Knee Synovial Membrane Research Articles

SARS-CoV-2 in the knee joint: a cadaver study.

Despite the considerable research efforts being made to learn more about COVID-19, little is known about the presence of SARS-CoV-2 genetic material in biological fluids other than respiratory droplets, blood, and feces. The aim of this post-mortem study was to assess the presence of SARS-CoV-2 RNA in the knee synovial fluid, synovial tissue, and bone tissue of COVID-19 patients in order to discover whether the joint is a possible route of transmission during orthopaedic surgical procedures, and clarify the possible role of SARS-CoV-2 as a directly arthritogenic virus. Post-mortem synovial fluid, synovial tissue and bone tissue samples were collected from the knees of five patients who died of COVID-19 in our hospital between September and October 2020, and analysed for the presence of SARS-CoV-2 using a commercial real-time polymerase chain reaction (RT-PCR) panel. Quantitative RT-PCR was used to test post-mortem nasopharyngeal swabs of all of the patients. No SARS-CoV-2 RNA was detected in any of the knee samples, despite the positivity of the throat swab. Our findings indicate that SARS-CoV-2 was not detected in knee synovial fluid, synovial membrane or bone. This makes it unlikely that these are potential sources of contagion, and suggests that SARS-CoV-2 is not directly arthritogenic.

AB0671 NO EVIDENCE OF SARS-COV-2 IN THE KNEE JOINT: A CADAVER STUDY

Background:Despite the considerable research efforts being made to learn more about COVID-19, little is known about the presence of SARS-CoV-2 genetic material in biological fluids other than respiratory droplets, blood, and feces 1,2. In particular, little is known about the presence of SARS-CoV-2 in the joints either post mortem3 or in vivo4. To the best of our knowledge, only Lopéz-Gonzalez et al.5 have published a description of acute arthritides occurring during hospitalisation due to COVID-19, and they did not find any SARS-CoV-2 genetic material in the patients’ synovial fluid samples.Objectives:The aim of this post mortem study was to assess the presence of SARS-CoV-2 RNA in the knee synovial fluid, synovial tissue, and bone tissue of COVID-19 patients in order to discover whether the joint is a possible route of transmission during orthopaedic surgical procedures, and clarify the possible role of SARS-CoV-2 as a directly arthritogenic virus.Methods:Post mortem synovial fluid, synovial tissue and bone tissue samples were collected from the knees of five patients who died of COVID-19 in our hospital between September and October 2020, and analysed for the presence of SARS-CoV-2 using a commercial real-time polymerase chain reaction (RT-PCR) panel. Quantitative RT-PCR was used to test post mortem nasopharyngeal swabs of all of the patients.Results:No SARS-CoV-2 RNA was detected in any of the knee samples, despite the positivity of the throat swab.Conclusion:Our findings indicate that SARS-CoV-2 was not detected in knee synovial fluid, synovial membrane or bone. Therefore, our results suggest that all healthcare professionals performing surgical procedures on the joints of COVID-19 patients are exposed to a risk of contagion due to exposure to respiratory droplets, blood and body fluids, but not to direct exposure to joint- or bone-related tissues. Furthermore, given that some case reports of arthritis in COVID-19 patients 5-8 show that it is possible that COVID-19 patients display viral-mediated arthralgias and arthritis, the absence of the virus in the knee highlighted by our study suggests that it is unlikely that SARS-CoV-2 has a direct inflammatory action on the joint, but it could induce an inflammation-related reaction, manifesting as a reactive arthritis 9.

In vivo protective effect of adipsin-deficiency on spontaneous knee osteoarthritis in aging mice.

The adipokine adipsin is an emerging mediator of human osteoarthritis (OA) progression. Here, we investigated its in vivo role in the development of spontaneous OA in aging mice. We compared articular knee joint morphology, histology in knee cartilage, synovial membrane, subchondral bone, meniscus, and anterior cruciate ligament (ACL); and chondrogenesis in the ACL from adipsin-deficient (Df-/-) and wild-type (Df+/+) 20-week- and 20-month-old mice. Serum levels of a panel of adipokines, inflammatory factors, and metalloproteases known to be implicated in OA were investigated. Data first revealed that the early manifestation of OA appeared in the ACL of 20-week-old mice, progressing to severe alterations in the 20 month-old wild-type mice. Further results demonstrated that adipsin-deficiency protected the articular tissues from spontaneous OA progression and triggered significantly higher serum levels of the adipokines adiponectin and FGF-21 while lowering levels of the inflammatory factor interleukin 6 (IL-6) in both young and old mice. This work further underlines the clinical relevance of adipsin as a novel therapeutic approach of human OA. Moreover, this study shows the potential beneficial effect of the adipokine FGF-21 against OA, and provides support for this factor to be a new biomarker and/or target of primary OA therapeutic avenues.

Intra articular methylprednisolone acetate injection efficacy in the treatment of primary knee osteoarthritis at Hue Central Hospital and Hospital of Hue University of Medicine and Pharmacy

Introduction/Background: Osteoarthritis is still an incurable disease, the current treatment is to reduce clinical symptoms. The intra articular glucocorticoid injection in knee osteoarthritis makes the inflammatory process self-limiting and improves pain symptom. Objectives: To evaluate the efficacy and safety of intra articular methylprednisolone acetate injection in treatment of primary knee osteoarthritis. Materials and Methods: Prospective descriptive study of 88 knee joints (34 joints in study group, 54 joints in control group) was diagnosed with primary osteoarthritis according to the American College of Rheumatology (ACR) classification criteria and in Kellgen & Lawrence grade II, III. The patient was assessed about clinical features, subclinical features and the VAS score, Lequesne index at the initial of the study and 7 days later. Results: The analgesic effect was higher in the group of intra articular methylprednisolon injection with statistically significant difference (∆VAS of the study group and the control group were 38.65 ± 9.25; 25.11 ± 9.26, respectively, with p < 0.05). The improving of knee function was higher in the group of intra articular methylprednisolon injection with statistically significant difference (∆Lequesne of the study group and the control group were 7.00 (5.00 – 8.00); 4.00 (3.00 – 5.00), respectively, with p < 0.05). The incidence of complications in the study group was 1/34 (2.94%) with post – injection pain. The improvement of VAS is better in patient with severe pain than those with moderate pain (p < 0.05) and correlated to the initial value of Lequesne (r2 = 0,162, respectively, with p < 0.05). The improvement of Lequesne correlated to the joint fluid thickness in ultrasound (r2 = 0.102, p < 0.05). There is no relation between efficacy of treatment and site of knee joint, radiology stage and synovial membrane thickness (p > 0.05). Conclusions: The intra articular methylprednisolone acetate injection in treatment of primary knee osteoarthritis is a safe treatment and has significant, clearly effect in pain relief and physical function improving after 7 days. The improvement of VAS is better in the patient with higher initial value of VAS, Lequesne and the improvement of Lequesne is better in the patients with more synovial thickness in ultrasound. Key words: Knee osteoarthritis, intraarticular injection, methylprednisolon acetate

Effects of ID-CBT5101 in Preventing and Alleviating Osteoarthritis Symptoms in a Monosodium Iodoacetate-Induced Rat Model.

Osteoarthritis is a disease that affects the articular cartilage and osseous tissue, and can be worsened by aging, overweight status, and post-traumatic arthritis. The present study aimed to evaluate the effect of ID-CBT5101 (tyndallized Clostridium butyricum) on bone metabolism and the inflammatory response in a monosodium iodoacetate-induced rat model of osteoarthritis. ID-CBT5101 was administered orally at doses of 108 or 1010 CFU/day for 2 weeks before direct injection of monosodium iodoacetate (3 mg/50 μl of 0.9% saline) into the intra-articular space of the rats' right knees. The rats subsequently received the same doses of oral ID-CBT5101 for another 4 weeks. We evaluated the treatment effects based on serum biomarkers, mRNA expression, morphological and histopathological analyses of the knee joints, and weight-bearing distribution analysis. Compared with those in control rats, the ID-CBT5101 treatments significantly reduced the serum concentration of inflammation and bone metabolism markers (i.e., COX-2, IL-6, LTB4, and COMP), and significantly increased the concentration of IFN-γ and glycosaminoglycans. In addition, the ID-CBT5101 treatments inhibited the mRNA expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases (i.e., MMP-2, MMP-3, MMP-9, MMP-13, TIMP-1, and TIMP-2). Furthermore, the ID-CBT5101 treatments effectively preserved the knee cartilage and synovial membrane, and significantly decreased the amount of fibrous tissue. Moreover, compared with that of the negative control group, the ID-CBT5101 treatments increased the weight-bearing distribution by ≥20%. The results indicate that ID-CBT5101 prevented and alleviated osteoarthritis symptoms. Thus, ID-CBT5101 may be a novel therapeutic option for the management of osteoarthritis.

The infrapatellar fat pad and the synovial membrane: an anatomo-functional unit.

The infrapatellar pad, a fibro-adipose tissue with peculiar microscopic and mechanical features, is gaining wide attention in the field of rheumatological research. The purpose of this descriptive review is to summarize the most recent published evidence on the anatomic, physiologic and biomechanical inter-relationship between the infrapatellar fat pad and the knee synovial membrane. As an extrasynovial tissue, the infrapatellar fat pad does not directly interact with the articular cartilage; based on its location in close contact with the synovial membrane, and due to the metabolic properties of adipose tissue, it may influence the behavior of the synovial membrane. In fact, considering evidence of macroscopic and microscopic anatomy, the infrapatellar fat pad is the site of insertion of the infrapatellar and medial synovial plicae. Also biochemically, there is much evidence highlighting the interaction among these two structures; in the case of inflammation, the mutual interplay is ascribable to the release of pro-inflammatory mediators stimulating the proliferation of inflammatory cells and promoting tissue modifications in both. All these assumptions could support the emerging idea that the infrapatellar fat pad and the synovial membrane may be considered a morpho-functional unit.

KNEE CARTILAGE AND SYNOVIAL MEMBRANE STRUCTURAL CHANGES DURING TIBIA DISTRACTION WITH PLATING

Purpose of the study — to analyze the changes in knee articular cartilage and synovial membrane during distraction external fixation of the tibia in combination with plating.Material and methods. Articular cartilage and synovial membrane of the knee joint were studied using histomorphometry methods in 9 mongrel dogs during distraction external fixation of the tibia combined with plating. Tibia and fibula osteotomies were performed at the border of middle and upper third, plate was fixed on tibia diaphysis. Lengthening was achieved at rate of 1 mm per day in four stages during 21–28 days. Animals were withdrawn from experiment in 30 and 90 days. After autopsy of knee joints the authors excised sections of synovial membrane from suprapatellar area, articular cartilage with underlying subchondral bone from loadable surface of femoral condyles. Thickness of articular cartilage, its area and volumetric density of chondrocytes was measured, proportion of chondrocytes within isogenic groups from the overall number of chondrocytes as well as proportion of empty lacunae. In synovial membrane the authors measured thickness of surface layer and numeric density of micro vessels. Articular cartilage of 5 intact animals was used as a control group.Results. After 30 days of plate fixation a hyperplasia of the integument layer, mild synovitis, and hypervascularization were observed in synovial membrane. Density of micro vessels increased to 363.93±33.71 (control group — 335.05±28.88). The authors also observed subperineural and endoneural edema as well as destruction of nerve fibers in subsynovial layer. Articular cartilage retained the zonal structure. Destructive changes were manifested by fibers separation in the superficial part of surface zone and by partial loss of chondrocytes. The following parameters were reduced: cartilage thickness, area and volumetric density of chondrocytes, proportion of isogenic groups; empty lacunae exceeded the values in controls by 18.2%. After 90 days of plate fixation, thinning of the cover layer of synovial membrane was reported. The numerical density of micro vessels decreased to 325.81±36.39. In nerves of subsynovial layer the edema and vacuolization of myelin sheaths of preserved nerve fibers as well as the formation of Büngner bands in place of degenerating ones were detected. Synovitis was not observed. Fibers separation of extracellular substance in upper superficial part of the surface cartilage zone was retained. There was a tendency to increase in cartilage thickness, area and volumetric density of chondrocytes, proportion of isogenic groups; and to decrease of empty lacunae number.Conclusion. The histological changes in the articular cartilage during distraction external fixation of the tibia in combination with plating corresponded to the initial stages of osteoarthritis of grades 1—2 according to histological classification of International Society for Study of Osteoarthritis (2006) and were accompanied by hypovascularization and denervation of synovial membrane.

Immunohistochemical characterization of early and advanced knee osteoarthritis by NF-κB and iNOS expression.

This study was performed to determine the differences in grade of synovitis and expression of NF-κB and iNOS in knee synovial membrane between early and advanced stage of osteoarthritis (OA). Thirty synovial membrane intra-operative biopsies of patients (ten controls, ten with early and ten with advanced OA according to Kellgren-Lawrence radiological score) were immunohistochemically (NF-κB and iNOS) and hystologically (Krenn synovitis score) analyzed and correlated to WOMAC clinical score and pain duration. Krenn synovitis score of patients with radiologically early OA was significantly higher than in patients with advanced OA (p < 0.001). NF-κB expression in both synovial intima (p < 0.001) and subintima (p < 0.001) was also higher in early OA. iNOS expression in subintima was significantly higher in early than in advanced OA (p < 0.001), while in intima iNOS showed no statistical difference between groups (p = 0.07). The lymphocytic nodules, located in synovial subintima, were significantly higher in advanced OA when compared to early OA (p = 0.006) and the control group (p < 0.001). These results suggest that in early OA, there is a localized inflammation of the synovial membrane with high expression of NF-κB and iNOS. In advanced OA, number of expressed factors is reduced, with the exception of intima cells that highly express iNOS, reflecting the ongoing localized inflammatory process of lower degree. In advanced OA, the density of the resident cells is reduced and lymphocytic nodules appear, confirming the important role of adaptive immunity in later OA stage. Clinical significance of this study is better understanding possibilities of preventive measures for synovitis and OA advancement. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1990-1997, 2017.

Iron overload in a murine model of hereditary hemochromatosis is associated with accelerated progression of osteoarthritis under mechanical stress

Hereditary hemochromatosis (HH) is a disease caused by mutations in the Hfe gene characterised by systemic iron overload and associated with an increased prevalence of osteoarthritis (OA) but the role of iron overload in the development of OA is still undefined. To further understand the molecular mechanisms involved we have used a murine model of HH and studied the progression of experimental OA under mechanical stress. OA was surgically induced in the knee joints of 10-week-old C57BL6 (wild-type) mice and Hfe-KO mice. OA progression was assessed using histology, micro CT, gene expression and immunohistochemistry at 8 weeks after surgery. Hfe-KO mice showed a systemic iron overload and an increased iron accumulation in the knee synovial membrane following surgery. The histological OA score was significantly higher in the Hfe-KO mice at 8 weeks after surgery. Micro CT study of the proximal tibia revealed increased subchondral bone volume and increased trabecular thickness. Gene expression and immunohistochemical analysis showed a significant increase in the expression of matrix metallopeptidase 3 (MMP-3) in the joints of Hfe-KO mice compared with control mice at 8 weeks after surgery. HH was associated with an accelerated development of OA in mice. Our findings suggest that synovial iron overload has a definite role in the progression of HH-related OA.

The discovery of the synovial lymphatic stomata and lymphatic reabsorption in knee effusion.

To illustrate the mechanism of lymphatic reabsorption in knee joint effusion. The current investigation employed transmission electron microscopy (TEM) and scanning electron microscopy (SEM) techniques to reveal the ultrastructure of the knee synovial membrane in New Zealand rabbits and human. Ultrastructural changes of the synovial lymphatic stomata were observed by using trypan blue absorption and sodium hydroxide (NaOH) digestion methods, and the animal models of synovitis. New Zealand rabbits and human synovial membranes were composed of two types of synovial cells: type A and type B. No lymphatic stomata were found among type A synovial cells, whereas lymphatic stomata with the diameters ranging 0.74-3.26 µm were found in type B synovial cells, and some stomata were closed. After the NaOH digestion, a number of sieve pores, similar to lymphatic stomata in size and shape, were observed in the dense fibrous connective tissue underneath the type B synovial cells. After injecting trypan blue into the rabbit knee joint cavity, absorption of trypan blue through the lymphatic stomata was observed, suggesting the absorption function of the synovial lymphatic stomata. In the rabbit knee joint synovitis models, the synovial lymphatic stomata diameter enlarged. Some macrophages migrated from the lymphatic stomata, indicating that the synovial lymphatic stomata were involved in the joint effusion absorption and inflammatory response. Our study is the first to report the existence of synovial lymphatic stomata in the New Zealand rabbits and human knee joints. Lymphatic stomata may have an important role in the reabsorption of joint effusion.

Structural reorganization of the knee joint cartilage and synovium during diaphyseal femoral fracture management using intramedullary locked nails ^perime^al and morphological study)

The purpose - to study structural changes in knee articular cartilage and synovial membrane during locking intramedullary osteosynthesis of femoral fractures. Material and methods. The study tested structural changes in the articular cartilage of the femoral condyles and synovium during femoral fracture repair under the conditions of intramedullary locked nailing. Transverse diaphyseal femoral fractures were modeled in 12 adult mongrel dogs. Synovitis severity was assessed with a V. Krenn scale. Wilcoxon Test was used for testing hypothesis. Results. In the first group (n = 6) fractures were fixed with locked intramedullary nails ILoc (Biomedtrix, USA), while the same type of fixation in group 2 (n = 6) was performed on 4 th day postoperatively after reduction by skeletal traction. It was revealed that in the first group a unified medullary cavity was seen by day 70 and the cortical layer at the fracture site approximated to the condition of the uninjured bone in intensity of bone formation and thickness. These featured were observed in the second group only by day 100. The histological study showed that the structure of the femoral condyle articular cartilage was regular in the first group but its changes were reduced thickness and reduced volumetric density of chondrocytes that might result due to disturbed trophical condition. In the second group the articular cartilage fibres tended to disintegrate on the surface that was accompanied by disturbed integrity of the basophil line and penetration of the vessels into the cartilage. Moreover, the cartilage destruction progressed as the experiment continued and chondrocyte proliferation sharply decreased. Synovitis was considerably expressed. The data obtained in the second group proved high risk of developing osteoarthrosis.

Oleuropein or rutin consumption decreases the spontaneous development of osteoarthritis in the Hartley guinea pig

To assess the potential protective effects of three polyphenols oleuropein, rutin and curcumin, on joint ageing and osteoarthritis (OA) development. Sixty 4-week-old Dunkin-Hartley guinea pigs were randomized into four groups and received daily during 31 weeks either standard guinea pig diet (control group) or a standard guinea pig diet enriched with oleuropein (0.025%), rutin (0.5%) or rutin/curcumin (0.5%/0.25%) association. Biomarkers of OA (Coll2-1, Coll2-1NO2, Fib3-1, Fib3-2, ARGS), as well as inflammation prostaglandin E2 (PGE2) were quantified in the serum. Histological assessments of knee cartilage and synovial membrane were performed at week 4 (five young reference guinea pigs) and week 35. At week 35, guinea pigs in the control group spontaneously developed significant cartilage lesions with mild synovial inflammation. The histological scores of cartilage lesions and synovitis were well correlated with the increased level of serum biomarkers. Histologically, all treatments significantly reduced the cartilage degradation score (P<0.01), but only oleuropein significantly decreased the synovial histological score (P<0.05) and serum PGE2 levels (P<0.01) compared to the control group. Coll2-1 was decreased by rutin and the combination of rutin/curcumin, Fib3-1 and Fib3-2 were only decreased by the rutin/curcumin mixture, while Coll2-1NO2 was significantly decreased by all treatments (P<0.05). Oleuropein and rutin±curcumin significantly slowed down the progression of spontaneous OA lesions in guinea pigs. While no additive effect was seen in the curcumin+rutin group, the differential effects of oleuropein and rutin on inflammatory and cartilage catabolic markers suggest an interesting combination for future studies in OA protection.

Low-intensity pulsed ultrasound activates integrin-mediated mechanotransduction pathway in synovial cells.

Low-intensity pulsed ultrasound (LIPUS) suppresses synovial hyperplasia and synovial cell proliferation characterized for rheumatoid arthritis, but the molecular mechanisms remain unknown. The purpose of this study was to examine the mechanotransduction pathway via the integrin/mitogen-activated protein kinase (MAPK) pathway in LIPUS exposure on the synovial membrane cells. Rabbit knee synovial membrane cell line, HIG-82, was cultured with or without FAK phosphorylation inhibitor, PF-573228. One hour after stimulation with PF-573228, the cells exposed to LIPUS for 20 min or sham exposure. A possible integrin/MAPK pathway was examined by immunofluorescence and Western blotting analysis with antibodies targeting specific phosphorylation sites on intracellular signaling proteins. LIPUS exposure increased phosphorylation of FAK, JNK, ERK, and p38, but the phosphorylation was inhibited by PF-573228. In conclusion, LIPUS exposure might be involved in cell apoptosis and survival of synovial membrane cells via integrin/FAK/MAPK pathway.

Histomorphometric analysis of knee synovial membrane in dogs undergoing leg lengthening by classic Ilizarov method and rapid automatic distraction

This pilot study aimed to test the theory that different lengthening methods affect the microscopic structure of knee joint synovium in diverse ways. This was a descriptive and analytical cross-sectional study of synovium carried out in two experimental models of canine leg lengthening using the Ilizarov fixator. Group 1 (n = 6) used manual gradual distraction most commonly used in clinical practice at one millimetre/day divided into four equal increments, 0.25 mm at each increment. Group 2 (n = 7) used an increased rate of automatic distraction at three millimetres/day divided in 120 increments, 0.025 mm at each increment. At the end of distraction and after fixator removal, the animals were euthanised. Control was via intact dogs. The thickness of the synovial lining layer, number of synovial cell rows, and numerical density of subsynovial microvessels were assessed in digital images for semiautomated computerised analysis of semi-thin sections stained with toluidine blue and methylene blue-basic fuchsin. Comparison of synovitis manifestation was made with grading scale. The vascular and nerve changes in the subsynovial layer were also compared. Group 1 developed marked synovitis, synovium hypervascularisation, degeneration of the nerve fibres in subsynovial nerves with the tendency to regeneration. Group 2 had moderate to mild degree of synovitis with progressive degenerative changes in subsynovial vessels and nerves. Both methods used are unfavourable for the state of the joint synovium, but modify it in different ways.

HISTOMORPHOMETRIC ANALYSIS OF THE KNEE ARTICULAR CARTILAGE AND SYNOVIUM FOR METADIAPHYSEAL LEG LENGTHENING (EXPERIMENTAL-AND-MORPHOLOGICAL STUDY)

The knee articular cartilage and synovial membrane have been studied for metadiaphyseal leg lengthening using the methods of light miscroscopy, computer morpho- and stereometry. The manner of bone integrity breaking, the rate and rhythm of distraction conformed to the lengthening technique most often used in the clinic. The results of the histomorphometric analysis have demonstrated that when osteotomy at the level of metadiaphysis and manual distraction by 1 mm a day for 4 times is performed, synovitis of mild and moderate degree develops through subsynovial layer hypervascularization, as well as reactive-destructive changes in nerve fibers with the tendency to regeneration. The structural-functional changes of reactive and/or destructive-reparative character have been revealed in the articular cartilage, and the manifestation degree of these changes correlates with synovial membrane changes. The intensity of the destructive-reparative processes in the articular cartilage and synovial membrane depends on fixation stability.

Ganghwaljetongyeum, an anti-arthritic remedy, attenuates synoviocyte proliferation and reduces the production of proinflammatory mediators in macrophages: the therapeutic effect of GHJTY on rheumatoid arthritis

BackgroundGanghwaljetongyeum (GHJTY), a complex herbal decoction, is used to treat rheumatoid arthritis. However, the action mechanism of GHJTY is not still unclear on rheumatoid arthritis. In this study, we examined the beneficial effects and the action mechanisms of GHJTY on synoviocyte proliferation and inflammatory mediators.MethodsTo test the effect of GHJTY on synoviocyte proliferation, HIG-82 cells, rabbit knee synovial membrane cells, were treated with GHJTY under IL-1β. To evaluate the effects of GHJTY on proinflammatory mediators, we tested cytokine levels in RAW264.7 cells.ResultsProliferation of HIG-82 cells was significantly inhibited by GHJTY treatment. We found that GHJTY caused cytoskeleton damage to HIG-82 cells. In contrast, treatment of GHJTY did not show any cytotoxicity to other different origin cell lines, HeLa and RAW264.7 cells. GHJTY inhibited IL-1β-mediated NF-κB activation in HIG-82 cells and reduced the LPS-mediated production of proinflammatory cytokines, TNF-α, IL-12, and NO in RAW264.7 cells. In addition, the expression of cyclooxygenase in LPS-activated RAW264.7 cells was also decreased by GHJTY treatment.ConclusionsThese results suggest that GHJTY might effectively attenuate rheumatoid arthritis by inhibiting the production of proinflammatory mediators and the proliferation of synoviocytes.

Interleukin-18-induced inflammatory responses in synoviocytes and chondrocytes from osteoarthritic patients

The major pathological changes of osteoarthritis (OA) include cartilage degeneration and synovial inflammation. Previous studies confirmed that interleukin-1 (IL-1) stimulates the secretion of multiple inflammatory factors in synoviocytes and chondrocytes. IL-18 is a member of the IL-1 superfamily. In this study, the pro-inflammatory effects of IL-18 on synoviocytes and chondrocytes in patients with OA were investigated. Knee synovial membrane and cartilage samples were obtained from OA patients, then primary cells were cultured. Synoviocytes and primary chondrocytes at different generations (primary, secondary and tertiary), were stimulated with IL-18, then inflammatory marker levels, including tumor necrosis factor-α (TNF-α), prostaglandin E2 (PGE₂) and cyclooxygenase-2 (COX-2), were measured using reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay. IL-1 receptor antagonist (IL-1Ra) was applied to interfere with the IL-18 stimulation of chondrocytes, and then the COX-2 expression in chondrocytes and the PGE2 levels in the medium were measured. The expression of IL-18 receptor α (IL-18Rα) and IL-18 receptor β (IL-18Rβ) in synoviocytes and chondrocytes was assessed, using RT-PCR. Our results showed that IL-18 stimulated the COX-2 and TNF-α expressions in primary synoviocytes, while increasing PGE₂ and TNF-α levels in the supernatant (P<0.05) of the culture medium in primary synoviocytes. IL-18 also induced high PGE₂ level production in second-generation synoviocytes (P<0.05). Moreover, IL-18 upregulated COX-2 and TNF-α mRNA in chondrocytes, while promoting PGE₂ and TNF-α (P<0.05) secretions in a dose-dependent manner. The induced effects were not attenuated by the addition of IL-1Ra (P<0.05). IL-18Rα was expressed in the chondrocytes and synoviocytes of 4/8 patients, while IL-18Rβ was expressed in the chondrocytes of 4/8 patients and in the synoviocytes of 2/8 patients. We conclude that IL-18 induces inflammatory responses in synoviocytes and chondrocytes and that this effect was correlated with, although not entirely dependent on, IL-1β.

Low-Intensity Pulsed Ultrasound Reduces the Inflammatory Activity of Synovitis

The purpose of this study was to examine the effect of low-intensity pulsed ultrasound (LIPUS) on the cell proliferation and growth of synovial membrane cells stimulated with inflammatory cytokines, and to evaluate the effectiveness of LIPUS treatment of synovitis in the knee joints of animal models for rheumatoid arthritis. The rabbit knee synovial membrane cell line, HIG-82, was cultured in medium with or without IL-1β or TNF-α. Four hours after stimulation with the cytokines, the cells received LIPUS or sham exposure. Cell proliferation and growth were then analyzed. Using MRL/lpr mice, the anti-inflammatory effects of LIPUS were also evaluated in vivo. Stimulation with proinflammatory cytokines significantly up-regulated cell proliferation which was significantly down-regulated by LIPUS exposure. In MRL/lpr mice, exposure of knee joints to LIPUS caused a significant reduction of histological damage compared to the control. Histological lesions were significantly reduced in the joints treated with LIPUS for 3 weeks. Cox-2-positive cells in the knee joints treated with LIPUS were markedly decreased compared to the control joints. Therefore, LIPUS stimulation may be a medical treatment for joint inflammatory diseases, such as synovitis.

Effects of Low-Intensity Pulsed Ultrasound on the Expression and Activity of Hyaluronan Synthase and Hyaluronidase in IL-1β-Stimulated Synovial Cells

The purpose of this study is to examine effects of low-intensity pulsed ultrasound (LIPUS) on metabolism of hyaluronan (HA) in synovial membrane cells stimulated by IL-1β. Rabbit knee synovial membrane cell line, HIG-82, was cultured in medium with the presence or absence of 1 ng/mL IL-1β, and after 4 h the cell was exposed to LIPUS for 15 min. The mRNA levels of HA synthase (HAS) 2,3, hyaluronidase (HYAL) 2, and cyclooxygenase (COX)-2 were examined by real-time PCR analysis. Concentrations of HA and PGE₂ were quantified by use of enzyme linked immunosorbent assay (ELISA). The COX-2 level was analyzed by western blotting. Gene levels of HAS2 and HAS3 in IL-1β-stimulated cells were up-regulated significantly (p < 0.01) by LIPUS. HYAL2 mRNA was up-regulated by the treatment with IL-1β, whereas down-regulated significantly (p < 0.01) by the following LIPUS exposure. Furthermore, IL-1β stimulation enhanced COX-2 and PGE₂ expression as compared to the untreated control, and IL-1β-induced COX-2 and PGE₂ expression was inhibited by LIPUS. These results suggest that LIPUS enhanced HA synthesis and inhibited HYAL2 expression, leading to the accumulation of high-molecular weight HA. Therefore, LIPUS stimulation may be a better candidate as medical remedy to treat inflammatory joint diseases accompanied with HA degradation in synovial fluid.

Combination Therapy With Montelukast and Ketotifen for Arthritis and Rash Resulting From Idiopathic Hypereosinophilic Syndrome

Idiopathic hypereosinophilic syndrome (IHES) is a rare condition of uncertain etiology characterized by marked peripheral blood eosinophilia and organ system dysfunction that cannot be explained by any factor other than the presence of eosinophils or their potentially toxic products. Diagnostic criteria include 1) a sustained eosinophilia greater than 1500/mm for longer than 6 months, 2) absence of other causes of eosinophilia, including parasitic infections and allergic diseases, and 3) multiorgan involvement (ie, lungs, heart, central nervous system, skin, liver, joints). Steroids represent the initial therapeutic approach, although for those patients unresponsive to steroids, cytotoxic chemotherapy should be considered. We describe a case of IHES characterized by polyarticular inflammatory joint involvement, hypereosinophilia, and urticarioid skin manifestations without visceral involvement. Synovial fluid smears as well as pathology of skin lesions and knee synovial membrane confirmed the presence of numerous eosinophils. The patient was successfully treated with a combination therapy of a cysteinyl leukotriene receptor antagonist (montelukast) and ketotifen. Hypereosinophilic syndrome like in this patient is a rare cause of polyarthritis.