chondroitin-sulfates and Intervertebral-Disc-Displacement

Whole ovine caudal intervertebral discs (IVD) were cultured in sufficient and limited nutrition under simulated-physiologic loading for 7 and 21 days.. To study the effect of limited nutrition on disc cells embedded in their native tissue in short- and midterm whole organ disc culture.. Nutrient-limited induction of disc cell death in vitro has been demonstrated and is believed to be a factor in disc degeneration. Nutrient-limited cell death and its consequences, as it relates to degeneration, have not been investigated in the intact IVD.. Ovine IVDs with endplates were cultured for 7 and 21 days under simulated-physiologic loading, either in media with limited (2 g/L) or sufficient (4.5 g/L) glucose concentration. Cell viability, relative gene expression, newly synthesized chondroitin sulfate content, and matrix metalloproteinase (MMP) activity were measured after culture and compared to fresh tissue.. In sufficient glucose media, cell viability was maintained through 7 days to 21 days of culture. In limited glucose, it dropped significantly to 62% in the anulus fibrosus and to 56% in the nucleus pulposus after 7 days and remained so until 21 days (63% in the anulus fibrosus and 52% in the nucleus pulposus). No significant differences were found between culture conditions for relative gene expression, newly synthesized chondroitin sulfate and inactive and active forms of MMP13 and MMP7.. With this culture system, whole IVD explants could be maintained up to 21 days. Cell viability decreased to 50% to 60% under limited nutrition within days and remained so up to 3 weeks. The surviving cells did not compensate matrix production in this time frame.

Topics: Animals; Cell Death; Cell Survival; Chondrocytes; Chondroitin Sulfates; Compressive Strength; Extracellular Matrix; Fibrocartilage; Gene Expression; Glucose; Intervertebral Disc; Intervertebral Disc Displacement; Ischemia; Matrix Metalloproteinases; Organ Culture Techniques; Sheep, Domestic; Weight-Bearing

The degeneration of intervertebral disc (IVD) is a major cause of low back pain. However, there is no satisfactory preventive treatment for degenerative disc disease (DDD). In this study, we examined the effects of a novel cross-linked hyaluronate hydrogel and cross-linked chondroitin sulfate (CS) hydrogel on a rabbit model of IVD injury. We injected 300 microl of phosphate buffer saline, 1% sodium hyaluronate, cross-linked hyaluronate hydrogel, or cross-linked CS hydrogel into the injured IVDs. One, three or six months after treatment, the whole spinal columns were dissected and magnetic resonance (MR) images of the IVDs were examined. It was noted that the IVD, which was injected with cross-linked hyaluronate hydrogel or cross-linked CS hydrogel mostly retained the normal signal intensity of the MR images. These IVDs exhibited a higher degree of staining with safranin-O than the control discs or 1% sodium hyaluronate-injected discs, suggesting that the intradiscal application of cross-linked hyaluronate hydrogel or cross-linked CS hydrogel probably inhibits the degenerative cascade of the DDD. The intradiscal administration of these drugs is safe, easy and costs less. In the near future, these intradiscal injections may become the standard therapy for the treatment of DDD instead of the spine surgeries.

Topics: Alginates; Animals; Chondroitin Sulfates; Cross-Linking Reagents; Disease Models, Animal; Humans; Hyaluronic Acid; Intervertebral Disc; Intervertebral Disc Displacement; Rabbits; Treatment Outcome

Glucosamine and chondroitin sulfate preparations are widely used as food supplements against osteoarthritis, but critics are skeptical about their efficacy, because of the lack of convincing clinical trials and a reasonable scientific rationale for the use of these nutraceuticals. Most trials were on osteoarthritis of the knee, while virtually no documentation exists on spinal disc degeneration. The purpose of this article is to highlight the potential of these food additives against cartilage degeneration in general, and against symptomatic spinal disc degeneration in particular, as is illustrated by a case report. The water content of the intervertebral disc is a reliable measure of its degeneration/ regeneration status, and can be objectively determined by Magnetic Resonance Imaging (MRI) signals.. Oral intake of glucosamine and chondroitin sulfate for two years associated with disk recovery (brightening of MRI signal) in a case of symptomatic spinal disc degeneration. We provide a biochemical explanation for the possible efficacy of these nutraceuticals. They are bioavailable to cartilage chondrocytes, may stimulate the biosynthesis and inhibit the breakdown of their extracellular matrix proteoglycans.. The case suggests that long-term glucosamine and chondroitin sulfate intake may counteract symptomatic spinal disc degeneration, particularly at an early stage. However, definite proof requires well-conducted clinical trials with these food supplements, in which disc de-/regeneration can be objectively determined by MRI. A number of biochemical reasons (that mechanistically need to be further resolved) explain why these agents may have cartilage structure- and symptom-modifying effects, suggesting their therapeutic efficacy against osteoarthritis in general.

Topics: Chondroitin Sulfates; Dietary Supplements; Drug Administration Schedule; Drug Combinations; Glucosamine; Humans; Intervertebral Disc; Intervertebral Disc Displacement; Lumbar Vertebrae; Magnetic Resonance Imaging; Male; Middle Aged; Range of Motion, Articular; Remission Induction; Water; Weight-Bearing

Chondroitinase ABC (C-ABC) is expected to be a novel agent for chemonucleolysis. The effect of C-ABC was investigated by magnetic resonance (MR) and radiograph. C-ABC was administered into the lumbar intervertebral disks on the clinically normal beagles (n=5), in a dose of 50 microl (12.5 units as C-ABC). MR scans were performed pre-dose, and 1, 3, 7, 14 and 28 days after administration of C-ABC, and the signal intensity (SI) of the nucleus pulposus was measured. Radiographs were taken pre-dose, and 1, 2, 3, 4, 5, 6, 7, 14 and 28 days post-dose, to evaluate narrowing of the disk space in terms of height index (HI). In addition, the quantity of the chondroitin sulfate (CS) and the hyaluronic acid (HA) in the nucleus pulposus were measured by high performance liquid chromatography on day 28 after dosing. SI and HI continuously decreased, following the injection to 37.1% and 78.9% of the pre-dose values, respectively. Statistically significant differences (p<0.01) were observed between the C-ABC group and the control group in the respects on day 1 post-dose. CS and HA contents of the nucleus pulposus were noted to be significantly decreased on day 28 (p<0.01) in the treated group. This agent proved to degenerate proteoglycans in the nucleus pulposus, thus progressively reducing the interdiskal pressure from day 1 post-dose onwards. It is concluded that C-ABC is expected to afford its efficacy from early in the course of chemonucleolysis.

Topics: Animals; Chondroitin ABC Lyase; Chondroitin Sulfates; Dog Diseases; Dogs; Hyaluronic Acid; Intervertebral Disc; Intervertebral Disc Chemolysis; Intervertebral Disc Displacement; Lumbar Vertebrae; Magnetic Resonance Imaging; Male; Radiography

Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of nondiabetic (n = 5) and diabetic (n = 5) age-matched subjects. The discs used were discarded material from operations.. To clarify the reason for the higher risk of disc prolapse in diabetic patients.. The pathogenesis of diabetes results from a combination of neurologic dysfunctions and a yet undefined metabolic failure, which leads to an abnormal proteoglycan profile.. The following methods were used to determine the proteoglycan profile: the measurement of 35S-sulfate uptake per gram wet tissue into sulfated glycosaminoglycan using fresh tissue explants; extraction of proteoglycans by 4 M guanidinium chloride containing protease inhibitors, with further purification by ultracentrifugation on cesium chloride buoyant density gradient under dissociative conditions; total uronic acid and protein contents in the various gradient fractions; assessing the length of sugar side chains of isolated 35Sulfate-glycosaminoglycan molecules by separation of the glycosaminoglycan molecules on a Sepharose 6B-CL column; and paper chromatography of the final digest products of glycosaminoglycan molecules obtained by chondroitinase ABC, a glycosaminoglycan-degrading enzyme.. The findings show that discs from normal nondiabetic subjects have 15 times the rate of 35Sulfate incorporation into glycosaminoglycan molecules than do discs of diabetic patients. The proteoglycans of diabetic patients are banded at a lower buoyant density, indicating a lowered glycosylation rate and a lower number of sugar side chains per core protein. In discs of diabetic patients, there is a slight increase in the chain length of chondroitin sulfate. Further analysis of the glycosaminoglycan chains showed a decreased amount of keratan sulfate, compared with that in nondiabetic subjects. However, the total uronic acid content of the disc tissues and the ratio of uronic acid to protein of each fraction were unchanged in diabetic patients versus that in control subjects.. Discs in patients with diabetes have proteoglycans with lower buoyant density and substantially undersulfated glycosaminoglycan, which with the specific neurologic damage in these patients, might lead to increased susceptibility to disc prolapse.

Topics: Aged; Chondroitin Sulfates; Chromatography, Gas; Diabetes Complications; Diabetes Mellitus; Female; Glycosaminoglycans; Humans; Intervertebral Disc; Intervertebral Disc Displacement; Keratan Sulfate; Low Back Pain; Lumbar Vertebrae; Male; Middle Aged; Proteoglycans; Uronic Acids

The human intervertebral discs which were obtained by cadavers and anterior discectomy are investigated histochemically. Chondroitin-4S, chondroitin-6S, dermatan sulfate, hyaluronic acid and keratan sulfate were detected in the human intervertebral disc by various histochemical methods. pH2.5, pH1.1 toluidin blue metachromasia and 0.4M MgCl2 alcianophilia became weaker with increasing age, and the herniated disc were weaker than controlled discs in the same age group. Chondroitin-4S and chondroitin-6S were distributed throughout the discs. There was no clear localization of the various glycosaminoglycans in the human intervertebral disc, with the exception of keratan sulfate. There was no histologically and histochemically observable difference in the cervical and lumbar discs.

Topics: Adult; Cadaver; Cartilage; Chondroitin Sulfates; Dermatan Sulfate; Histocytochemistry; Humans; Hyaluronic Acid; Intervertebral Disc; Intervertebral Disc Displacement; Keratan Sulfate