chondroitin-sulfates and Knee-Injuries

Osteoarthritis is a disabling disease of the aging generation, which results in loss of quality of life and increased healthcare costs. Cytokines appear to play an important role in the cartilaginous degeneration characterizing the pathological process. Increasing experience is being gained with cytokine-modulating therapies aimed at interfering with effects of chondrodegradative cytokines in the synovial fluid. Although in vitro and in vivo effectiveness of several of these therapies has been demonstrated, clinical effectiveness remains disputable, which may be related to the low levels of inflammatory cytokines found in osteoarthritic joints. By contrast, directly after joint trauma, which has been shown to predispose to early osteoarthritis, synovial fluid cytokine levels are strongly increased. Cytokine-modulating therapies, however, have hardly been considered for this indication. Increased knowledge of intra-articular soluble mediators correlating with cartilage pathology will lead to further development of cytokine-modulating products and, eventually, to effective inhibition of cartilage degeneration, in both the osteoarthritic as well as injured joints.

Topics: Animals; Cartilage; Chondroitin Sulfates; Cytokines; Humans; Immunotherapy; Inflammation; Knee Injuries; Knee Joint; Osteoarthritis; Risk Factors; Synovial Fluid

An understanding of diurnal change is one of the important milestones for either biomarker validation or therapeutic level monitoring. The present study determines the most suitable period during the day for serum chondroitin sulfate WF6 (CS-WF6) and hyaluronic acid (HA) collection, and identifies the possible factors which affect the estimated putative half-life of serum CS-WF6 and hyaluronic acid (HA).. Forty-nine volunteers were enrolled in the present study, 22 healthy, 14 with anterior cruciate ligament (ACL) injury, and 13 volunteers with osteoarthritis (OA). Blood sample collection was carried out every four hours starting at 18.00 hours for 24 hours, with additional samples taken at 07:00 and 08:00 hours. Serum CS-WF6, HA levels were determined by an ELISA-based assay.. The serum CS-WF6 level was significantly different between the normal and both pathological conditions. The serum HA level was significantly different in every condition. There was no diurnal pattern of serum CS-WF6 and HA during the 24 hour period. An estimated putative half-life of serum CS-WF6 and HA was 4.32 ± 2.63 and 4.10 ± 2.34, respectively. The maximum CS-WF6, creatinine clearance (CrCl) level and body mass index (BMI) were not related to the changes of the WF6 half-life. The higher maximum HA and CrCl level related to the longer half-life of serum HA level, p = 0.008 and p = 0.001, respectively.. There was no diurnal pattern of serum CS-WF6 and HA due to the present study approach. Two hours after awakening in official time would be the suitable for serum CS-WF6. Two hours after awakening and after meals were suitable times for serum HA collection.

Topics: Adult; Aged; Biomarkers; Case-Control Studies; Chondroitin Sulfates; Circadian Rhythm; Female; Humans; Hyaluronic Acid; Knee Injuries; Male; Osteoarthritis, Knee; Young Adult

Various methods have been tried to treat the main meniscus problem, meniscal tears, for which we believe tissue engineering could be a viable solution. In this study, a three dimensional, collagen-based meniscus substitute was prepared by tissue engineering using human fibrochondrocytes and a collagen based-scaffold. This construct was made with 3 different collagen-based foams interspaced with two electrospun nano/microfibrous mats. The top layer was made of collagen type I-chondroitin sulfate-hyaluronic acid (Coll-CS-HA), and the middle and the bottom layers were made of only collagen type I with different porosities and thus with different mechanical properties. The mats of aligned fibers were a blend of collagen type I and poly(L-lactic acid-co-glycolic acid) (PLGA). After seeding with human fibrochondrocytes, cell attachment, proliferation, and production of extracellular matrix and glucoseaminoglycan were studied. Cell seeding had a positive effect on the compressive properties of foams and the 3D construct. The 3D construct with all its 5 layers had better mechanical properties than the individual foams.

Topics: Animals; Biocompatible Materials; Biomechanical Phenomena; Cell Adhesion; Cell Proliferation; Cell Shape; Cells, Cultured; Chondrocytes; Chondroitin Sulfates; Collagen Type I; Extracellular Matrix; Humans; Hyaluronic Acid; Knee Injuries; Lactic Acid; Materials Testing; Menisci, Tibial; Molecular Conformation; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Tibial Meniscus Injuries; Tissue Engineering; Tissue Scaffolds; Wound Healing

Although it has been shown that aggrecanases are involved in aggrecan degradation, the role of MMP (matrix metalloproteinase) aggrecanolysis is less well studied. To investigate MMP proteolysis of human aggrecan, in the present study we used neoepitope antibodies against MMP cleavage sites and Western blot analysis to identify MMP-generated fragments in normal and OA (osteoarthritis/osteoarthritic) cartilage, and in normal, knee injury and OA and SF (synovial fluid) samples. MMP-3 in vitro digestion showed that aggrecan contains six MMP cleavage sites, in the IGD (interglobular domain), the KS (keratan sulfate) region, the border between the KS region and CS (chondroitin sulfate) region 1, the CS1 region, and the border between the CS2 and the G3 domain, and kinetic studies showed a specific order of digestion where the cleavage between CS2 and the G3 domain was the most preferred. In vivo studies showed that OA cartilage contained (per dry weight) 3.4-fold more MMP-generated FFGV fragments compared with normal cartilage, and although aggrecanase-generated SF-ARGS concentrations were increased 14-fold in OA and knee-injured patients compared with levels in knee-healthy reference subjects, the SF-FFGV concentrations did not notably change. The results of the present study suggest that MMPs are mainly involved in normal aggrecan turnover and might have a less-active role in aggrecan degradation during knee injury and OA.

Topics: ADAM Proteins; ADAMTS4 Protein; Adolescent; Adult; Aged; Aged, 80 and over; Aggrecans; Cartilage, Articular; Chondroitin Sulfates; Extracellular Matrix; Humans; Keratan Sulfate; Knee Injuries; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Middle Aged; Osteoarthritis; Peptide Fragments; Procollagen N-Endopeptidase; Protein Interaction Domains and Motifs; Proteolysis; Recombinant Proteins; Substrate Specificity; Synovial Fluid; Young Adult

To examine different aggrecanase generated fragments in synovial fluid (SF) from patients with acute and chronic knee injuries and from knee healthy subjects.. We prepared SF-D1 samples from acute (n=35) and chronic (n=35) knee injury patients and knee healthy subjects (n=10). Aggrecan fragments were analyzed in the SF-D1 samples by quantitative (G1, ARGS, KEEE and G3 antibodies) and non-quantitative (GRGT and AGEG antibodies) Western blot.. ARGS-SELE, ARGS-chondroitin sulfate (CS)1, GRGT-, GLGS- and AGEG-G3 fragments were the main ARGS and G3 fragments in injured and reference samples. In the acute injury samples the concentrations of these fragments were increased compared to the reference, and the level of the ARGS-SELE remained elevated for at least 2 years after the joint injury. Both SF ARGS fragments and aggrecanase generated G3 fragments had high sensitivity and specificity as biomarkers in distinguishing injured from healthy knee joints, although the ARGS fragments had higher area under the receiver operating characteristic curve (AUC) values for injuries (74-86%) than the G3 fragments (AUC values 63-68%).. Our results suggest that during the acute phase after knee injury there is an increased aggrecanase activity against both the interglobular domain (IGD) and the CS2 cleavage sites of joint cartilage aggrecan. This increase in SF aggrecanolytic fragments is present for several years after the injury. SF ARGS fragments are better biomarkers than the aggrecanase generated G3-fragments in distinguishing injured from healthy knee joints.

Topics: Adult; Aggrecans; Blotting, Western; Cartilage, Articular; Case-Control Studies; Chondroitin Sulfates; Endopeptidases; Female; Humans; Knee Injuries; Male; Middle Aged; Proteolysis; Sensitivity and Specificity; Synovial Fluid; Young Adult

Serum chondroitin sulfate epitope (WF6) and hyaluronic acid (HA) levels were determined to be of clinical relevance to an anterior cruciate ligament (ACL) injury. This cross-sectional study recruited participants from two distinct groups. Group A was comprised of 74 healthy controls, and group B consisted of 33 ACL injury patients. Serum samples were taken and assayed by a competitive immunoassay with monoclonal antibody WF6. Serum HA was also determined by an ELISA-based assay using biotinylated HA-binding proteins. Both groups A and B shared similar values of age, body mass index, white blood cell count and percentage of polymorphonuclear cells. ESR levels were also shown to be within normal limits. The serum WF6 epitope levels of group B were significantly higher than those of group A, whereas serum HA levels were not different between the two groups. The serum WF6 epitope level is more sensitive to changes in articular cartilage due to a non-inflammatory instability condition than the serum HA level, and should prove to be one of the most promising assays for early post-traumatic arthritis detection.

Topics: Adolescent; Adult; Anterior Cruciate Ligament Injuries; Biomarkers; Cartilage, Articular; Chondroitin Sulfates; Cross-Sectional Studies; Epitopes; Extracellular Matrix; Humans; Hyaluronic Acid; Knee Injuries; Osteoarthritis, Knee; Young Adult

To find serum markers that may serve as indices for an early diagnosis of degeneration or damage of the articular cartilage.. Twenty-four healthy volunteers, 19 individuals with knee trauma (KT) and 31 with knee osteoarthritis (OA) were evaluated. KT patients were divided into a group (n = 5) with an injury <2 months old (recent KT) and a group (n = 14) with that >2 months old (old KT). Articular cartilage damage was assessed using either arthroscopy or direct observation. Serum concentrations of hyaluronic acid (HA), cartilage proteoglycan aggrecan turnover epitope (CS846) and cartilage oligomeric protein (COMP) were measured using enzyme-linked immunosorbent assay kits and those of keratan sulfate (KS) and chondroitin-6-sulfate (C6S) using high-performance liquid chromatography.. Serum KS in the recent KT group (2095 +/- 594 ng/ml) was significantly higher than that in the old KT group (1373 +/- 418 ng/ml; P = 0.021), and serum COMP in the recent KT group (1572 +/- 182 ng/ml) showed a tendency that was higher than that in the old KT group (1350 +/- 250 ng/ml; P = 0.079). Serum KS in OA patients with Kellgren and Lawrence (KL) grades 0 and I (1456 +/- 334 ng/ml) showed a tendency that was higher than that in OA patients with KL grades II, III and IV (1248 +/- 220 ng/ml; P = 0.084).. The serum concentration of KS correlated with the damage of the articular cartilage and it was significantly increased even at an early stage after the injury.

Topics: Adult; Aged; Arthroscopy; Biomarkers; Cartilage Diseases; Cartilage Oligomeric Matrix Protein; Cartilage, Articular; Chondroitin Sulfates; Extracellular Matrix Proteins; Female; Glycoproteins; Humans; Keratan Sulfate; Knee Injuries; Male; Matrilin Proteins; Middle Aged; Osteoarthritis, Knee; Radiography

Osteoarthritis (OA) influences the levels of free intraarticular glucosaminoglycans (GAG). Little is known about the direction--decrease/increase--of these changes, and information on the correlation between GAG levels and the degree of OA is sparse. Objectives of this study were to investigate the correlation between intraarticular levels of sulphated and unsulphated GAG and the degree of experimental OA, the time course of these changes and whether GAG might be useful as a marker for OA. Twenty-one sheep were randomly assigned to three groups: (1) transsection of the posterolateral bundle of the anterior cruciate ligament, ACL (TD), (2) medial meniscectomy (ME), and (3) meniscectomy and resection of the ACL (MV). During follow-up clinical and radiological examinations were done. After screening for intraarticular effusions, a joint tab was performed and the levels of hyaluronic acid and chondroitin sulphate were measured. The radiological scores differ significantly between group TD and groups ME and MV (P < 0.01). Hyaluronic acid levels in ME and MV are significantly higher than in the controls. Significantly increased levels (P < 0.01) of chondroitin sulphate are found 6 months after ME and 1 year following TD. Clinical consequences: Hyaluronic acid levels--at least in the experimental setting--correspond to a certain degree with osteoarthrotic changes: increasing levels were found along with increasing postoperative interval and increasing grade of OA. Chondroitin sulphate, on the other hand seems, to lend itself as a marker for chondromalacia, in other words for prearthrotic deformities and early stages of OA.

Topics: Animals; Anterior Cruciate Ligament; Anterior Cruciate Ligament Injuries; Biomarkers; Chondroitin Sulfates; Female; Glycosaminoglycans; Hyaluronic Acid; Knee Injuries; Knee Joint; Menisci, Tibial; Osteoarthritis; Postoperative Complications; Sensitivity and Specificity; Sheep; Tibial Meniscus Injuries

To measure the levels of epitope on the chondroitin sulfate (CS) and keratan sulfate (KS) chains of proteoglycan fragments in synovial fluids from injured and contralateral uninjured knees of patients with traumatic cruciate ligament and/or meniscus damage.. Enzyme-linked immunosorbent assays were used to determine the levels of monoclonal antibody epitopes 3-B-3 and 7-D-4 (CS), and 5-D-4 (KS), in paired joint fluids from the injured and uninjured knees of trauma patients.. Levels of the CS epitopes were increased in the trauma joint fluids from most patients, with higher levels of 3-B-3 epitope in 12 of the 16 patients, but the difference did not achieve significance; however, 7-D-4 levels were higher in 15 patients, and the difference was highly significant (P = 0.0005). In contrast, the KS epitope detected by 5-D-4 was decreased in 13 of 15 patients, and the difference was significant (P = 0.0074).. The increased level of 7-D-4 epitope on proteoglycans in joint fluid from injured knees may reflect the response of the articular cartilage to acute trauma resulting in altered expression of specific CS epitopes on cartilage proteoglycans. The fall in KS epitope levels may reflect the synthesis of proteoglycans that have lower KS content.

Topics: Antibodies, Monoclonal; Cartilage, Articular; Chondroitin Sulfates; Enzyme-Linked Immunosorbent Assay; Epitopes; Glycosaminoglycans; Humans; Keratan Sulfate; Knee Injuries; Knee Joint; Synovial Fluid

To investigate the correlation between joint disease and the composition of chondroitin sulfate in the joint fluid, unsaturated disaccharide isomers of chondroitin 4-sulfate (delta di-4S) and chondroitin 6-sulfate (delta di-6S) were measured in joint fluids obtained from patients with osteoarthritis (OA), rheumatoid arthritis (RA), or traumatic arthritis (TA).. These pathologic joint fluids were digested with chondroitinase ABC, and the delta di-4S and delta di-6S produced were determined by high performance liquid chromatography combined with fluorometry.. Total content of delta di-4S plus delta di-6S was 71.8 +/- 30.0 nmoles/ml (mean +/- SD) in OA, 55.4 +/- 29.3 nmoles/ml in RA, and 211 +/- 149 nmoles/ml in TA joint fluids. The ratio of delta di-6S to delta di-4S was 3.81 +/- 0.992 in OA, 1.13 +/- 0.527 in RA, and 5.75 +/- 2.46 in TA joint fluids. Differences between groups were statistically significant.. These results strongly suggest that the levels of chondroitin sulfate isomers and the delta di-6S: delta di-4S ratio in joint fluid reflect the proteoglycan metabolism of joint tissues, particularly of articular cartilage; hence, they could be used to diagnose joint diseases and to predict articular cartilage destruction from such joint diseases.

Topics: Aged; Arthritis; Arthritis, Rheumatoid; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Female; Humans; Hyaluronic Acid; Knee Injuries; Knee Joint; Male; Middle Aged; Osteoarthritis; Synovial Fluid

During studies concerned with the platelet-collagen interaction, it was observed that platelets did not adhere to bovine or human articular cartilage and that cartilage did not induce platelet aggregation in vivo or in vitro. To study the mechanism responsible for this observation, the role of proteoglycans was examined. Purified cartilage collagen proved to be fully active as a platelet aggregant. Addition of small amounts of proteoglycan subunit (PGS) blocked platelet aggregation, whereas chondroitin sulfate, a major glycosaminoglycan component of cartilage matrix, impaired platelet aggregation only at concentrations which resulted in a marked increase in viscosity. Moreover, PGS abolished aggregation of platelets by polylysine but did not prevent aggregation by ADP, suggesting that PGS may block strategically placed lysine sites on the collagen molecule. Treatment of fresh articular cartilage with proteolytic enzymes rendered the tissue active as a platelet aggregant. In vivo experiments demonstrated that surgical scarification of rabbit articular cartilage does not result in adhesion of autologous platelets. Treatment of rabbit knee joints with intraarticular trypsin 1 wk before the injection of blood resulted in adhesion and aggregation of platelets on the surface of the lesions. Since there is evidence from other studies that some degree of cartilage healing may take place after initiation of an inflammatory response, it is postulated that induction of platelet-cartilage interaction may eventuate in cartilage repair.

Topics: Adenosine Diphosphate; Animals; Blood Platelets; Cartilage, Articular; Cattle; Chondroitin Sulfates; Collagen; Humans; Knee Injuries; Papain; Platelet Aggregation; Polylysine; Proteoglycans; Rabbits; Trypsin