Normal Synovium Research Articles

Histological synovitis score in juvenile idiopathic arthritis and other pediatric synovial inflammatory conditions

ObjectiveThe Krenn’s scoring is a system for histopathological grading of synovial inflammation, and in adults, useful in etiological grouping. The score has only rarely been used in pediatric samples. We aimed to assess the performance of the score in juvenile idiopathic arthritis and other pediatric synovial inflammatory processes in categorization of the disease and in finding characteristic pathological features. DesignWe collected an unselected series of pediatric (age < 16 years) routine synovial biopsy samples to represent normal synovium and inflammatory conditions. The final diagnosis based on clinical follow-up was determined and classified as normal synovium, and different groups according to etiology. Total of 142 patients were analyzed. According to the score, case was classified to normal, low- or high-grade synovitis. ResultsThe synovitis scores in clinically normal synovium were low with 48 % of cases with scores of low-grade synovitis. In structural joint disorders scores varied from normal to low grade synovitis with occasional cases of high grade synovitis. In transient/reactive arthritis scores showed increase, majority clustering to low grade synovitis. In JIA and in bacterial synovitis the scores were higher than in the other groups high grade synovitis being the dominant grade. Extended oligoarthritis showed higher score than persistent oligoarthritis. ROC analysis indicated that JIA could be differentiated from other conditions. ConclusionsThe Krenn’s synovitis score is useful in the etiological classification of pediatric synovial samples, high Krenn’s score suggesting JIA. Observed differences between the subcategories of oligoarthritis may be useful in subclassifying these types of JIA.

New Classification of Rheumatoid Arthritis Based on Immune Cells and Clinical Characteristics.

Rheumatoid arthritis (RA) is a chronic systemic immune disease characterized by joint synovitis, but there are differences in clinical manifestations and serum test results among different patients. This is a bioinformatics study. We first obtained the gene expression profile of RA and normal synovium from the database, and screened the differentially expressed immune related genes for enrichment analysis. Subsequently, we classified RA into three subtypes by unsupervised clustering of serum gene expression profiles based on immune enrichment scores. Then, the enrichment and clinical characteristics of different subtypes were analyzed. Finally, according to the infiltration of different subtypes of immune cells, diagnostic markers were screened and verified by qRT-PCR. C1 subtype is related to the increase of neutrophils, C-reactive protein and erythrocyte sedimentation rate, and joint pain is more significant in patients. C2 subtype is related to the expression of CD8+T cells and Tregs, and patients have mild joint pain symptoms. The RF value of C3 subtype is higher, and the expression of various immune cells is increased. CD4 T cells, NK cells activated, macrophages M1 and neutrophils are immune cells significantly infiltrated in synovium and serum of RA patients. IFNGR1, TRAC, IFITM1 can be used as diagnostic markers of different subtypes. In this study, RA patients were divided into different immune molecular subtypes based on gene expression profile, and immune diagnostic markers were screened, which provided a new idea for the diagnosis and treatment of RA.

Synovial-tissue resident macrophages play proinflammatory functions in the pathogenesis of RA while maintaining the phenotypes in the steady state

Synovial tissue-resident macrophages (STRMs) maintain normal joint homeostasis in a steady state. However, it is unclear whether STRMs still play homeostatic roles or change the functions in the joint of rheumatoid arthritis (RA), where infiltrating peripheral blood monocyte-derived macrophages (PBMoMs) play proinflammatory roles. In the present study, we examined changes in the phenotypes and functions of STRMs in response to RA-related stimuli in vitro. STRMs were prepared from non-inflammatory osteoarthritis (OA) joint synovium, which is histologically indistinguishable from normal joint synovium. PBMoMs were prepared and used for comparison. After stimulation with plate-bound IgG, which mimics anti-citrullinated protein antibody immunocomplex formed in RA joints, or with combinations of RA-related inflammatory mediators, namely tumor necrosis factor-α (TNF-α) and prostaglandin E2 or interferon-γ, PBMoMs downregulated surface markers and genes associated with anti-inflammatory macrophages, and upregulated cytokine and marker genes of proinflammatory macrophages in RA. On the other hand, STRMs hardly changed the expression of surface molecules and marker genes but altered the pattern of cytokine gene expression after stimulation like PBMoMs. Furthermore, in vitro stimulated STRMs promote proinflammatory functions of cocultured synovial fibroblasts. Thus, STRMs might play proinflammatory roles in RA joints, while maintaining their phenotypes in the steady state.

Continuous detrimental activity of intra-articular fibrous scar tissue in correlation with posttraumatic ankle osteoarthritis

Posttraumatic osteoarthritis is primarily characterized by articular cartilage destruction secondary to trauma or fracture events. Even while intra-articular scar tissue can be observed following ankle fractures, little is known about its nature and molecular events linking its biological activity and cartilage deterioration. Here, we investigated scar tissue's histological and molecular characteristics, and its relationship with localized articular cartilage alterations consistent with early osteoarthritic degeneration. Intra-articular scar tissues from sixty-two patients who underwent open reduction internal fixation for ankle fracture were obtained at hardware removal time (6–44 months after fracture). Histological analysis demonstrated that scar tissue has the nature of fibrosis with fibrous tissue hyperplasia, fibroblast proliferation, and chondrometaplasia. These fibrous scar tissues showed overexpressed pro-inflammatory cytokines and high mRNA expression levels of osteoarthritis-related markers (cytokines, chemokines, and enzymes) compared to the normal synovium. Furthermore, those transcriptional levels were significantly correlated with the grade of talar chondral degeneration. Our findings suggest that following an ankle fracture, the intra-articular fibrous scar tissue exhibits high catabolic and inflammatory activity, which has a long-lasting negative impact correlated to cartilage deterioration in the development of posttraumatic osteoarthritis.

Synovium is a sensitive tissue for mapping the negative effects of systemic iron overload in osteoarthritis: identification and validation of two potential targets

BackgroundThe prevention and treatment of osteoarthritis (OA) pose a major challenge in its research. The synovium is a critical tissue in the systematic treatment of OA. The present study aimed to investigate potential target genes and their correlation with iron overload in OA patients.MethodsThe internal datasets for analysis included the microarray datasets GSE46750, GSE55457, and GSE56409, while the external datasets for validation included GSE12021 and GSE55235. The GSE176308 dataset was used to generate single-cell RNA sequencing profiles. To investigate the expression of the target genes in synovial samples, quantitative reverse transcription-PCR, western blotting, and immunohistochemical assay were conducted. ELISA was used to detect the levels of ferritin and Fe2+ in both serum and synovium.ResultsJUN and ZFP36 were screened from the differentially expressed genes, and their mRNA were significantly reduced in the OA synovium compared to that in normal synovium. Subsequently, complex and dynamically evolving cellular components were observed in the OA synovium. The mRNA level of JUN and ZFP36 differed across various cell clusters of OA synovium and correlated with immune cell infiltration. Moreover, ferritin and Fe2+ were significantly increased in the serum and synovium of OA patients. Further, we found that JUN elevated and ZFP36 decreased at protein level.ConclusionsThe synovium is a sensitive tissue for mapping the adverse effects of systemic iron overload in OA. JUN and ZFP36 represent potential target genes for attenuating iron overload during OA treatment. Some discrepancies between the transcription and protein levels of JUN suggest that post-transcriptional modifications may be implicated. Future studies should also focus on the roles of JUN and ZFP36 in inducing changes in cellular components in the synovium during OA pathogenesis.

E71 Distribution of cells and extracellular matrix in the normal marginal synovium of the hip joint capsule during the postnatal period

Abstract Introduction The marginal transitional zone is of fundamental importance in joint pathology. It includes the marginal cartilage and marginal synovium. The normal marginal synovium takes part in articular cartilage repair (Adachi et al. 2012). The inflamed marginal synovium, in turn, promotes irreversible damage to the adjacent cartilage and bone (Hitchon &amp; El-Gabalawy, 2011). Objectives The morphology of the normal marginal synovium from birth till the 90th day was studied using the hip joints of Wistar rats. Methods The collection of joints was carried out on the first, seventh, 14th, 30th, 45th, 60th and 90th day (six rats in each group, 42 in total). Joints were fixed, decalcified and dehydrated. Histologic paraffine sections were stained with Mallory's trichrome and Hart's elastin staining, analyzed at the acetabular labrum by light microscopy (χ100), morphometrical and statistical (P &amp;lt; 0.05) methods with extrapolation to humans (Sengupta, 2013). The obtained data were presented as percentages. Results In the marginal synovium, cells and components of the extracellular matrix, which are typical for fibrous connective tissue, could be distinguished. The synovial lining cells are embedded in the modified basement membrane which is a meshwork of collagen and elastic fibers. They form a three-dimensional reticulum in it and adjacent synovial subintima. Vessels of the microcirculatory bed are also widely represented in the marginal synovium. The overall quantity of collagen fibers increased gradually, exceeding on the 90th day (∼18–19 human years) that of newborns by one and a half times (48.92 ± 3.39 vs 72.17 ± 7.77; P = 0.023). The number of arranged collagen fibers increased gradually, exceeding on the 90th day that of newborns almost twofold (37.25 ± 3.21 vs 60.08 ± 5.99; P = 0.008). The area of distribution of disarranged collagen fibers in newborns was almost four and a half times less than the overall quantity of collagen fibers (11.67 ± 1.03 vs 48.92 ± 3.39; P &amp;lt; 0.001) and did not change substantially throughout the experiment. The number of elastic fibers increased till the 30th day (0.63 ± 0.11 vs 18.3 ± 1.25; P &amp;lt; 0.001) and did not undergo significant changes later. In newborns, the area of distribution of ground substance was not considerably higher than the ratio of disarranged collagen fibers (10.42 ± 1.8 vs 11.67 ± 1.03; P = 0.562) and did not undergo substantial changes later. The distribution density of vessels increased till the 30th-45th day (4.5 ± 0.67 vs 7.5 ± 0.82 and 7.7 ± 0.54; P &amp;lt; 0.001; ∼9-14th human years), followed by a gradual decrease from the 60th day (7.05 ± 0.72; ∼16 human years). The maximum number of cells was in newborns. Subsequently, it gradually decreased and was almost three times less on the 90th day than that of newborns (38.01 ± 3.06 vs 12.58 ± 1.53; P &amp;lt; 0.001). Conclusion The morphogenesis of the marginal synovium during the postnatal period is accompanied by a gradual decrease in the number of cells, an increase in the area occupied by elastic and arranged collagen fibers with a relatively constant distribution density of disarranged collagen fibers and ground substance together with an undulatory pattern in the distribution of vessels. These peculiarities emphasize the morphological uniqueness of the marginal transitional zone, which forms the innate nonspecific barrier between the joint capsule and the articular cartilage, and thereby provides the necessary anatomical and physiological integrity of the joint as a whole organ. Knowledge about the patterns of morphogenesis of the normal marginal synovium will help to understand joint pathology. Ethics The study was approved by the ethics committee of the Zaporizhzhia State Medical University.

E72 Distribution of cells and extracellular matrix in the marginal synovium during the postnatal period

Abstract Introduction The marginal transitional zone of the joint is of fundamental importance in joint pathology. The normal marginal synovium takes part in articular cartilage repair (Adachi et al. 2012). The inflamed marginal synovium, in turn, promotes irreversible damage to the adjacent cartilage and bone (Hitchon &amp; El-Gabalawy, 2011). Early erosive damage in rheumatoid arthritis and osteophyte formation in osteoarthritis occurs primarily here (Allard et al. 1990). Objectives The morphology of the marginal synovium from birth till the 90th day was studied using the hip joints of Wistar rats. Methods The collection of joints was carried out on the first, seventh, 14th, 30th, 45th, 60th and 90th day (six rats in each group, 42 in total). Joints were fixed, decalcified and dehydrated. Histologic paraffine sections were stained with Mallory's trichrome and Hart's elastin staining, analyzed at the acetabular labrum by light microscopy (χ100), morphometrical and statistical (P &amp;lt; 0.05) methods with extrapolation to humans (Sengupta, 2013). The obtained data were presented as percentages. Results In the marginal synovium, cells and components of the extracellular matrix, which are typical for fibrous connective tissue, could be seen. The synovial lining cells are embedded in the modified basement membrane which includes collagen and elastic fibers. They form a three-dimensional reticulum in it and adjacent synovial subintima. Vessels of the microcirculatory bed are also widely represented in the marginal synovium. The overall quantity of collagen fibers increased gradually, exceeding on the 90th day (approximately corresponds to 18–19 human years) that of newborns by one and a half times (48.92 ± 3.39 vs 72.17 ± 7.77; P = 0.023). The number of arranged collagen fibers increased gradually, exceeding on the 90th day that of newborns almost twofold (37.25 ± 3.21 vs 60.08 ± 5.99; P = 0.008). The area of distribution of disarranged collagen fibers in newborns was almost four and a half times less than the overall quantity of collagen fibers (11.67 ± 1.03 vs 48.92 ± 3.39; P &amp;lt; 0.001) and did not change substantially throughout the experiment. The number of elastic fibers increased till the 30th day (0.63 ± 0.11 vs 18.3 ± 1.25; P &amp;lt; 0.001) and did not undergo significant changes later. In newborns, the area of distribution of ground substance was not considerably higher than the ratio of disarranged collagen fibers (10.42 ± 1.8 vs 11.67 ± 1.03; P = 0.562) and did not undergo substantial changes later. The distribution density of vessels increased till the 30th-45th day (4.5 ± 0.67 vs 7.5 ± 0.82 and 7.7 ± 0.54; P &amp;lt; 0.001; approximately corresponds to the 9th and 14th human years respectively), followed by a gradual decrease from the 60th day (7.05 ± 0.72; roughly corresponds to the 16th human years). The maximum number of cells was in newborns. Subsequently, it gradually decreased and was almost three times less on the 90th day than that of newborns (38.01 ± 3.06 vs 12.58 ± 1.53; P &amp;lt; 0.001). Conclusion The morphogenesis of the marginal synovium during the postnatal period is accompanied by a gradual decrease in the number of cells, an increase in the area occupied by elastic and arranged collagen fibers with a relatively constant distribution density of disarranged collagen fibers and ground substance together with an undulatory pattern in the distribution of vessels. These peculiarities emphasize the morphological uniqueness of the marginal transitional zone, which forms the innate nonspecific barrier between the joint capsule and the articular cartilage, and thereby provides the necessary anatomical and physiological integrity of the joint as a whole organ. Knowledge about the patterns of morphogenesis of the marginal synovium will help in the future to analyze the modeled pathological or boundary states in it. Ethics The study was approved by the ethics committee of the Zaporizhzhia State Medical University.

Nesfatin-1 stimulates BMP5 expression and osteoclastogenesis in rheumatoid arthritis

Rheumatoid arthritis (RA) is a common autoimmune disease marked by immune cell activation and chronic inflammation in the synovium accompanied by osteoclast activation and local joint destruction. Increased levels of the adipokine nesfatin-1 in RA synovium are associated with proinflammatory cytokines. Our analysis of datasets from the Gene Expression Omnibus (GEO) database and synovial tissue samples from RA patients revealed that these had higher levels of nesfatin-1 and osteoclast markers compared with normal synovium. These findings were the same in tissue samples from mice with collagen-induced arthritis (CIA) and normal healthy controls. RNA sequencing analysis revealed that nesfatin-1 increased levels of bone morphogenetic protein-5 (BMP5) expression via JAK/STAT signaling in RA synovial fibroblasts. Finally, we found that nesfatin-1 short hairpin RNA reduced BMP5 and osteoclast formation in CIA mice. These findings provide new insights into the pathogenesis of RA.

Identification of aging-related biomarkers and immune infiltration characteristics in osteoarthritis based on bioinformatics analysis and machine learning.

Osteoarthritis (OA) is a degenerative disease closely related to aging. Nevertheless, the role and mechanisms of aging in osteoarthritis remain unclear. This study aims to identify potential aging-related biomarkers in OA and to explore the role and mechanisms of aging-related genes and the immune microenvironment in OA synovial tissue. Normal and OA synovial gene expression profile microarrays were obtained from the Gene Expression Omnibus (GEO) database and aging-related genes (ARGs) from the Human Aging Genomic Resources database (HAGR). Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Disease Ontology (DO), and Gene set variation analysis (GSVA) enrichment analysis were used to uncover the underlying mechanisms. To identify Hub ARDEGs with highly correlated OA features (Hub OA-ARDEGs), Weighted Gene Co-expression Network Analysis (WGCNA) and machine learning methods were used. Furthermore, we created diagnostic nomograms and receiver operating characteristic curves (ROC) to assess Hub OA-ARDEGs' ability to diagnose OA and predict which miRNAs and TFs they might act on. The Single sample gene set enrichment analysis (ssGSEA) algorithm was applied to look at the immune infiltration characteristics of OA and their relationship with Hub OA-ARDEGs. We discovered 87 ARDEGs in normal and OA synovium samples. According to functional enrichment, ARDEGs are primarily associated with inflammatory regulation, cellular stress response, cell cycle regulation, and transcriptional regulation. Hub OA-ARDEGs with excellent OA diagnostic ability were identified as MCL1, SIK1, JUND, NFKBIA, and JUN. Wilcox test showed that Hub OA-ARDEGs were all significantly downregulated in OA and were validated in the validation set and by qRT-PCR. Using the ssGSEA algorithm, we discovered that 15 types of immune cell infiltration and six types of immune cell activation were significantly increased in OA synovial samples and well correlated with Hub OA-ARDEGs. Synovial aging may promote the progression of OA by inducing immune inflammation. MCL1, SIK1, JUND, NFKBIA, and JUN can be used as novel diagnostic biomolecular markers and potential therapeutic targets for OA.

In major joint diseases the human synovium retains its potential to form repair cartilage

The inner surface layer of human joints, the synovium, is a source of stem cells for the repair of articular cartilage defects. We investigated the potential of the normal human synovium to form novel cartilage and compared its chondrogenic capacity with that of two patient groups suffering from major joint diseases: young adults with femoro-acetabular impingement syndromes of the hip (FAI), and elderly individuals with osteoarthritic degeneration of the knee (OA). Synovial membrane explants of these three patient groups were induced in vitro to undergo chondrogenesis by growth factors: bone morphogenetic protein-2 (BMP-2) alone, transforming growth factor-β1 (TGF-β1) alone, or a combination of these two. Quantitative evaluations of the newly formed cartilages were performed respecting their gene activities, as well as the histochemical, immunhistochemical, morphological and histomorphometrical characteristics. Formation of adult articular-like cartilage was induced by the BMP-2/TGF-β1 combination within all three groups, and was confirmed by adequate gene-expression levels of the anabolic chondrogenic markers; the levels of the catabolic markers remained low. Our data reveal that the chondrogenic potential of the normal human synovium remains uncompromised, both in FAI and OA. The potential of synovium-based clinical repair of joint cartilage may thus not be impaired by age-related joint pathologies.

Distinct macrophage polarization in acute and chronic gout

AbstractMacrophage polarization mediates the development of inflammatory diseases. However, the polarization status at various stages of gout is not fully understood. Our study aimed to define the evolution of macrophage polarization in acute and chronic gout. Normal human synovium and synovium with tophi were collected for immunofluorescence (IF). Rat gouty joints were collected for joint thickness assessment and pathological evaluation. Tissue mRNA expression of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) were evaluated. Mouse peritoneal macrophages and THP-1 derived macrophages were stimulated by monosodium urate (MSU) crystals and were collected for detection of interleukin (IL) -1β and IL-37 levels and iNOS/Arg-1 ratio. Arg-1 and IL-37 were highly expressed in normal synovium and synovium with tophi. In rat gouty joints, the inflammatory cell counts and ankle thickness began to increase at 2 h, peaked at 24 h, and was decreased spontaneously. An increase in macrophages preceded the neutrophils infiltration. Infiltration of M1 was positively related with the severity of arthritis. M2 appeared in an early stage (at 2 h) of inflammation. The number of M1 macrophages was comparable to that of M2 from 2 to 12 h and exceeded M2 number at 18 h and 24 h. The ratios of M2/M1 reversed at 48 h and remained reversed until 120 h. In mice gouty joints, iNOS/Arg-1 mRNA ratio was significantly higher than the that in control group at 8 h. The proportion of neutrophils and M1-macrophages reached peak at 4 h in mice model with peritoneal gout. Concentration of IL-1β and ratio of iNOS/Arg-1 were increased at 6 h, peaked at 48 h, and were then decreased at 72 h in vitro, while the concentration of IL-37 peaked at 2 h and then decreased. In summary, altered macrophage polarization was observed in various stages of gouty inflammation. Macrophages in acute gout were polarized into M1 at early stage and into M2 at later stage while the macrophages in chronic gout mainly were only polarized towards M2. The number of M1 rose with the progression of inflammation. Early increase of M2 was observed, which might be generated directly from M0.

Synovial membrane-derived mesenchymal progenitor cells from osteoarthritic joints in dogs possess lower chondrogenic-, and higher osteogenic capacity compared to normal joints

BackgroundSynovial membrane-derived mesenchymal progenitor cells (SM-MPCs) are a promising candidate for the cell-based treatment of osteoarthritis (OA) considering their in vitro and in vivo capacity for cartilage repair. However, the OA environment may adversely impact their regenerative capacity. There are no studies for canine (c)SM-MPCs that compare normal to OA SM-MPCs, even though dogs are considered a relevant animal model for OA. Therefore, this study compared cSM-MPCs from normal and OA synovial membrane tissue to elucidate the effect of the OA environment on MPC numbers, indicated by CD marker profile and colony-forming unit (CFU) capacity, and the impact of the OA niche on tri-lineage differentiation.MethodsNormal and OA synovial membrane were collected from the knee joints of healthy dogs and dogs with rupture of the cruciate ligaments. The synovium was assessed by histopathological OARSI scoring and by RT-qPCR for inflammation/synovitis-related markers. The presence of cSM-MPCs in the native tissue was further characterized with flow cytometry, RT-qPCR, and immunohistochemistry, using the MPC markers; CD90, CD73, CD44, CD271, and CD34. Furthermore, cells isolated upon enzymatic digestion were characterized by CFU capacity, and a population doublings assay. cSM-MPCs were selected based on plastic adherence, expanded to passage 2, and evaluated for the expression of MPC-related surface markers and tri-lineage differentiation capacity.ResultsSynovial tissue collected from the OA joints had a significantly higher OARSI score compared to normal joints, and significantly upregulated inflammation/synovitis markers S100A8/9, IL6, IL8, and CCL2. Both normal and OA synovial membrane contained cells displaying MPC properties, including a fibroblast-like morphology, CFU capacity, and maintained MPC marker expression over time during expansion. However, OA cSM-MPCs were unable to differentiate towards the chondrogenic lineage and had low adipogenic capacity in contrast to normal cSM-MPCs, whereas they possessed a higher osteogenic capacity. Furthermore, the OA synovial membrane contained significantly lower percentages of CD90+, CD44+, CD34+, and CD271+ cells.ConclusionsThe OA environment had adverse effects on the regenerative potential of cSM-MPCs, corroborated by decreased CFU, population doubling, and chondrogenic capacity compared to normal cSM-MPCs. OA cSM-MPCs may be a less optimal candidate for the cell-based treatment of OA than normal cSM-MPCs.

Role of magnetic resonance imaging in assessment of extensor compartment abnormalities of knee joint – A cross-sectional study

INTRODUCTION: The patellofemoral joint (PFJ) is a complex joint structure with high functional and biomechanical requirements. The magnetic resonance imaging (MRI) is an effective tool for definition and characterization of knee pathology as it can accurately detect, localize, and characterize various pathologies of the knee joint. OBJECTIVES: (1) Evaluate MR images to identify different pathologies that are seen in the extensor compartment of the knee joint. (2) Quantitative assessment of patellofemoral measurements to determine predisposing risk factors that contribute to patellofemoral joint instability. METHODOLOGY: A cross-sectional study was conducted from January 2017 to December 2017 with a sample size of 30 in the Department of Radiodiagnosis at The KLE's Dr. Prabhakar Kore Hospital and MRC, Belagavi. The data were collected using a pretested semi-structured questionnaire. The pro forma included sociodemographic profile of the study subjects, clinical presentation MRI to identify different pathologies that are seen in extensor compartment of the knee joint and quantitative assessment of patellofemoral measurements to determine predisposing risk factors that contribute to patellofemoral joint instability. RESULTS: Among the study population majority of people had normal synovium, laterally subluxated/dislocated patella. Among the study population, the majority had normal lateral trochlear inclination angle, normal trochlear facet asymmetry, normal trochlear depth, normal sulcus angle, and trochlear dysplasia. Twenty-four (80%) people had normal Insall–Salvati index, 27 (90%) people had normal lateral patella femoral angle, 25 (83.30%) people had normal lateralization of the patella, 28 (93.30%) people had normal tibial tubercle to trochlear groove distance, majority, i.e., 11 (36.70%) had Hoffa's fat pad edema followed by chondromalacia patella, trochlear dysplasia, medial patellofemoral ligament tear, patella alta, synovial plicae, subluxated/dislocated patella, transient dislocated patella, Osgood–Schlatter disease, and patellar tendonitis. CONCLUSION: MRI has also been useful in identifying extensor compartment pathologies that lead to anterior knee pain.

Identification of SCRG1 as a Potential Therapeutic Target for Human Synovial Inflammation.

Synovial inflammation of joint tissue is the most important cause of tissue damage, joint destruction, and disability and is associated with higher morbidity or mortality. Therefore, this study aims to identify key genes in osteoarthritis synovitis tissue to increase our understanding of the underlying mechanisms of osteoarthritis and identify new therapeutic targets. Five GEO datasets with a total of 41 normal synovial membrane tissues and 45 osteoarthritis synovial membrane samples were used for analysis, and seven common differential genes were identified. The classification model constructed by LASSO analysis showed that six genes including CDKN1A, FOSB, STMN2, SLC2A3, TAC, and SCRG1 can be used as biomarkers of osteoarthritis, and the SCRG1 gene shows importance in osteoarthritis. Furthermore, drug database enrichment found that these six DEGs may be the drug targets of synovitis in osteoarthritis, and Valproic Acid CTD 00006977 may be a potential targeted therapeutic drug of SCRG1. Spearman correlation analysis was performed on the SCRG1 gene, and 27 genes with consistent expression were obtained. Functional analysis showed that 27 genes were mainly involved in metabolism, complement, antigen presentation, apoptosis, and regulation of immune pathways. The co-regulatory network of TFs-miRNA suggested that the SCRG1 gene may be regulated by hsa-miR-363-3p miRNA. In conclusion, SCRG1, as a diagnostic marker of osteoarthritis, co-regulates immune-related pathways through the interaction of related proteins, playing an important role in the occurrence and development of osteoarthritis, which may be a novel drug target.

Pharmacological Investigation of Cassia hirsuta Seed Extract for its Anti-inflammatory Activity in Acute and Chronic Inflammatory Animal Models

Objective The declared goal of this research was to investigate anti-inflammatory and anti-arthritic effectiveness of ethanolic extract of Cassia hirsuta seeds EECH in animals employing Carrageenan-induced paw CIP edema and Complete Freundrsquos adjuvant CFA induced arthritis models.Methodology The seeds of Cassia hirsuta were collected shade dried extracted using ethanol in Soxhlet extractor and the extract was tested for phytochemicals and acute oral toxicity 423 - OECD guidelines. CIP edema was examined to ascertain anti-inflammatory property. The anti-arthritic activity of CFA was analyzed by measuring paw volume arthritic index and body weight. Hematological parameters and markers in serum levels of enzymes like SGPT SGOT serum RF and histopathological studies were also performed.Results The seed extract possessed various phytoconstituents like alkaloids flavonoids terpenoids phenols and tannins as active constituents. In acute oral toxicity study the EECH produced no lethality with maximum dose of up to 2000 mgkg bw. Hence 120th 110th amp 15th mgkg b.w were taken as lower moderate and higher dose respectively. EECH at concentration of 100 200 amp 400 mgkg was able to reduce the inflammation induced by carrageenan in rats. However 200 mgkg 400 mgkg has shown significant inhibition of arthritis in rats when challenged with CFA 0.1 mL. The EECH of dose 200 mgkg 400 mgkg had maintained normal architecture of bone normal areolar synovium and normal histology in comparison to arthritis control.Conclusion According to our findings the EECH demonstrated anti-oxidant anti-inflammatory amp antiarthritic activity in a dose-dependent manner. The phytoconstituents like flavonoids terpenoids and phenols as active constituents in the extract may justify its activity.nbsp

Synergistic upregulation of ADAMTS4 (aggrecanase-1) by cytokines and its suppression in knee osteoarthritic synovial fibroblasts

AbstractThe ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family includes nine members with aggrecan-degrading activity, i.e., ADAMTS1, 4, 5, 8, 9, 15, 16, 18, and 20. However, their systematic expression profile in knee osteoarthritis (OA) synovium and effects of cytokines and growth factors on the expression in OA synovial fibroblasts remain elusive. In this study, expression of all nine aggrecanolytic ADAMTS species was assessed by quantitative real-time PCR in OA and control normal synovial tissues. OA synovial fibroblasts were treated with interleukin-1α (IL-1α), IL-1β, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), vascular endothelial growth factor165, and heparin-binding epidermal growth factor, and analyzed for the expression of the ADAMTS species. The signaling pathways and inhibition of ADAMTS4 expression by high-molecular-weight hyaluronan, adalimumab, tocilizumab, and signaling molecule inhibitors were studied. ADAMTS1, 4, 5, 9, and 16 were expressed in OA synovium, but only ADAMTS4 expression was significantly higher in OA as compared to normal synovium. IL-1α, TNF-α, and TGF-β markedly increased ADAMTS4 expression, while their effects were minimal for the other ADAMTS species. ADAMTS4 was synergistically upregulated by treatment with IL-1α and TNF-α, IL-1α and TGF-β, or IL-1α, TNF-α and TGF-β. The signaling molecules' inhibitors demonstrated that IL-1α-induced ADAMTS4 expression is predominantly through TGF-β-associated kinase 1 (TAK1), and the TNF-α-stimulated expression is via TAK1 and nuclear factor-κB (NF-κB). The TGF-β-promoted expression was through the activin receptor-like kinase 5 (ALK5)/Smad2/3, TAK1, and non-TAK1 pathways. Adalimumab blocked TNF-α-stimulated expression. ADAMTS4 expression co-stimulated with IL-1α, TNF-α and TGF-β was abolished by treatment with adalimumab, TAK1 inhibitor, and ALK5/Smad2/3 inhibitor. These data demonstrate marked and synergistic upregulation of ADAMTS4 by IL-1α, TNF-α and TGF-β in OA synovial fibroblasts, and suggest that concurrent therapy with an anti-TNF-α drug and inhibitor(s) may be useful for prevention against aggrecan degradation in OA.

Identification and Validation of ATF3 Serving as a Potential Biomarker and Correlating With Pharmacotherapy Response and Immune Infiltration Characteristics in Rheumatoid Arthritis.

Background: Although disease-modifying antirheumatic drugs (DMARDs) have significantly improved the prognosis of patients with rheumatoid arthritis (RA), approximately 40% of RA patients have limited response. Therefore, it was essential to explore new biomarkers to improve the therapeutic effects on RA. This study aimed to develop a new biomarker and validate it by an in vitro study. Methods: The RNA-seq and the clinicopathologic data of RA patients were downloaded from Gene Expression Omnibus (GEO) databases. Differentially expressed genes were screened in the GPL96 and GPL570 databases. Then, weighted gene co-expression network analysis (WGCNA) was used to explore the most correlated gene modules to normal and RA synovium in the GPL96 and GPL570 databases. After that, the differentially expressed genes were intersected with the correlated gene modules to find the potential biomarkers. The CIBERSORT tool was applied to investigate the relationship between activated transcription factor 3 (ATF3) expression and the immune cell infiltration, and Gene Set Enrichment Analysis (GSEA) was used to investigate the related signaling pathways of differentially expressed genes in the high and low ATF3 groups. Furthermore, the relationships between ATF3 expression and clinical parameters were also explored in the GEO database. Finally, the role of ATF3 was verified by in vitro cell experiments. Results: We intersected the differentially expressed genes and the most correlated gene modules in the GPL570 and GPL96 databases and identified that ATF3 is a significant potential biomarker and correlates with some clinical–pharmacological variables. Immune infiltration analysis showed that activated mast cells had a significant infiltration in the high ATF3 group in the two databases. GSEA showed that metabolism-associated pathways belonged to the high ATF3 groups and that inflammation and immunoregulation pathways were enriched in the low ATF3 group. Finally, we validated that ATF3 could promote the proliferation, migration, and invasion of RA fibroblast-like synoviocyte (FLS) and MH7A. Flow cytometry showed that ATF3 expression could decrease the proportion of apoptotic cells and increase the proportion of S and G2/M phase cells. Conclusion: We successfully identified and validated that ATF3 could serve as a novel biomarker in RA, which correlated with pharmacotherapy response and immune cell infiltration.

The protective effects of allopurinol against IL-17A-induced inflammatory response in mast cells

Rheumatoid arthritis (RA) is a common autoimmune disease in the elderly and it has been recently reported to be significantly associated with the activation of mast cells in joint tissues. IL-17A is a vital mediator that stimulates the activation of inflammation. Allopurinol is a classic agent for the suppression of uric acid production, recently reported to exert therapeutic effects on RA. In the present study, we investigated the regulatory effect of allopurinol against IL-17A-induced inflammatory response in mast cells and explored the potential mechanism of allopurinol on RA treatment. Firstly, we found that compared to normal synovium, IL-17A was significantly upregulated in the human RA synovium. IL-17A was used to stimulate an inflammatory state in mast cells in the absence or presence of allopurinol. We found that the production of inflammatory factors, PGE2, and COX-2 was significantly elevated in IL-17A-treated mast cells, accompanied by the activation of the iNOS/NO axis and the elevated secretion of ROS. After treatment with allopurinol, the elevated inflammation, activated COX-2/PGE2 and iNOS/NO axis, and oxidative stress were all dramatically alleviated. Mechanistically, the activated JNK/AP-1 and NF-κB pathways in IL-17A-treated mast cells were dramatically suppressed by the introduction of allopurinol. Taken together, our data reveal that allopurinol significantly alleviated the IL-17A-induced inflammatory response in mast cells.

Metabolic changes in synovial cells in early inflammation: Involvement of CREB phosphorylation in the anti-inflammatory effect of 2-deoxyglucose

The involvement of metabolic reprogramming has been suggested to contribute to the pathophysiology of rheumatoid arthritis (RA). Glycolysis is enhanced in synovial cell metabolism in RA patients. Inhibitors of glycolysis are known to have anti-inflammatory effects. But, changes in the metabolism of normal synovial membranes or synovial cells during the early stages of inflammation remains unknown. Moreover, there are still many aspects of inflammatory signaling pathways altered by glycolysis inhibitors, that remain unclear. In this study we found that, in normal, non-pathological bovine synovial cells, most of ATP synthesis was generated by mitochondrial respiration. However, during the early of stages inflammation, initiated by lipopolysaccharide (LPS) exposure, synovial cells shifted to glycolysis for ATP production. The glycolysis inhibitor 2-deoxyglucose (2DG) reversed LPS induced increases in glycolysis for ATP production and suppressed the expression of inflammatory cytokines and proteolytic enzymes. 2DG suppressed the phosphorylation of the transcription factor cAMP response element binding protein (CREB) enhanced by LPS. Treatment with a CREB inhibitor reversed the expression of LPS-stimulated inflammatory cytokines and proteolytic enzymes. This study showed that changes in metabolism occur during the early stages of inflammation of synovial cells and can be reversed by 2DG and signaling pathways associated with CREB phosphorylation.

Identification of key regulators responsible for dysregulated networks in osteoarthritis by large-scale expression analysis

BackgroundOsteoarthritis (OA) is a worldwide musculoskeletal disorder. However, disease-modifying therapies for OA are not available. Here, we aimed to characterize the molecular signatures of OA and to identify novel therapeutic targets and strategies to improve the treatment of OA.MethodsWe collected genome-wide transcriptome data performed on 132 OA and 74 normal human cartilage or synovium tissues from 7 independent datasets. Differential gene expression analysis and functional enrichment were performed to identify genes and pathways that were dysregulated in OA. The computational drug repurposing method was used to uncover drugs that could be repurposed to treat OA.ResultsWe identified several pathways associated with the development of OA, such as extracellular matrix organization, inflammation, bone development, and ossification. By protein-protein interaction (PPI) network analysis, we prioritized several hub genes, such as JUN, CDKN1A, VEGFA, and FOXO3. Moreover, we repurposed several FDA-approved drugs, such as cardiac glycosides, that could be used in the treatment of OA.ConclusionsWe proposed that the hub genes we identified would play a role in cartilage homeostasis and could be important diagnostic and therapeutic targets. Drugs such as cardiac glycosides provided new possibilities for the treatment of OA.