nerve fibers in the human intervertebral disc are peptide-containing nerve fibers, similar to the rat disc, suggesting that nerve fibers related to inflammation may transmit pain originating from human degenerated intervertebral discs.
STUDY DESIGN: A case of intraradicular, intervertebral disc herniation is reported. OBJECTIVES: A rupture of en L5-S1 intervertebral disc into the left S1 nerve root is described. SUMMARY OF BACKGROUND DATA: A review of the literature revealed five reported cases with some possible causes of this extremely rare phenomenon. METHODS: The diagnosis was made at the time of surgery during exploration of the L5-S1 disc space. The S1 nerve root was found to be adherent to the underlying disc space. Before surgery, this condition was believed to represent a large extradural and extraradicular disc herniation. RESULTS: The patient's pain was alleviated almost immediately after surgery, but the patient was left with some residual sensory loss in the leg. CONCLUSION: An unusual case of intraradicular, intervertebral disc herniation is presented.
1H-nuclear magnetic resonance studies, in vitro, on normal intervertebral discs dissected from adult bovine (3- or 4-year-old) coccygeal vertebrae were performed to clarify topographic differences in relaxation times (T1, T2) and relationships between the relaxation times and water content. Immediately after death, intervertebral discs were dissected from 1st/2nd and 2nd/3rd coccygeal vertebrae and divided into three specimens: the nucleus pulposus, inner anulus fibrosus, and outer anulus fibrosus. T1, T2, and water content were measured for the specimens collected from 15 intervertebral discs (untreated group). Secondly, specimens were similarly collected from 12 intervertebral discs and were treated to changed water contents (dehydrated group and hydrated group). For each treated specimen, T1, T2, and water content were measured. The results showed highly significant differences (P < 0.01) in T1 values among the nucleus pulposus, inner anulus fibrosus, and outer anulus fibrosus, and also in T2 values between the nucleus pulposus and anulus fibrosus. In each of the nucleus pulposus and anulus fibrosus, a positive linear relationship (r = 0.97, P < 0.01) was found between T1 and water content. The respective regression lines for the nucleus pulposus and anulus fibrosus had different slopes, and the slope of the nucleus pulposus was significantly greater than that of the anulus fibrosus. Thus, the decrease in water content shortens the T1 in intervertebral discs, and changes in water content have a greater influence on T1 in the nucleus pulposus than in the anulus fibrosus.(ABSTRACT TRUNCATED AT 250 WORDS)
The collagen and proteoglycan contents were determined in lumbar intervertebral disc nucleus from 50 cases normal Chinese with accidental death and in that of 50 cases of prolapsed lumbar intervertebral disc (PID). The significance of the work rested with. (1) The normal range of the collagen and proteoglycan contents in the nucleus pulposus of normal Chinese was defined, therefore, it provided a normal parameter and laid a foundation for the study of collagen and proteoglycan metabolic changes in the nucleus pulposus of lumbar intervertebral disc under pathological condition. (2) By comparison of contents between collagen and proteoglycan in the nucleus pulposus of PID, the physiopathology of PID was confirmed further, that is, the content of proteoglycan of the nucleus pulposus in PID decreased, however, that of collagen increased. (3) The collagen contents between fibrocartilage and nucleus pulposus in normal chinese were compared. Analyses were made biomechanically and biochemically.
OBJECTIVE: To investigate the expression of integrin alpha5 and actin in the cells of intervertebral disc under cyclic hydrostatic pressure in vitro. METHODS: The porcine lumbar intervertebral disc cells were isolated and cultured in vitro, and the cells underwent cyclic hydrostatic loading. After that, the expression of integrin alpha5 and actin in intervertebral disc cells were studied by means of morphology observing, Western blot and immunohistochemistry staining. RESULTS: The morphology of intervertebral disc cells were changed into smaller and flatten shape, and the expression of integrin alpha5 and actin were decreased after loading. CONCLUSIONS: The expression of integrin alpha5 decreases under cyclic hydrostatic pressure, and the actin is affected at the same time when signals are transferred into the cells by integrin alpha5. That may be one of the important mechanisms of the mechanotransduction in the cells of intervertebral disc.
Immature bovine cartilages and intervertebral-disc tissue all revealed a prominent protein, not present in the adult tissues, in non-denaturing extracts made with chondroitin ABC lyase (EC 4.2.2.4), Streptomyces hyaluronidase (EC 4.2.2.1) or 1 M NaCl. The protein ran on SDS-polyacrylamide electrophoresis, before disulphide reduction, as a close doublet of bands of apparent molecular weight 110,000 and 105,000. After reduction, they dissociated respectively into two protein bands at 37,000 and 35,000, indicating that the initial molecules were disulphide-bonded trimers. Amino-terminal sequence analysis established the identity of both proteins (Mr 110,000 and Mr 105,000) as forms of the carboxypropeptide of type II collagen. The larger molecule appeared to be the trimer of intact alpha 1(II) carboxypropeptides and the smaller, a version composed of chains that were ten residues shorter at their amino-terminal ends. The material appears to be identical to chondrocalcin, a protein previously found to be enriched in fetal growth plate and named on the basis that it may play a role in cartilage calcification. The present findings, however, indicate that the protein is equally abundant in all type II collagen-synthesizing young cartilages, including nucleus pulposus of the intervertebral disc and other cartilages that never calcify.
Degeneration of the intervertebral disc is a common disease in the adults, especially at advanced age. A causal therapy is not known, but the progress in new therapeutic strategies, for example in tissue engineering, shows new possibilities. The goal of our study was to develop a new animal model that stimulates a load induced degeneration of the disc. We used the New Zealand rabbit, because morphology is similar to the human intervertebral disc. The degeneration was induced by axial compression of the disc L4 - L5 with an external fixateur. After different loading intervals, the animals were sacrified and the discs examined by radiology, histology, apoptosis and biomechanical testing. Radiography showed a significant decrease of the disc thickness in all loaded groups. Morphologically the intervertebral discs of loaded rabbits showed degenerative changes which were comparable to those in humans. A significantly increased number of dead cells in the annulus occurred after 14 and 28 days loading compared to the controls. The bending stress measured as the load to failure was not significantly different between the unloaded discs and the 28 days loaded discs. The results show that our animal modell can create degeneration. Four weeks compression leads to significant degeneration. Degeneration of the discs persisted in animals that were allowed a recovery time of 28 days after 28 days of loading.
In 64 surgically treated herniated lumbar intervertebral discs, we performed histopathological studies of neovascularization in the outermost layer of the herniated mass in various types of hernia. We obtained specimens separately from the capsule tissue covering the herniated mass and the inner tip tissue of the herniated mass for comparison. Histologically, in most cases, the capsule tissue was the outermost layer of the annulus fibrosus or posterior longitudinal ligament, and the inner tip tissue was the nucleus pulposus. In the capsule tissue, newly formed small blood vessels were present in 73.4% of the total cases examined, regardless of the hernia type. However, the frequency and degree of such vessels in the tip tissue were significantly higher in hernias that perforated the posterior longitudinal ligament than in those that did not. When the intervertebral disc herniates, new blood vessels proliferate in the capsule of the hernial tissue. At the stage when the hernial capsule tissue is still present, these vessels were observed to have difficulty reaching the inner tip portion. These findings suggest that when the nucleus pulposus portion of the herniated mass perforates the posterior longitudinal ligament, it may be subject to a stronger neovascularization reaction.
STUDY DESIGN: Various amounts of static mechanical load were applied to mouse intervertebral discs in organ cultures. The apoptosis then was examined using nick end labeling. Two mitogen-activated protein kinase (MAPK) inhibitors were added to the medium. OBJECTIVES: To establish an experimental model for detecting factors regulating chondrocyte apoptosis induced by mechanical stress, and to determine the role of MAPK and p38 in the stress-induced apoptotic pathway of endplate chondrocytes. SUMMARY OF BACKGROUND DATA: The cause of degenerative change in the cartilaginous endplate (CEP) remains unclear. The authors' previous findings using a mouse model suggested that apoptosis in the cartilaginous endplate may play a role in intervertebral disc degeneration, and that mechanical stress may induce apoptosis. If apoptosis of endplate chondrocytes is involved in the cascade of intervertebral disc degeneration, then how apoptosis is induced by mechanical stress should be important in preventing disc degeneration. However, the mechanism of apoptosis induced by mechanical stress remains unclear. METHODS: Mouse coccygeal discs were harvested and organ cultured. Various static compression loads (0, 0.2, 0.4, 0.8, and 1.0 MPa) were applied on intervertebral discs placed in culture bottles for 24 hours. Paraffin-embedded sections of the harvested discs were stained using Safranin-O and the nick end labeling procedure. The apoptotic cells were counted in the cartilaginous endplate and junctional anulus fibrosus of each intervertebral disc. In addition, U0126 (MAPK inhibitor) and SB202190 (p38 inhibitor) were added to the culture medium to determine their regulatory roles in the apoptosis of endplate chondrocytes induced by mechanical load. RESULTS: Histologically, loaded discs became bulged, and the disc space became narrow. Apoptosis was absent in discs without load, but was particularly noticeable in loaded discs (load weight, 1.0 MPa). The number of apoptotic cells increased depending on the weight of the load. The two MAPK inhibitors significantly increased the number of apoptotic cells. CONCLUSIONS: Chondrocyte apoptosis was induced using a static mechanical load especially in the cartilaginous endplate in an organ culture. Apoptosis occurred similarly to previous findings using an in vivo model. This culture system thus reflected the apoptosis demonstrated in vivo. Because biologically active reagents such as MAPK inhibitors can be simply added to culture media, this system may be a useful method for detecting factors that influence apoptosis induced by mechanical stress. Both MAPK inhibitors increased the occurrence of apoptosis. This suggests that these two MAPKs can counteract the apoptotic pathway induced by mechanical stress.