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At least 1,171 records · Page 65Linked to original sources
PubMed2006

In vitro organ culture of the bovine intervertebral disc: effects of vertebral endplate and potential for mechanobiology studies.

STUDY DESIGN: Whole bovine coccygeal discs were cultured under static load, with or without vertebral endplates (VEPs), and assessed for cell viability, biochemical stability, biosynthetic activity, and biosynthetic responsiveness to changes in mechanical load. OBJECTIVES: To assess the effects of VEPs on biochemical and cellular stability of disc cells during in vitro culture of large disc explants. To determine whether cultured discs could respond to mechanical perturbation. SUMMARY OF BACKGROUND DATA: Previous methods for culturing the intervertebral disc have focused on rabbit and rat discs, but the small size of these discs limits the relevance of these culture systems to the human condition. Bovine coccygeal discs have similar dimensions to the human lumbar disc (i.e., similar size and nominal stresses), but long-term culture of these discs has not been reported. METHODS: Bovine coccygeal discs were harvested with or without VEPs, cultured under static load (5 kg, approximately 0.25 MPa, in situ swelling pressure) for up to 1 week, and evaluated for changes in hydration, glycosaminoglycan content, cell viability, and biosynthetic activity. Additionally, the biochemical and biosynthetic response of discs cultured without VEP to increasing the load to a 20-kg (approximately 1 MPa, the estimated stress in human lumbar disc during heavy lifting) static load for 6 hours was assessed. RESULTS: During the first 24 hours, culturing discs with endplates was moderately better with regards to maintaining in situ anulus hydration and nucleus glycosaminoglycan levels. The endplates, however, obstructed media flow to the disc, resulting in a marked decrease in cell viability after 1 week of culture. Nucleus pulposus cell viability was maintained in discs cultured without endplates, but there was a significant drop in biosynthetic activity within 2 days of culture. Despite this drop, the disc cells in the discs without VEP remained biosynthetically responsive to changes in mechanical loading. CONCLUSIONS: It is possible to maintain cell viability and the biosynthetic responsiveness of large discs for up to 1 week in vitro when the discs are cultured under static load and without VEP.

Animals↗
PubMed1986

The effects of chymopapain on prolapsed human intervertebral disc. A clinical and correlative histochemical study.

Light and electron microscopy were used to demonstrate extensive matrix degradation in a failed chymopapain-treated disc as compared with an untreated degenerative control disc. Both specimens contained viable cells with patches of degenerative cells. There was a loss of disc height in the chymopapain-treated disc, but no improvement of symptoms. These results illustrated that even in well-circumscribed cases with documented disc protrusion into the spinal canal, the disc space narrowing following injection may aggravate the symptoms of nerve root entrapment, especially if a low-grade lateral recess stenosis is associated with the disc protrusion. The persistence or aggravation of symptoms probably is related to these anatomic considerations. The low level of enzyme activity in the injected area may or may not be correlated with the clinical response to treatment.

Aged↗
PubMed1977

The variation of the glycosaminoglycans of the canine intervertebral disc with ageing. I. Chondrodystrophoid breed.

The variation with age and spinal level of the glycosaminoglycan distribution in beagle (a chondrodystrophoid breed) intervertebral discs have been assessed by means of histochemical and chemical methods. Chondroitin sulphate is the major glycosaminoglycan of the immature beagle disc, but with ageing, this is partially replaced by keratosulphate. The level of glycosaminoglycans in the nucleus pulposus is markedly less than in the nonchondrodystrophoid breed and by 1 year of age, the beagle nucleus pulposus has become largely fibrocartilagious. This alteration in 'normal' disc developments is possibly associated with the elevated incidence of disc disorders in the chondrodystrophoid breeds.

Aging↗
PubMed1998

Analysis of aging and degeneration of the human intervertebral disc. Comparison of surgical specimens with normal controls.

STUDY DESIGN: A prospective analysis of 33 control and 39 surgical human lumbar disc specimens from the anulus was undertaken to assess disc cell extracellular matrix production and cell function. The authors of this study analyzed immunohistochemical distributions of Types I, II, III and VI collagen, in situ localization of apoptotic disc cells, and tartrate-resistant acid phosphatase localization. OBJECTIVES: To quantify the incidence of apoptotic cell death in the anulus; examine the collagen distribution in the pericellular, territorial, and interterritorial matrix; examine matrix cell degeneration; and compare diseased tissue with normal tissue from control individuals. SUMMARY OF THE BACKGROUND DATA: Previous studies of disc histopathology have focused on extracellular matrix morphology and on biochemical synthetic and degenerative changes, but little is understood about the cell populations within the disc that are responsible for these changes. METHODS: In this study light microscopy, immunohistochemistry, enzyme histochemistry, and in situ hybridization were used to examine 33 patient and 39 control specimens of human anulus obtained either via surgical procedures or from donors to the Cooperative Human Tissue Network. RESULTS: The high incidence of apoptotic cell death was significantly greater in the control group (73.1 +/- 5.1% [mean +/- SEM]; n = 20) than among surgical specimens (53.5 +/- 5.6%; n = 20; P < 0.001); this was probably a result of the significantly greater average age in the control population (57.2 +/- 3.1 years) compared with that in the patient population (44.3 +/- 3.2 years; P < 0.001). Immunohistochemistry yielded findings that led to an expanded appreciation of the greatly modified extracellular domains that surrounded disc cells during aging and degeneration in both study groups. Enzyme histochemistry revealed the presence of tartrate-resistant acid phosphatase activity in human disc cells. CONCLUSIONS: These findings reveal that there is a high incidence of apoptosis in the intervertebral disc. Surviving cells are not synthetically inactive but are, rather, producing inappropriate matrix products during aging and degeneration. In certain instances it appears that the matrix surrounding the cell may form an isolation barrier, which may influence individual cell activity and intercellular communication. These results point to the need to 1) more fully understand the cause of disc cell death via apoptosis and to determine whether this programmed cell death can be reversed or halted, and 2) more fully understand the dynamic relation between disc cells and the surrounding extracellular matrix, which they produce and remodel. The factors regulating extracellular matrix-disc cell homeostasis in the disc are unknown, as is the relation between extracellular matrix and disc cell functional modulation. The morphologic findings of this study suggest that these issues are important considerations in disc cell biology. The identification of tartrate-resistant acid phosphatase activity in disc cells allows for a new area of study of disc extracellular matrix remodelling. In summary, these new perspectives provide new parameters with which to assess disc cell health and function.

Adult↗
PubMed2001

Regional variations in certain cellular characteristics in human lumbar intervertebral discs, including the presence of alpha-smooth muscle actin.

An evaluation of the regional variation of certain cellular features in the human intervertebral disc (IVD) could lead to a better understanding of site-specific properties relative to degradation, response to injury, and healing processes. The objective of this study was to determine how cell density, cell morphology, cell grouping, and expression of a specific actin isoform varied with location and degeneration in the human disc. A total of 41 human L4-L5 and L5-S1 discs removed postmortem from 21 individuals were analyzed. The discs were graded for degeneration based on the Thompson scale and processed for evaluation. Microtomed sections from paraffin-embedded specimens were stained with hematoxylin and eosin or a monoclonal antibody to alpha-smooth muscle actin (alpha-SMA), an actin isoform often associated with contraction. A significant regional dependence was found for most of the measured parameters. A fourfold increase in cell density was found in proceeding from the nucleus pulposus (NP) to the outer annulus (OA) of the IVD. Approximately 30% of the cells in the NP were present in groups. Virtually all of the cells in the NP and 40% of those in the OA were round. Moreover, notable percentages (12-15%) of the cells in the NP and inner annulus (IA) contained alpha-SMA. Only pair density was found to be correlated with Thompson grade, with more degenerated specimens having higher values. A greater effect was also observed on the percentage of cells in groups. These findings provide the basis for future work to investigate the importance of cells in groups, the role of alpha-SMA in the disc, and the changes in these cellular characteristics in pathological disc conditions.

Actins↗
PubMed2002

Autologous intervertebral disc cell implantation: a model using Psammomys obesus, the sand rat.

STUDY DESIGN: Work presented here used a small animal model to illustrate the feasibility of autologous disc cell implantation. OBJECTIVES: To develop a small animal model for autologous disc cell implantation. SUMMARY OF THE BACKGROUND DATA: The use of autologous disc cells in the potential treatment of disc degeneration offers attractive possibilities for novel therapies. Results are presented on the use of the sand rat (Psammomys obesus), a small rodent that spontaneously develops disc degeneration during aging, in experimental studies in which cells were harvested from a lumbar intervertebral disc, expanded in monolayer tissue culture, labeled with agents that allow subsequent immunolocalization of these cells, and implanted in a second disc site of the donor animal. METHODS: Tissue culture, disc surgery, histology, and immunocytochemistry were used. Cells were either engrafted in a bioresorbable carrier tested for cell compatibility or injected into the recipient disc. Results were assessed with radiographic examination of the implantation site and with histology and immunocytochemistry. CONCLUSION: Data from 15 animals were obtained with engraftment resident in the animal for up to 33 weeks. Immunocytologic identification of engrafted cells showed that they integrated into the disc and were surrounded by normal matrix at time points up to 8 months postengraftment. Engrafted cells exhibited either a spindle-shaped morphology in the annulus or a rounded chondrocyte-like morphology in the nucleus. Although technically challenging, the authors' experience showed that autologous disc cell implantation can be successful and that the sand rat is a valuable model for autologous disc cell studies.

Animals↗
PubMed1982

Carbohydrate metabolism and concentration profiles of solutes in the canine lumbar intervertebral disc.

Utilization of glucose and oxygen by the cells as well as concentration profiles of solutes were studied in the canine intervertebral disc. Cell metabolism in this avascular tissue, being predominantly anaerobic, was found to be dependent on oxygen tension (this being especially pronounced in the region of 0.13-0.4 kPa (1-3 mmHg). A high production rate of lactic acid was found in the nucleus pulposus, whereas towards the periphery of the annulus fibrosus this rate gradually decreased. In the centrally located areas of the disc tissue, far away from the blood circulation, the highest concentrations of lactic acid were found. For the normal disc the energy demands seem to be met as even small amounts of oxygen account for a large energy source, whereas the cellular requirements are balanced up predominantly by glucose. In regions with extremely low oxygen tensions large amounts of glucose are consumed, but an additional potential energy pool of glycogen seems to be available.

Animals↗
PubMed2003

The prevalence of subclinical gastroduodenal ulceration in Dachshunds with intervertebral disc prolapse.

Endoscopy was used to determine the prevalence of subclinical gastroduodenal ulceration in 30 Dachshunds undergoing decompressive surgery for acute intervertebral disc prolapse. The endoscopy was performed on the day of admission and on the 3rd or 4th day after surgery. Three regions of the stomach (cardia, corpus and pylorus) and the proximal duodenum were visually inspected and biopsy samples were taken for histopathology. The combination of visual and microscopic changes were then used to determine the prevalence of subclinical gastroduodenal ulceration in this population. An overall prevalence of 76% was calculated from these findings. Ulcerogenic medication administered prior to admission did not appear to influence the prevalence. This result identifies a need for veterinarians to be aware of this potentially severe complication and warrants the use of prophylactic anti-ulcer medication in spinal surgery patients.

Animals↗
PubMed1999

Laminectomy for 34 dogs with thoracolumbar intervertebral disc disease and loss of deep pain perception.

The case details and the results of treatment of 34 dogs with thoracolumbar intervertebral disc disease, without deep pain perception, that had been treated by laminectomy and fenestration, are presented. The association of a number of potential prognostic factors with the neurological outcome is examined. Twenty-one dogs (62 per cent) recovered neurological function, seven (21 per cent) failed to recover neurological function and three (9 per cent) developed progressive myelomalacia postoperatively, while three dogs (9 per cent) were euthanized intraoperatively because of diffuse myelomalacia. Twenty of the dogs that recovered neurological function showed a return of deep pain perception within two weeks of decompressive surgery. Statistical analysis showed significant differences in the outcome between dogs that took less than one hour to lose the ability to ambulate and dogs with a longer duration of onset of inability to ambulate. The extent of spinal cord swelling determined by myelography was not found to be a useful prognostic indicator.

Animals↗