Disrupted homeostasis of synovial hyaluronic acid and its associations with synovial mast cell proteases of rheumatoid arthritis patients and collagen-induced arthritis rats

Hyaluronic acid (HA) is the main component of the extracellular matrix (ECM) of joints, and it is important for a lubricating joint during body movement. Degradation is the main metabolic process of HA in vivo. Hyaluronidases (HAase) were known for HA degradation. The inflammation-induced HA rapid-metabolism can reduce HA viscosity and concentration in joints. Mast cells (MC) containing their specific proteases were found in synovium tissue. It is unclear if MC-proteases could be involved in HA degradation pathways. This study aims to explore the correlations between HA concentration vs mast cell proteases, or matrix metalloproteinase-2/9 (MMP-2/9) and to investigate the association of MC-specific proteases with disrupted synovial HA homeostasis in rheumatoid arthritis (RA) or collagen-induced arthritis rats. The synovial fluid samples from no-RA and RA patients were collected; the collagen-induced arthritis (CIA) rat model was established; HA concentration and the activities of MC-protease and MMP-2/9 in the samples were detected, and the correlations were analyzed. In vitro interaction experiment was carried out by mixing MC-proteases with HA to observe the degradation speed. The HA concentrations in synovial fluids were decreased in RA patients and CIA rats compared with those in no-RA subjects or normal rats respectively. The activities of mast cell proteases in synovial fluids were increased and positively correlated with MMP-9, but negatively correlated with HA concentrations. In vitro study, the addition of MC-chymase and tryptase promoted the speed in HA degradation. MC-proteases may influence HA degradation pathway.

     

Biochemical effect of intra-articular injections of high molecular weight hyaluronate in rheumatoid arthritis patients

OBJECTIVE: The purpose of this study was to analyze the biochemical characteristics of synovial fluids after treatment of patients with rheumatoid arthritis (RA) with intra-articularly injected hyaluronate (HA). TREATMENT: In a double-blind study, 13 patients received intra-articular injections of 1% HA in phosphate buffered saline (PBS) while 13 other patients were injected 0.01% HA in PBS, as the control group. Injections were administered once a week for five weeks. METHODS: Clinical efficacy and characteristics of synovial fluid were compared between groups. RESULTS: Significant clinical improvement was observed in the HA-treated group compared to the control group. The viscosity of retained synovial fluid increased. Stringing and HA concentration increased significantly whereas the concentrations of protein and chondroitin sulfate 4 and 6 decreased significantly. The HA molecular weight remained the same in both groups. CONCLUSION: Intra-articular HA injection altered the properties of synovial fluid and proved efficacious for patients with RA.     

Interstitial matrix proteins determine hyaluronan reflection and fluid retention in rabbit joints: effect of protease

Hyaluronan (HA) retention inside the synovial cavity of joints serves diverse protective roles. We tested the hypothesis that HA retention is mediated by the network of extracellular matrix proteins in the synovial lining. Cannulated rabbit knee joints were infused with HA solution with or without pretreatment by chymopapain, a collagen-sparing protease. Trans-synovial fluid escape rate was measured and, after a period of trans-synovial filtration, samples of intra-articular fluid and subsynovial fluid were analysed for HA to assess its trans-synovial ultrafiltration. In control joints, HA ultrafiltration was confirmed by postfiltration increases in intra-articular HA concentration (259 ± 17% of infused concentration) and reduced subsynovial concentration (30 ± 8%; n = 11). The proportion of HA molecules reflected by the synovium was 57–75%. Chymopapain treatment increased the hydraulic permeability of the synovial lining ∼13-fold, almost abolished the trans-synovial difference in HA concentration and reduced the HA reflected fraction to 3–7% ( n = 6; P < 0.001, ANOVA). Structural studies confirmed that chymopapain treatment depleted the matrix of proteoglycans but preserved its collagen. The findings thus demonstrate that HA ultrafiltration and synovial hydraulic permeability are determined by the network of non-collagen, extracellular matrix proteins. This may be important clinically, since protease activity is raised in rheumatoid arthritis, as are HA and fluid escape.     

Different synovial fluid fibronectin levels in rheumatoid variants

Summary A circulating high-molecular-weight glycoprotein called fibronectin plays a part in cell adhesion and migration before phagocytosis and in morphology, differentiation, and metabolism in inflammatory synovial effusions of patients with rheumatic diseases. A technique of nephelometric immunoassay, based on the measurement of an antigen-antibody reaction, was applied to the analysis of fibronectin concentrations in synovial fluids from 20 patients with rheumatoid arthritis (RA) and other diseases (non-RA). RA synovial fluids have a significantly higher concentration than the specimens obtained from Yersinia arthritis patients (n=12). The mean concentration of other synovial fluids, from 12 patients with osteoarthritis of the knees, did not significantly differ from the synovial fluids of control values obtained from patients who underwent meniscectomy. There was a considerably negative correlation between fibronectin levels and overall indices of inflammatory activity, such as Ritchie articular indices or a whole number of painful rheumatoid arthritis joints. However, a particularly distinct correlation was obtained when raised fibronectin levels were compared with the inflammatory activity of the knee joint, from which the specimen was aspirated. Thus, these findings suggest that the measurements of fibronectin in synovial fluid may be of some differential-diagnostic value in rheumatoid variants, but may only serve as an indicator of inflammatory activity if the joint, from which the specimen is obtained, is taken into account.Dedicated to Prof. Dr. N. Zöllner on his 60th birthdaySupported in part by the Deutsche Forschungsgemeinschaft, Go 236/8–9     

Pharmacokinetics of tiaprofenic acid in normal rabbits and rabbits subjected to joint immobilisation

In a previous study, tiaprofenic acid (TA) was administered daily over a 30-day period at 5 and 10 mg/kg of body weight subcutaneously (s.c.) to animals with arthritis induced by immobilisation. The 10 mg/kg dose exacerbated the loss of proteoglycan from joint cartilage but the 5 mg/kg dose showed protective effects on articular cartilage. These results led us to investigate the concentration of TA achieved in synovial fluid of both the immobilised and non-immobilised rabbit joints after single s.c doses of 5 or 10 mg/kg. The halflives of elimination of TA from the synovial fluids of the immobilised joints were 1.27 and 1.07 h after the 5 and 10 mg doses, respectively, and 0.66 and 0.39 h in the non-immobilised contralateral joints. Clearances from synovial fluid to plasma were found to be 0.41 and 0.55 ml/h/kg from the immobilised joints after the 5 and 10 mg doses, respectively, and 0.11 and 0.25 ml/h/kg from the non-immobilised contralateral joints. The peak concentration of TA achieved in synovial fluid of immobilised knee joints after a single s.c. injection of 10 mg/kg was approximately two times greater than the concentrations achieved after administration of 5 mg/kg by the same route and two to six times greater than the levels achieved after oral administration of TA at 600 mg/day in humans.     

Concentration polarization of hyaluronan on the surface of the synovial lining of infused joints

Hyaluronan (HA) in joints conserves the lubricating synovial fluid by making trans-synovial fluid escape almost insensitive to pressure elevation (e.g. effusions, joint flexion). This phenomenon, 'outflow buffering', was discovered during HA infusion into the rabbit knee joint cavity. It was also found that HA is partially reflected by the joint lining (molecular sieving), and that the reflected fraction R decreases as trans-synovial filtration rate Q is increased. It was postulated therefore that outflow buffering is mediated by HA reflection. Reflection creates a HA concentration polarization layer, the osmotic pressure of which opposes fluid loss. A steady-state, cross-flow ultrafiltration model was previously used to explain the outflow buffering and negative R-vs.-Q relation. However, the steady-state, cross-perfusion assumptions restricted the model's applicability for an infused, dead-end cavity or a non-infused joint during cyclical motion. We therefore developed a new, non-steady-state model which describes the time course of dead-end, partial HA ultrafiltration. The model describes the progressive build-up of a HA concentration polarization layer at the synovial surface over time. Using experimental parameter values, the model successfully accounts for the observed negative R-vs.-Q relation and shows that the HA reflected fraction ( R ) also depends on HA diffusivity, membrane area expansion and the synovial HA reflection coefficient. The non-steady-state model thus explains existing experimental work, and it is a key stage in understanding synovial fluid turnover in intact, moving, human joints or osteoarthritic joints treated by HA injections.     

Characteristics of polyarthritis in rabbits by hyperimmunization with attenuated Enterococcus faecalis.

To produce polyarthritis and rheumatoid factor like substance (RFLS), rabbits were hyperimmunized intravenously with 0.02% thimelosal (TMS)-treated Enterococcus faecalis (E. faecalis) as a persistent bacterial flora. Swelling of knee joints occurred at a rate of 41% (27/66), and of shoulder joints at a rate of 25% (17/66) while that of elbow joints occurred at a rate of 4.5% (3/66). On culturing of knee joint fluids, no colonies appeared while 2/4 fluid specimens from the shoulder joints gave positive colonies for 78 days after the first immunization; thereafter, no colonies appeared. On histological examination, in early stages, acute inflammatory reactions with degenerative changes of synovial tissue was observed. In later stages, chronic inflammatory changes, proliferation of synovial cells with pannus formation, destruction of articular cartilage and subchondral bone were observed. RFLS titer showed bi-phasic peaks at 11 days and 41 days after the first immunization. A high incidence of polyarthritis, particularly knee joints, occurred. Thus, hyperimmunization with attenuated E. faecalis as a normal intestinal flora may provide an animal model of chronic polyarthritis.     

Modulation of Hyaluronan Fragmentation by Interleukin-1 Beta in Synovial Membrane Cells

Hyaluronan (HA) plays a crucial role in the lubricating and buffering properties of synovial fluid. The purpose of this study was to examine the effects of interleukin (IL)-1β on HA degradation in cultured synovial membrane cells. The rabbit synovial membrane cell line HIG-82 was cultured with and without IL-1β. The amounts of HA of varying molecular weights in the medium were analyzed using high-performance liquid chromatography, the mRNA levels of HA synthase (HAS) and hyaluronidase (HYAL) were analyzed by means of real-time PCR, and HYAL activity was analyzed by HA zymography. The amounts of HA with a molecular weight lower than 300 kDa, and between 300 and 1900 kDa, in the culture medium of HIG-82 cells were significantly higher in the presence of IL-1β. However, the amount of HA with a molecular weight greater than 1900 kDa was significantly lower in the presence of IL-1β. Both HAS2 and HAS3 mRNA levels were upregulated by treatment with IL-1β. So, too, were the levels of HYAL1 and HYAL2 mRNA, which resulted in enhanced HYAL activity. However, HYAL activity was inhibited by transfection of HYAL2-siRNA. Our results suggest that IL-1β is a crucial factor in the fragmentation of HA in inflammatory joints.     

Hyaluronic Acid for the Treatment of Osteoarthritis in all Joints Except the Knee

Background

The use of intra-articular hyaluronic acid (HA) is a well known treatment in patients with knee osteoarthritis (OA). In other joints, less evidence is available about the efficacy of treatment with intra-articular HA. HA is also used intra-articularly in the metatarsophalangeal-1 joint, the ankle, the hip, the sacroiliac joint, the facet joints, the carpometacarpal-1 joint, the shoulder and the temporo-mandibular joint. In this systematic review we include all prospective studies about the effects of intra-articular HA in the above-mentioned joints. Its use in the knee joint, however, will be discussed in a separate article in this journal.

Methods

A systematic review was conducted using databases including MEDLINE, Cochrane Database of Systematic Reviews, Cochrane Clinical Trial Register, and EMBASE.

Results

After performing a solid systematic review using a rigid methodology and trying to pool the outcomes of different studies, we noticed that, compared with baseline, there is statistical evidence for a positive effect of intra-articular HA. However, there is limited evidence HA is superior to placebo and no evidence that intra-articular HA is better than corticosteroids or other conservative therapies.

Conclusion

Our recommendation for future research is that one should focus on adequately powered randomized trials comparing HA treatment with other types of intra-articular or conservative treatment. We think it is useless to further perform and publish (large) non-comparative prospective studies about the use of HA in the treatment of problems caused by OA. It is well perceived that HA exerts positive effects in the treatment of OA, but up to now there is no (strong) evidence available that HA is superior to other treatments of OA such as corticosteroids, physiotherapy or other conservative measures.     

滑膜软骨瘤病的X线病理分析

目的 探讨滑膜软骨瘤病的临床、病理与X线联系及诊断价值．方法 回顾性分析了17例滑膜软骨瘤病的临床、病理及X线表现．结果 本病好发于下肢大关节，多为单关节．膝关节10例(双侧2例)、肩关节3例、髋关节、肘关节、腕关节、指掌关节各1例．主要临床表现：关节肿痛、弹响(6例)，绞锁(3例)，触及可活动或不活动性硬块(9例)．X线特点：在关节内及其周围见数目、大小、密度、形态不一的钙化或骨化游离体．结论 正确判断和认识关节内游离体来源与发展的病理变化及X线表现对诊断本病有较重要的价值．     

Postural coordination patterns associated with the swinging frequency of arms

Voluntary arm movements frequently perturb body equilibrium in an upright posture. The motions of leg joints need to be coordinated according to the properties of voluntary arm movements in order to maintain body equilibrium, and this may cause a change in postural pattern. The purpose of this study was to determine whether the kinematic pattern generation of upright posture is influenced by a change in the swinging frequency of arm movements and whether the pattern generation is correlated with a change in joint torque about the shoulder joint. Four male subjects in an upright posture were instructed to swing their arms at seven different frequencies, determined by the maximum swinging frequency of each subject (35%max, 40~60%max, 65%max). Segment rotations around the shoulder, hip, and ankle joints were analyzed at kinematic and kinetic levels. The results of kinematic analysis indicated that tight coupling between motions of the shoulder and hip joints was generated in lower-frequency trials (under 40–45%max), whereas tight coupling between motions of the shoulder and ankle joints was generated in higher frequency trials (more than 40–45%max). Furthermore, the results of kinetic analysis revealed that changes in the joint torque patterns about the shoulder and hip joints occurred in trials at 40–45%max. The mean value of 40–45%max was close to the eigenfrequency of each subject's arm. We concluded that (1) postural patterns associated with a gradual change in the swinging frequency of the arms can be divided into two coordination modes (a hip-shoulder in-phase mode and an ankle-shoulder in-phase mode), and (2) these two patterns may be divided by the eigenfrequency of the arm. Electronic Publication     

The painful diabetic shoulder

Different types of shoulder affection were studied in 62 diabetic patients with shoulder pain. Three groups of shoulder joint disorder were found: painful shoulder with restricted mobility (62%), tendinitis without mobility restriction (27%), and a small group with mixed diagnoses. Sixty per cent had hand symptoms and 38% had restricted mobility of their hip joints. High frequencies of retinopathy and neuropathy were found. Affection of the shoulder joint was seen with almost the same frequency in insulin-dependent as in non-insulin-dependent patients, but after a shorter duration of diabetes in the latter. A group of patients with the triad shoulder pain, hand symptoms and restricted mobility of the hip joints had a significantly higher frequency of proliferative retinopathy than patients with shoulder pain only. The long duration of diabetes, the high frequency of insulin treatment and classical late complications indicate that diabetic patients with painful shoulder and restricted mobility are suffering from clinically advanced diabetes mellitus.     

A large family with multiple epiphyseal dysplasia linked to COL9A2 gene

We describe a large family, including 54 affected individuals, with multiple epiphyseal dysplasia (MED) with involvement of the peripheral joints only. In this family, a mutation in the COL9A2 gene was detected. Every affected person has involvement of the knee joints. Other involved joints are the elbow, ankle, wrist, hand, and feet joints. Involvement of the shoulder or hip joints is never seen after adolescence. The height of the affected individuals is short to normal; spinal involvement is never seen. The penetrance of the gene is complete. However, the expression of the gene is highly variable. Am. J. Med. Genet. 77:234–240, 1998. © 1998 Wiley-Liss, Inc.     

Soluble Forms of P-Selectin and Intercellular Adhesion Molecule-3 in Synovial Fluids

A number of adhesion molecules have been identified in synovial tissues of patients with rheumatoid arthritis (RA). Some of them are upregulated and may play an important role in the inflammatory processes of the diseased joint. In addition to synovial tissue cell surface expression, synovial fluids contain soluble forms of many adhesion molecules, such as intercellular adhesion molecule-1, E-selectin (sE-selectin), and L-selectin. In this study, we investigated the expression of soluble P-selectin (sP-selectin) and intercellular adhesion molecule-3 (sICAM-3) in synovial fluids from patients with RA, osteoarthritis (OA), and other forms of arthritis. sP-selectin and sICAM-3 levels in RA synovial fluids were significantly increased compared to those in OA. The levels of sP-selectin in synovial fluids correlated with sICAM-3 and sE-selectin in synovial fluids. The levels of sICAM-3 in synovial fluids correlated with synovial fluid leukocyte counts and erythrocyte sedimentation rate.In vitro,synovial fluid mononuclear cells produced sICAM-3 spontaneously. Elevated levels of sP-selectin and sICAM-3 in RA synovial fluids compared to OA may indicate inflammatory interactions between endothelial cells, leukocytes, and other synovial cells in the diseased joint.     

Oligoclonal immunoglobulins and smooth muscle antibodies in arthritic joints.

In twelve synovial fluid/serum pairs from patients with various types of seronegative polyarthritis, homogeneous gamma-bands by agarose gel electrophoresis were found in seven of the synovial fluids and in only one of the sera. In six of the fluids with gamma-bands, smooth muscle antibodies (SMA) were also present, usually in a titre identical to that in serum. In fluids with no gamma-bands, no SMA were detected. In forty synovial fluid/serum pairs from paitients with seropositive rheumatoid arthritis, no gamma-bands were detected in the synovial fluids, and SMA were present in only three pairs. Absorption and inhibition experiments did not give evidence that the SMA activity in seronegative polyarthritis was confined to the gamma-bands in the synovial fluids. The SMA activity in the fluids seemed to be directed against both actin and 'non-actin' muscular antigens. The association between locally produced oligoclonal immunoglobulins and possible locally produced SMA with differnet electrophoretic mobility suggests that in some of thes patients there is a local synovial production of oligoclonal antibodies with different specificities. Thus, even if the results may indicate a local virus infection in some arthritic joints, they may also be dur to an unspecific local stimulation of B cells or to a specific antigen stimulation combined with an unspecific co-activation of other antibody-producing cells.     

Applications and emerging trends of hyaluronic acid in tissue engineering,as a dermal filler and in osteoarthritis treatment

Hyaluronic acid (HA) is a naturally occurring biodegradable polymer with a variety of applications in medicine, including scaffolding for tissue engineering, dermatological fillers and viscosupplementation for osteoarthritis treatment. HA is available in most connective tissues in body fluids such as synovial fluid and the vitreous humor of the eye. HA is responsible for several structural properties of tissues as a component of extracellular matrix and is involved in cellular signaling. Degradation of HA is a stepwise process that can occur via enzymatic or non-enzymatic reactions. A reduction in HA mass or molecular weight via degradation or slowing of synthesis affects physical and chemical properties such as tissue volume, viscosity and elasticity. This review addresses the distribution, turnover and tissue-specific properties of HA. This information is used as the context for considering recent products and strategies for modifying the viscoelastic properties of HA in tissue engineering, as a dermal filler and in osteoarthritis treatment.     

Boiogito,a Kampo medicine,improves hydrarthrosis in a rat model of knee osteoarthritis

Background

Hydrarthrosis, which is associated with knee pain and limited range of motion, decreases the quality of life (QOL) of patients with osteoarthritis (OA). The Kampo medicine boiogito is prescribed for the treatment of knee OA with hydrarthrosis; however, its precise mechanisms of action remain unknown. The purposes of this study were to assess the pharmacological effects of boiogito and its mechanisms of action on joint effusion in rats with surgically induced OA.

Methods

A rat OA model was produced by transecting the anterior (cranial) cruciate ligament, medial collateral ligament, and medial meniscus in the right knee joints of 7-week-old female Wistar rats. The rats were given chow containing boiogito (1 or 2 %) or indomethacin (0.002 %) for 4 weeks after surgical transection. Levels of interleukin-1β (IL-1β) and hyaluronic acid (HA) were measured by enzyme-linked immunosorbent assay. Knee joint pain was assessed using an incapacitance tester. Osmotic water permeability in cultured rabbit synovial cells was assessed using stopped-flow analysis.

Results

Increased synovial fluid volume and knee joint pain were observed in rats with surgically induced OA. In rats with OA, levels of IL-1β and HA in the articular cavity were higher but concentration of HA in synovial fluid was lower than in sham-operated rats, suggesting excessive synovial fluid secretion. Administration of boiogito improved hydrarthrosis, IL-1β, and HA concentrations and alleviated knee joint pain in rats with OA. Indomethacin reduced IL-1β and knee joint pain but failed to improve hydrarthrosis or HA concentration in rats with OA. Osmotic water permeability in synovial cells, which is related to the function of the water channel aquaporin, was decreased by treatment with boiogito.

Conclusion

Boiogito ameliorates the increased knee joint effusion in rats with OA by suppressing pro-inflammatory cytokine IL-1β production in the articular cavity and regulating function of water transport in the synovium. The improvement of hydrarthrosis by boiogito results in the increased HA concentration in synovial fluid, thus reducing joint pain. Boiogito may be a clinically useful treatment of QOL in patients with OA with hydrarthrosis.

     

Thin film rheology and lubricity of hyaluronic acid solutions at a normal physiological concentration

Using a surface forces apparatus to measure forces, and optical (multiple beam) interferometry to measure surface shapes and separations (to +/-1 A), the normal, viscous, and shear (lubrication) forces between smooth mica surfaces in aqueous hyaluronic acid (HA) solutions were measured. The experimental conditions of loading pressures, pH, and HA concentration were set to closely correspond to physiological human knee-joint conditions. From the force and optical (refractive index) measurements, it was concluded that, like other negatively charged polyelectrolytes, HA does not naturally adsorb on the mica surface which is hydrophilic and weakly negatively charged at physiological conditions: the polymer solution exhibits the bulk viscosity (22.5 +/- 1.5 cP) for films thicker than about 0.4 miccrom of the polymer, whereas for thinner films, the viscosity decreases monotonically toward the value of the pure electrolyte solution (1 cP) as HA is extruded from between the surfaces. This is indicative of a repulsive "depletion" interaction of HA with each mica surface and to a weakly attractive polymer-mediated force between the two surfaces. Thus, free HA in synovial fluid is not expected to act as a good "boundary lubricant." Relaxation measurements on approaching and receding surfaces in HA solutions were also performed, and it is shown that the presence of HA in the bulk solution can improve "hydrodynamic" modes of lubrication, for example, by assuaging the compression stroke. The study includes information that is beneficial to researchers working with biomaterials viscosupplementation devices.     

等动肌力矩的增龄性发育特征

背景：少年时期肌肉力量是否能够得到平衡发育对其生长发育过程中起重要作用。 目的：探索少年的肌力发展的生物力学规律。 方法：利用Kinitech等速测力系统对中小学男女生各30名双侧肩、髋关节屈伸肌群进行等动肌力测试,测试速度为60 (°)/s,测试次数为屈伸6次。 结果与结论：所有测试者的髋、肩关节屈伸肌的相对峰力矩随测试速度的增大而逐渐减小;且随年龄增长肩、髋关节屈伸肌的相对峰力矩均同步增大;小学生在不同测试速度状态下男生髋、肩关节相对峰力矩均大于女生(P < 0.05),但中学生髋、肩关节相对峰力矩没有性别差异。     

Immune complexes in early arthritis. II. Immune complex constituents are synthesized in the synovium before rheumatoid factors.

Synovial fluids and paired sera taken from patients either before, after or at the time of diagnosis of definite rheumatoid arthritis (RA) were compared with samples from patients with unclassified inflammatory arthropathies (IA). Raised levels of immune complexes (IC) were detected in some RA patients by C1q binding activity but in the majority of both RA and IA patients by the platelet aggregation test; levels were usually higher in joint fluids than in sera. IgM rheumatoid factors (RF) and IgA RFs were lower in synovial fluids but IgF RF levels were similar in matched samples. Synovial fluid to serum albumin ratios were used to estimate synovial permeability (inflammation) and then to calculate which patients synthesized macromolecules locally in the synovium. Local synthesis of RFs was detected in a greater proportion of RA than IA patients and only two patients formed RFs locally in the first months of symptoms. Half the patients in both groups however appeared to synthesize or trap IC constituents and in many patients there was evidence of local synthesis within 6 months after their symptoms had started. We conclude that local synthesis of large amounts of RFs is uncommon in the early stages of RA but that IC of unknown composition are synthesized or localized in the affected joints of many patients with RA and inflammatory arthropathies shortly after their symptoms appear.