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PubMed2006

Intradiscal injections of osteogenic protein-1 restore the viscoelastic properties of degenerated intervertebral discs.

BACKGROUND CONTEXT: Using biochemical, histological, and radiological parameters in a rabbit model of intervertebral disc (IVD) degeneration, the intradiscal injection of a growth factor, such as osteogenic protein-1 (OP-1), has been shown to regenerate the IVD. However, very little is known about how such a biological therapeutic approach affects the biomechanical properties of the degenerated IVD. PURPOSE: To investigate the effects of an intradiscal injection of OP-1 on the biomechanical properties of IVDs in the rabbit annular-puncture disc degeneration model and to determine their relationship to biochemical properties. STUDY DESIGN/SETTING: In vivo study on the effects of intradiscally administered OP-1 on the biomechanical and biochemical properties of IVDs in the rabbit annular-puncture disc degeneration model. METHODS: New Zealand White rabbits (n=16) underwent annulus fibrosus (AF) puncture, using an 18-gauge needle, at L2-L3 and L4-L5 (L3-L4: nonpunctured control). Four weeks later, the punctured discs received an injection of either 5% lactose (10 microL) or OP-1 (100 microg/10 microL of 5% lactose) into the nucleus pulposus (NP). The disc height was radiographically monitored biweekly. After sacrifice and removal of bone-disc-bone complexes 8 weeks postinjection, the dynamic viscoelastic properties of the IVDs were tested by applying a cycle of sinusoidal strain in uniaxial compression at six loading frequencies (0.05 to 2 Hz). The biochemical properties of the dissected IVDs were then analyzed and correlated with the biomechanical properties. RESULTS: A single injection of OP-1 significantly restored disc height when compared with the lactose-injected discs (OP-1 vs. lactose, p<.001). The elastic modulus of the IVDs in the OP-1-injected discs was significantly higher than that in the lactose-injected discs at all frequencies (mean: +43%, p<.001). The viscous modulus in the OP-1-injected discs was significantly higher at 0.05, 0.2, 0.5, and 1 Hz (mean: +55%, p<.001) and showed higher tendencies at other frequencies (p=.08-.09). For both moduli, no significant differences were observed between the OP-1-injected and the nonpunctured control discs. The OP-1 injection significantly increased the proteoglycan (PG) content in the NP and AF, and the collagen content in the NP (p<.001-.05). Both elastic and viscous moduli showed significant positive correlations with PG content in the NP and collagen content in the NP and AF (Rho=.357-.466, p=.010-.047). CONCLUSIONS: We have shown for the first time that an injection of the growth factor, OP-1, restored the biomechanical properties of IVDs in a rabbit model of IVD degeneration. Comparing biomechanical with biochemical data suggests that the OP-1-induced biomechanical restoration was a consequence of increased activities of anabolic pathways that resulted in biochemical changes in the IVD.

Animals↗
PubMed2005

Osmolarity regulates gene expression in intervertebral disc cells determined by gene array and real-time quantitative RT-PCR.

Intervertebral disc (IVD) cells experience a broad range of physicochemical stimuli under physiologic conditions, including alterations in their osmotic environment. Cellular responses to altered osmolarity have been documented at the transcriptional and post-translational level, but mainly for extracellular matrix proteins. In this study, the gene expression profile of human IVD cells was quantified with gene array technology following exposure to increased osmolarity in order to capture the biological responses for a broad set of targets. A total of 42 genes were identified in IVD cells as significantly changed following culture under hyper-osmotic conditions. Gene expression patterns were verified using RT-PCR. Genes identified in this study include those related to cytoskeleton remodeling and stabilization (ephrin-B2, muskelin), as well as membrane transport (ion transporter SLC21A12, osmolyte transporter SLC5A3, monocarboxylic acid SLC16A6). An unexpected finding was the differential regulation of the gene for the neurotrophin, brain-derived neurotrophic factor, by hyper-osmotic stimuli that suggests a capability of IVD cells to respond to physicochemical stimuli with factors that may regulate discogenic pain.

Cells, Cultured↗
PubMed1994

Proteoglycan heterogeneity in the normal adult ovine intervertebral disc.

Proteoglycans (PGs) were isolated from 4 M GuHCl extracts of young adult ovine Intervertebral disc (IVD) tissues using sequential CsCl density gradient centrifugation, and a combination of gel-permeation and hydrophobic chromatography. A total of six PG sub-populations were identified in both the Annulus fibrosus (AF) and Nucleus pulposus (NP), i.e. two high buoyant density aggregatable PGs, two high buoyant density non-aggregatable PGs and two small, low-intermediate buoyant density, non-aggregatable, DS-rich PG species. These latter PGs were identified as biglycan and decorin on the basis of analyses of their core protein native size, and glycosaminoglycan composition. Additional low-intermediate buoyant density PG species were also evident in the non-aggregatable PG pool, particularly in extracts of NP tissues. These PGs did not bind to the octyl affinity matrix under the experimental conditions employed and thus were readily separated from the DS-PGs by hydrophobic chromatography, their constituent glycosaminoglycans (CS and KS) also differed and were of a smaller size to the CS and KS chains isolated from the large high buoyant density PGs, the small PGs which did not bind to octyl-sepharose may therefore represent distinct PG species in their own right. Differences were evident in the absolute size, and in the distribution of individual PG species in the respective IVD tissues but the NP always contained a larger proportion of high buoyant density non-aggregatable PGs of somewhat smaller size than those isolated from the AF. Decorin and biglycan, however, were generally more abundant in AF tissues.

Animals↗
PubMed1986

Quantitative analysis of intervertebral disc structure.

A reduction in signal intensity from the nucleus can be a feature of intervertebral disc disease. It has been established that T1 and T2 relaxation times of the nucleus decrease with age and that evidence of disc degeneration can be determined before classical clinical features are apparent. A robust multiple point algorithm has been developed which now enables proton density and T1 and T2 values to be computed for each pixel, providing images which are effectively spatial maps of these parameters. Such high resolution maps have provided quantitative data from volunteers, patients and cadavers. These studies have been carried out with specially constructed coils and the cadaveric information compared with cut sections. The spatial maps revealed detailed anatomic structures including the laminae of the annulus and relative levels of hydration. These levels, which are known to be related to the ability of the disc to withstand compressive loads, can now be measured in vivo. Analysis of the proton density and relaxation times in vivo has demonstrated that both water content and the chemical environment in the nucleus change during aging. The results are consistent with changes in the glycosaminoglycan content and fixed charged density measured by other chemical and physical techniques. Such detailed methods can be used to investigate the effects of aging and disease on disc structure and have enabled observations to be made of the effects of stress on the normal disc.

Adolescent↗
PubMed1986

1H nuclear magnetic resonance study of intervertebral discs. A preliminary report.

In vitro 1H pulse nuclear magnetic resonance (NMR) studies on human intervertebral disc tissues were performed to establish the characteristics of relaxation processes. The spin-lattice relaxation time, bound water and dry matter content were measured. Biopsy material from fresh cadavers was taken from different discs along the spinal column and from different sectors of some single discs. It is suggested that the measured parameters correlate with the biomechanical behavior of discs, which in turn is influenced by function, by exposure to physical stresses, or by aging.

Adolescent↗
PubMedDate not supplied

Molecular basis of intervertebral disc degeneration.

This review will acquaint the reader with normal development, structure, and function of the intervertebral disc. The disc is composed of the vertebral endplate, nucleus pulposus, and annulus fibrosus. These three functional components all serve important purposes for health and function of the disc. Nutrition and biomechanics are also discussed. The molecular basis of disc degeneration is reviewed so that biologic approaches to the reversal and or treatment of disc degeneration may be better understood.

Biomechanical Phenomena↗
PubMed1975

The proteoglycans of the human intervertebral disc.

The methods of Hascall & Sajdera (1969) were used to compare the proteoglycans of human intervertebral disc with those of bovine nasal cartilage. In contrast with cartilage, most of the hexuronate of disc could be extracted at low shear with water or dilute salt solutions. Extracts of disc with 4M-guanidinium chloride were centrifugated in 0.4M-guanidinium chloride in a CsCl gradient. Analytical ultracentrifugation of the hexuronate-containing heavy component revealed two fractions. both more polydisperse than those of cartilage. Also the more rapidly sediminting component was a much smaller fraction of the total. After prior extraction with 0.4M-guanidinium chloride, 4M-guanidinium chloride extracts of disc were found, by ultracentrifugal analysis, to be enriched in components resembling the proteoglycan monomer and aggregating factors of cartilage.

Animals↗
PubMedDate not supplied

Relation between the structure of the annulus fibrosus and the function and failure of the intervertebral disc.

A simple model is presented that explains the observed function and failure of the intervertebral disc in compression, torsion, and bending; this model is based upon the observed arrangement of collagenous fibers in the annulus fibrosus. The fibers are considered to have the same mechanical properties as tendon; thus the stresses required to produce a given deformation and which irreversibly damage the fibers can be predicted. Predictions of the mechanical behavior of the disc are in good agreement with published results for compression and torsion; no comparable experiments have been performed for bending. It is further predicted that torsion and bending are likely to cause annular failure and protrusion. Failure is likely to occur posteriorly because of the effect of forward bending and because in the flattened and reentrant discs of the lumbar spine, torsional stress is concentrated at the points of maximum curvature. The structure of the disc tends to protect the collagenous fibers in forward bending and torsion. Compression is predicted to cause end-plate fracture rather than annular failure.

Back Pain↗
PubMed1996

Pathomechanism of pain-related behavior produced by allografts of intervertebral disc in the rat.

STUDY DESIGN: This study was designed to evaluate whether allografts of intervertebral disc materials produce hyperalgesia in the rat and whether an immune response, pH, or chemicals correlate with the induced hyperalgesia. OBJECTIVE: To elucidate the pathomechanisms of radicular pain secondary to lumbar disc herniation. SUMMARY OF BACKGROUND DATA: It has been reported that a low pH, an autoimmune reaction, or chemical radiculitis is likely responsible for radicular pain associated with lumbar disc herniation. In animal studies, it has been shown that hyperalgesia (an increased sensitivity to painful stimuli) involves activation of phospholipase A2 and nitric oxide synthase. METHODS: Fat, nucleus pulposus, and anulus fibrosus were allografted into the epidural space at L6 in the rat. Withdrawal response thresholds to mechanical stimuli and withdrawal response latencies to thermal stimuli on the tail and pH in the applied tissues were measured after surgery. Interleukin-1, phospholipase A2, and nitric oxide synthase were examined in the applied tissues using immunohistochemistry, nicotineamide adenine dinucleotide phosphate-diaphorase histochemistry, and in situ hybridization. RESULTS: Allografted fat did not produce hyperalgesia. Allografts of nucleus pulposus and nucleus pulposus plus anulus fibrosis showed evidence of mechanical and thermal hyperalgesia, respectively. There were no observed changes in pH over time. Although interleukin-1 was demonstrated in all applied tissues, phospholipase A2 was only observed around the applied nucleus A2 was only observed around the applied nucleus pulposus and nucleus pulposus plus anulus fibrosus. Nitric oxide synthase was only markedly increased around the applied tissues. CONCLUSION: The nucleus pulposus and anulus fibrosus produce different forms of hyperalgesia (mechanical vs. thermal) associated with different and distinct immunohistochemical changes. It is possible that radicular pain of a lumbar disc herniation results from chemicals, such as phospholipase A2 and nitric oxide.

Adipose Tissue↗
PubMed1999

[Anamnestic and anthropometric assessment of patients with herniated lumbar intervertebral disc].

The paper presents results of anamnestic and anthropometric assessment of patients with herniated lumbar intervertebral disc operated in the Department in the years 1978-1987 (983 patients) and in the decade 1988-1997 (1308 patients). Recent decade brought 33.1% increase in disectomies. Typically, discopathy patient is tall individual, with relatively high BMI and aged between 19 and 48 years. The number of female patients increased by 8.2% in recent decade and traumatic origin of the disc herniation was noted in 8.8% more than previously. Special attention should be paid to the twofold increase of female smokers among patients operated due to discopathy.

Adult↗