Clinical Signs Of Arthritis Research Articles

Therapeutic Potential of Olax subscorpioidea Root Extracts in Managing Inflammatory Conditions: A Preclinical Study

Olax subscorpioidea, a plant species endemic to tropical Africa, has long been utilized in traditional medicine for its purported anti-inflammatory and analgesic properties. This preclinical study evaluates the therapeutic potential of Olax subscorpioidea root extracts in managing inflammatory conditions, specifically in a Freund’s adjuvant (CFA)-collagen type II induced arthritis model in albino rats. Both aqueous and ethanol extracts were prepared and assessed for their effects on inflammatory parameters, including cytokine levels, paw edema, joint swelling, and histopathological changes. The study found that both extracts significantly reduced inflammation and clinical signs of arthritis, with the ethanol extract demonstrating a broader range of bioactive compounds and potentially greater efficacy. Mechanistic insights suggest that the extracts exert their effects through inhibition of pro-inflammatory enzymes, modulation of cytokine production, and enhancement of antioxidant activity. Safety and toxicity assessments confirmed the non-toxic nature of the extracts at tested doses. These findings validate the traditional use of Olax subscorpioidea for inflammatory conditions and support its potential integration into modern therapeutic regimens. Future research should focus on dosage optimization, long-term effects, and clinical trials to further establish the efficacy and safety of Olax subscorpioidea root extracts in human subjects. Keywords: Olax subscorpioidea, Root Extracts, Inflammatory Conditions, Preclinical Study.

A novel inhibitor of class IIa histone deacetylases attenuates collagen-induced arthritis.

Most inhibitors of histone deacetylases (HDACs) are not selective and are cytotoxic. Some have anti-inflammatory activity in disease models, but cytotoxicity prevents long-term uses in non-fatal diseases. Inhibitors selective for class IIa HDACs are much less cytotoxic and may have applications in management of chronic inflammatory diseases. LL87 is a novel HDAC inhibitor examined here for HDAC enzyme selectivity. It was also investigated in macrophages for cytotoxicity and for inhibition of lipopolysaccharide (LPS)-stimulated cytokine secretion. In a rat model of collagen-induced arthritis, LL87 was investigated for effects on joint inflammation in Dark Agouti rats. Histological, immunohistochemical, micro-computed tomography and molecular analyses characterise developing arthritis and anti-inflammatory efficacy. LL87 was significantly more inhibitory against class IIa than class I or IIb HDAC enzymes. In macrophages, LL87 was not cytotoxic and reduced both LPS-induced secretion of pro-inflammatory cytokines, and IL6-induced class IIa HDAC activity. In rats, LL87 attenuated paw swelling and clinical signs of arthritis, reducing collagen loss and histological damage in ankle joints. LL87 decreased immune cell infiltration, especially pro-inflammatory macrophages and osteoclasts, into synovial joints and significantly reduced expression of pro-inflammatory cytokines and tissue-degrading proteases. A novel inhibitor of class IIa HDACs has been shown to have an anti-inflammatory and anti-arthritic profile distinct from current therapies. It is efficacious in reducing macrophage infiltration and joint inflammation in a chronic model of rat arthritis.

Modulatory role of prenatal alcohol exposure and adolescent stress on the response to arthritis challenge in adult female rats.

SummaryBackgroundThere are known environmental risk factors associated with rheumatoid arthritis; however, less is known regarding how the prenatal environment impacts later-life risk for rheumatoid arthritis. Based on preliminary clinical data suggesting that individuals with fetal alcohol spectrum disorder (FASD) are at higher risk for autoimmune disorders, this study investigated the modulatory impact of prenatal alcohol exposure (PAE) on the inflammatory disease profile in an adjuvant-induced arthritis rat model.MethodsPregnant rats received liquid ethanol or control diet throughout gestation. To model the increased exposure to stressors often experienced by individuals with FASD, adolescent offspring were exposed to chronic mild stress (CMS) or remained undisturbed. In adulthood, experimental arthritis was initiated and rats terminated either at the peak or following resolution from inflammation to assess endocrine, immune, and histopathological outcomes.FindingsPAE rats had an increased incidence and severity of, and impaired recovery from, arthritis. Increased joint damage was observed in PAE animals, even in the face of apparent recovery from the clinical signs of arthritis, while it appeared that oestradiol may have a protective role. Moreover, with the combination of PAE and adolescent stress, increased macrophage density was detected in the synovium of PAE but not control rats.InterpretationThese findings demonstrate that PAE alters the severity and course of arthritis, highlighting the potential immunomodulatory impact of adverse prenatal exposures. In particular, these data have implications for understanding preliminary data that suggest a heightened propensity for autoimmune disorders in individuals with FASD.

P083 Primary Goujerot- Sjögren Syndrome: a pediatric case report

Abstract Background Gourgerot-Sjögren syndrome (GSS) is a systemic autoimmune disease characterized by lymphocytic infiltration of the exocrine glands leading to dryness of the mucous membranes. It may also involve other organs and organ systems. GSS can be primary (pGSS) or secondary due to other autoimmune diseases. pSGS mainly affects women over 40 years old, but can occur at any age. GSS is rare in children and is often secondary to other diseases. We report the case of a 12-year-old girl with primary Gougerot-Sjögren Syndrome Methods and results A 12-year-old girl from Batna (Algeria) presented to her pediatrician in 2016 for a rash with arthralgia. She had no relevant past medical history and is the child of a non-consanguineous marriage. Clinical examination found a well-nourished child with a purpuric rash on her lower limbs. She complained of arthralgia without clinical signs of arthritis. The child also complained of a sensation of a foreign body and burning in both eyes. The eye exam was normal except for a positive Schirmer's test. There were no signs of parotitis. Laboratory tests showed an erythrocyte sedimentation rate (ESR) of 84 mm the first h and a negative C-reactive protein (CRP). She was also Leukopenic. Serological testing resulted in a positive Anti SS-A and Anti SS-B. The abdominal pelvic ultrasound was normal. The patient was diagnosed with primary Gougerot-Sjögren Syndrome. Artificial tears and hydroxychloroquine were started as initial therapy. One year later we added an immunosuppressant (Methotrexate) due to the persistence of clinical signs. Conclusion Our case is particular in that primary Gougerot-Sjögren Syndrome is rare in children. Routine follow- up with this patient is important to determine whether it is indeed pGSS with pediatric onset or GSS secondary to another autoimmune disease.

Could omega-3 fatty acids preserve endothelial function in a rat model of rheumatoid arthritis?

Could omega-3 fatty acids preserve endothelial function in a rat model of rheumatoid arthritis? Background: Endothelial dysfunction is claimed to be the cause of increased risk for cardiovascular diseases in rheumatoid arthritis (RA) patients. Omega-3 fatty acids is a promising drug in this field; however, its exact effects on endothelial functions are not fully understood. Aim of the work: This study aimed to evaluate the effect of omega-3 fatty acids on endothelial reactivity and some markers of endothelial dysfunction [Vascular cell adhesion molecule-1 (VCAM-1), tumour necrosis factor alpha (TNF-α), Malondialdehyde (MDA) and NOS activity] in a rat model of rheumatoid arthritis. Material & methods: Thirty male rats were divided into 3 groups (control, untreated RA, and RA group with omega-3 supplementation). RA induction was done by intradermal injection of heat-killed Mycobacterium butyricum and was confirmed by clinical signs of arthritis on day 11. In the treated group, omega-3 was given daily via gastric gavage starting from day 11 until the end of the study. The study duration was 30 days for all groups starting from the day of induction. At the end of the study, rats were sacrificed, blood samples were collected for VACM-1, TNF-α, MDA and nitrite measurement and thoracic aortae were taken to test vascular reactivity. Results: All markers increased while vascular reactivity decreased significantly after RA induction. Omega-3 treatment significantly decreased all biochemical markers and restored normal vascular reactivity in the treated group.

Initial multi-target approach shows importance of improved caprine arthritis-encephalitis virus control program in Russia for hobbyist goat farms.

Background and Aim:Several reports described the detection of specific caprine arthritis-encephalitis virus (CAEV) antibodies in Russian goat populations, which indicates the circulation of CAEV in Russian goat farms. The aim of this study was to use a multi-target approach to testing with both serological tests and an in-house real-time (RT) molecular test to investigate the prevalence of CAEV in goats from three hobbyist farms in the Republic of Tatarstan, Russia.Materials and Methods:We applied a multi-target approach to testing with both enzyme-linked immunosorbent assay (ELISA) and an in-house RT polymerase chain reaction test to investigate the prevalence of CAEV in goats. Animals from the three hobbyist farms were used in this study. The animals from two farms (n=13 for F1 and n=8 for F2) had clinical signs of arthritis and mastitis. In the third farm (n=15 for F3), all goats were home-bred and had no contact with imported animals.Results:CAEV antibodies (ELISA targets TM env and gag genes) were detected in serum samples from two farms (F1 and F2), indicating seroprevalence of 87.50-92.31%. Specific CAEV antibodies were also detected in milk samples. CAEV proviral DNA was detected in 53.85-62.50%. The results from all tests performed in the third farm (F3) were negative, indicating that all tests were 100% specific.Conclusion:The results showed that CAEV is circulating and present in small hobbyist goat farms in Russia. Serological and molecular tests could be important for programs to control and eradicate CAEV in Russia for hobbyist goat farms.

СОВРЕМЕННЫЕ ВОЗМОЖНОСТИ УЛЬТРАЗВУКОВОЙ ДИАГНОСТИКИ ПСОРИАТИЧЕСКОГО АРТРИТА

Psoriatic arthritis is a heterogeneous disease characterized by the involvement of the axial skeleton and peripheral joints and / or entheses in the pathological process. The problems of diagnosing early psoriatic arthritis are not limited to the heterogeneity of clinical manifestations of the disease. Unlike rheumatoid arthritis, there are no biomarkers for the detection of early psoriatic arthritis, and therefore verification of the diagnosis depends on the identification of specific clinical signs. Finally, the absence of psoriasis in the presence of typical clinical signs of arthritis does not permit to confirm the diagnosis in the early stages. Considering numerous difficulties of clinical and laboratory diagnostics of psoriatic arthritis, great attention is paid to non-invasive instrumental methods for diagnosing the disease at early stages. The article provides up-to-date information on the potentials of ultrasound techniques in the diagnosis of early psoriatic arthritis.

Bone Erosions Detected by Ultrasound Are Prognostic for Clinical Arthritis Development in Patients With ACPA and Musculoskeletal Pain.

Anti-citrullinated protein antibodies (ACPA) often precede onset of rheumatoid arthritis (RA) by years, and there is an urgent clinical need for predictors of arthritis development among such at-risk patients. This study assesses the prognostic value of ultrasound for arthritis development among ACPA-positive patients with musculoskeletal pain. We prospectively followed 82 ACPA-positive patients without clinical signs of arthritis at baseline. Ultrasound at baseline assessed synovial hypertrophy, inflammatory activity by power Doppler, and erosions in small joints of hands and feet. We applied Cox regression analyses to examine associations with clinical arthritis development during follow-up (median, 69 months; range, 24–90 months). We also compared the ultrasound findings among the patients to a control group of 100 blood donors without musculoskeletal pain. Clinical arthritis developed in 39/82 patients (48%) after a median of 6 months (range, 1–71 months). One or more ultrasound erosions occurred in 13/82 patients (16%), with none in control subjects (p < 0.001). Clinical arthritis development was more common among patients with baseline ultrasound erosions than those without (77 vs. 42%, p = 0.032), and remained significant in a multivariable Cox regression analysis that included previously described prognostic factors (HR 3.9, 95% CI 1.6–9.4, p = 0.003). Ultrasound-detected tenosynovitis was more frequent among the patients and associated with clinical arthritis development in a univariable analysis (HR 2.5, 95% CI 1.1–5.7, p = 0.031), but did not remain statistically significant in multivariable analysis. Thus, bone erosions detected by ultrasound are independent predictors of clinical arthritis development in an ACPA-positive at-risk population.Trial Registration: Regional Ethics Committee in Linköping, Sweden, Dnr M220-09. Registered 16 December 2009, https://etikprovningsmyndigheten.se/.

The tellurium-based immunomodulator, AS101 ameliorates adjuvant-induced arthritis in rats.

Despite undeniable improvement in the management of rheumatoid arthritis (RA), the discovery of more effective, less toxic and, ideally, less immune suppressive drugs are much needed. In the current study, we set to explore the potential anti-rheumatic activity of the non-toxic, tellurium-based immunomodulator, AS101 in an experimental animal model of RA. The effect of AS101 was assessed on adjuvant-induced arthritis (AIA) rats. Clinical signs of arthritis were assessed. Histopathological examination was used to assess inflammation, synovial changes and tissue lesions. Very late antigen-4 (VLA-4)+ cellular infiltration was detected using immunohistochemical staining. Enzyme-linked immunosorbent assay (ELISA) was used to measure circulating anti-cyclic citrullinated-peptide autoantibody (ACPA) and real-time polymerase chain reaction (PCR) was used to measure the in-vitro effect of AS101 on interleukin (IL)-6 and IL-1β expression in activated primary human fibroblasts. Prophylactic treatment with intraperitoneal AS101 reduced clinical arthritis scores in AIA rats (P<0·01). AS101 abrogated the migration of active chronic inflammatory immune cells, particularly VLA-4+ cells, into joint cartilage and synovium, reduced the extent of joint damage and preserved joint architecture. Compared to phosphate-buffered saline (PBS)-treated AIA rats, histopathological inflammatory scores were significantly reduced (P<0·05). Furthermore, AS101 resulted in a marked reduction of circulating ACPA in comparison to PBS-treated rats (P<0·05). Importantly, AS101 significantly reduced mRNA levels of proinflammatory mediators such as IL-6 (P<0·05) and IL-1β (P<0·01) in activated primary human fibroblasts. Taken together, we report the first demonstration of the anti-rheumatic/inflammatory activity of AS101 in experimental RA model, thereby supporting an alternative early therapeutic intervention and identifying a promising agent for therapeutic intervention.

Investigation of anti-inflammatory effects of bee venom in experimentally induced adjuvant arthritis.

ObjectivesRheumatoid arthritis is a multisystemic inflammatory disease characterized by destruction of the joints. An effective treatment method of the disease has not been developed yet. The aim of the present study is to evaluate the effects of bee (Apis mellifera anatoliaca) venom (BV) on serum inflammatory parameters, serum antioxidant load and clinical parameters of experimentally induced adjuvant arthritis in rats.Material and methodsA total of 35 Wistar albino male rats were used. The animals were divided into 5 groups. First group animals served as negative controls. The second, third, fourth and fifth groups were used for experimental arthritis induction. Following clinical development of arthritis, the first group was subcutaneously administered 0.2 ml of physiological saline, and the second, third and fourth groups were treated subcutaneously with 2 µg/kg, 4 µg/kg and 20 µg/kg once a week three times. Physiological saline injected fifth group animals were used as a sham-treatment group. Clinical observations and evaluation of arthritis were made at the 15th day, and at the end of the experiment. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, paraoxonase, serum aryl esterase, high-sensitivity C reactive protein, interleukin 1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) were determined in cardiac blood samples taken at the end of the 29th day.ResultsFrom the data, total oxidant level (TOL) and oxidative stress index (OSI) were calculated. Significant improvements were observed in the clinical signs of arthritis and inflammatory markers such as in IL-1β, TNF-α, IL-6 and TOL and OSI in the 20.0 µg/kg BV-administered group. Bee venom administration did not cause any significant increase in ALT and AST values or signs of liver toxicity.ConclusionsBee venom treatment was effective in alleviation of symptoms of the experimental rat adjuvant arthritis by means of clinical observation and serum inflammatory markers.

It's never too soon to treat rheumatoid arthritis: finally, some supportive evidence.

For the past 30 years, early arthritis clinics have been promoted as a means of improving long-term outcomes for patients, 1 the logic being that early therapy minimises structural damage (which correlates with longer-term functional loss) induced by inflammation both locally and systemically. 2 Furthermore, there is the long-held belief in the, at least theoretical, window of opportunity whereby early intervention permits a qualitatively better outcome than the same intervention applied at a later date. 1 A recent focus on individuals at risk of rheumatoid arthritis has led to the realisation that multiple pathological mechanisms are taking place before clinical signs of arthritis—eg, objective evidence of subclinical inflammation on sensitive imaging, 3 and immunological abnormalities in T-cell subset numbers.

Identifying 'non-progressors' among patients with arthralgia.

Patients with clinically suspect arthralgia have articular symptoms such as pain and stiffness of the small joints without clinical signs of arthritis. Some of these patients progress and develop ‘true’ disease, but how can we differentiate evolving chronic disease from disease that will resolve?

Acellular Fish Skin Grafts and Pig Urinary Bladder Matrix Assessed in the Collagen-Induced Arthritis Mouse Model.

It is vital that cellular- and tissue-based products (CTPs) used for wound treatment do not provoke autoimmunity. In this study, the immunogenic response to extracts of 2 CTPs of piscine and porcine origin was assessed in the collagen-induced arthritis model. Male DBA/1J mice were divided into 4 groups, each composed of 7 to 9 animals. Each animal was injected with one of following to assess their immune responses: (1) bovine type II collagen (100 µg) in Freund's adjuvant, (2) extract of piscine skin (100 µg) in Freund's adjuvant, (3) extract of porcine urinary bladder matrix (100 µg) in Freund's adjuvant, or (4) Freund's adjuvant alone (control) at the beginning of the experiment and 3 weeks later. Clinical signs of arthritis were assessed from week 5 onwards, and anti-type II and anti-type I collagen antibody immunoglobulin G (IgG) serum levels were measured before injections and 8 weeks after exposure using enzyme-linked immunosorbent assays. Only the mice exposed to bovine type II collagen developed clinical arthritis accompanied by very high anti-type II collagen IgG serum levels. Anti-type II collagen IgG serum levels were also detected in the porcine group but were undetectable in the piscine skin and control groups after 8 weeks. There were no significant differences in anti-type I collagen IgG serum levels among the groups. The results showed that piscine skin did not provoke systemic autoimmunity against type II collagens in DBA/1J mice.

CCR6 controls autoimmune but not innate immunity-driven experimental arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease, characterized by synovial infiltration of various inflammatory cells. Chemokines are involved in controlling the recruitment of different cell types into the synovial membrane. The role of CCR6 in the development of arthritis so far remains unclear. In this study, we investigated the role of CCR6 in the pathogenesis of arthritis using three different murine arthritis models. Compared to WT animals, CCR6 −/− mice developed less clinical signs of arthritis in the collagen‐induced arthritis model but not in the K/BxN serum transfer arthritis model and in the human tumour necrosis factor transgenic arthritis model, suggesting a defect in adaptive effector functions but intact innate effector functions in the development of arthritis in CCR6 −/− animals. In line with this, anti‐collagen antibody levels were significantly reduced in CCR6 −/− mice compared with WT mice. Moreover, we demonstrate enhanced osteoclastogenesis in vitro in CCR6 −/− mice compared with WT mice. However, we did not detect differences in bone mass under steady state conditions in vivo between WT and CCR6‐deficient mice. These data suggest that CCR6 is crucially involved in adaptive but not in innate immunity‐driven arthritis. CCR6 or its chemokine ligand CCL20 might represent a possible new target for the treatment of RA.

Power Doppler Ultrasound Evaluation of Peripheral Joint, Entheses, Tendon, and Bursa Abnormalities in Psoriatic Patients: A Clinical Study.

To evaluate the prevalence rates of peripheral joint, enthesis, tendon, and bursa abnormalities by power Doppler (PD) ultrasonic examination in patients with psoriatic arthritis (PsA), psoriatic patients without clinical signs of arthritis (non-PsA psoriasis group), and healthy individuals, to detect subclinical PsA. A total of 253 healthy volunteers, 242 non-PsA psoriatic patients, and 86 patients with PsA were assessed by 2-dimensional and power Doppler (PD) ultrasound. Peripheral joint, enthesis, tendon, and bursa abnormalities were observed, characterizing abnormal PD. The affected patients and sites with abnormalities in various ages were compared among groups; PD signal grades for the abnormalities were also compared. In the PsA group, significantly higher percentages of sites showing joint effusion/synovitis, enthesitis, and tenosynovitis in all age groups, and markedly higher rates of sites with bursitis were found in young and middle age groups, compared with the non-PsA and control groups (all p < 0.01). Meanwhile, the non-PsA group showed significantly higher rates of joint effusion/synovitis and enthesitis sites, and elevated PD signal grades of synovitis, enthesitis, and tenosynovitis in comparison with the control group, both in young and middle age groups (all p < 0.01). Patients with PsA have high percentages and PD signal grades of peripheral joint, tendon, enthesis, and bursa involvement. Young and middle-aged non-PsA patients have high synovitis and enthesitis percentages, and elevated PD signal grades of synovitis, enthesitis, and tenosynovitis.

Cartilage damage and bone erosion are more prominent determinants of functional impairment in longstanding experimental arthritis than synovial inflammation.

ABSTRACTChronic inflammation of articular joints causing bone and cartilage destruction consequently leads to functional impairment or loss of mobility in affected joints from individuals affected by rheumatoid arthritis (RA). Even successful treatment with complete resolution of synovial inflammatory processes does not lead to full reversal of joint functionality, pointing to the crucial contribution of irreversibly damaged structural components, such as bone and cartilage, to restricted joint mobility. In this context, we investigated the impact of the distinct components, including synovial inflammation, bone erosion or cartilage damage, as well as the effect of blocking tumor necrosis factor (TNF) on functional impairment in human-TNF transgenic (hTNFtg) mice, a chronic inflammatory erosive animal model of RA. We determined CatWalk-assisted gait profiles as objective quantitative measurements of functional impairment. We first determined body-weight-independent gait parameters, including maximum intensity, print length, print width and print area in wild-type mice. We observed early changes in those gait parameters in hTNFtg mice at week 5 – the first clinical signs of arthritis. Moreover, we found further gait changes during chronic disease development, indicating progressive functional impairment in hTNFtg mice. By investigating the association of gait parameters with inflammation-mediated joint pathologies at different time points of the disease course, we found a relationship between gait parameters and the extent of cartilage damage and bone erosions, but not with the extent of synovitis in this chronic model. Next, we observed a significant improvement of functional impairment upon blocking TNF, even at progressed stages of disease. However, blocking TNF did not restore full functionality owing to remaining subclinical inflammation and structural microdamage. In conclusion, CatWalk gait analysis provides a useful tool for quantitative assessment of functional impairment in inflammatory destructive arthritis. Our findings indicate that cartilage damage and bone erosion, but not synovial inflammation, are the most important determinants for progressive functional impairment in this chronic erosive arthritis model.

Incidence of Rheumatoid Arthritis Onset in Patients with Arthralgia and Anti- Citrullinated Peptide Antibody Positivity: Pilotstudy on Effectiveness of Hydroxychloroquin Treatment

Objective: A relevant subset of patients with ACPA-positive arthralgia will develop arthritis within one year. Hence, an ACPA-positive patient cohort with arthralgia was treated with hydroxychloroquine (HCQ) and was followed-up in terms of RA-manifestation and arthralgia complaints. Methods: Since 2009 all patients presenting with arthralgia showing positive ACPA without clinical signs of arthritis were investigated retrospectively at a large outpatient facility in Lower Saxony, Germany. Body-weight adapted hydroxychloroquine-therapy was started at outset and assessments comprise a detailed documentation of arthralgias and rheumatological status (DAS28). The RA-onset rate within 12 months after enrollment is regarded as the primary outcome; secondary outcomes are the clinical course and humoral inflammatory activity. Results: Within 12 months one of 24 patients developed RA (5%), one further patient within 24 months (10%). Regarding the secondary outcomes the number of arthralgic sites decreased highly significant, the DAS28-score and the rheumatoid factor improved after both 12 and 24 months follow-up (p<0.05). Furthermore, the mean steroid dosage could be tapered from 5 mg to 1 mg, and to 0 mg, respectively. In contrast to the present pilot study, conventional treatment (without DMARDs) in seropositive arthralgia patients yielded about 6-fold and 3-fold higher rates of RA-incidence within 12 months and 24 months, respectively. Conclusion: Since these results are promising, randomized controlled studies are needed to evaluate the effects of HCQ on RA-prevention in seropositive arthralgia patients and to define its role regarding the clinical course of seropositive arthralgia within a possible “window of opportunity”.

The differential impact of TH+ neuronal cell therapy in models of experimental arthritis

Background A long-lasting anti-inflammatory effect of generated catecholamine-producing tyrosine hydroxylase-positive (TH+) cells was shown in collagen type II-induced arthritis (CIA). Here, we investigated the importance of TH+ cells in another model of rheumatoid arthritis. Methods TH+ cells were generated from murine bone -derived mesenchymal stem cells by specific catecholaminergic factors. Antigen-induced arthritis (AIA) was induced in C57BL/6J mice and one group was treated by TH+ cell-transfer. The appearance of natural TH+ cells, clinical signs of arthritis, pain and immune parameters were observed during AIA. Results In contrast to CIA, here, only a small number of natural occurring TH+ cells were detectable in spleen in the acute phase of arthritis. After TH+ cell transfer, joint swelling was only moderately reduced in acute AIA, but mice showed significantly less guarding and less reduction of withdrawal threshold for mechanical stimulation on the inflamed hind limb. These effects were attenuated towards later phases of the disease. Furthermore both, in TH+ treated mice cytokines in knee joint extracts and immunoglobulins and cytokines in sera were not reduced in acute AIA. Moreover, significantly enhanced values of IL-17, IgG1 and IgG3 were detected. Conclusion In acute AIA, TH+ cells have rather an anti-nociceptive than an anti-inflammatory effect. The increase of inflammatory parameters corresponds to the effect of the sympathetic system on the acute phase of the AIA.

MSC signature associated with therapeutical effect in osteoarticular diseases

Adipose derived mesenchymal stromal cells (ASC) are adult stem cells exhibiting functional properties that have open the way for cell-based clinical therapies. Primarily, their capacity of multilineage differentiation has been explored in a number of strategies for skeletal tissue regeneration. More recently, MSCs have been reported to exhibit immunosuppressive as well as healing capacities, to improve angiogenesis and prevent apoptosis or fibrosis through the secretion of paracrine mediators including HGF, IL1RA, IL-6, PDL-1, TSG6, TGF-β1 and PGE2. In normal homeostasis, IL1Ra counterbalances the effect of IL1α and IL1β as a competitive receptor antagonist. MSC immunosuppressive effect is highly variable according to cell population heterogeneity. We compared gene expression of huMSC with high potential to inhibit T cells response with low efficient MSC through gene array. We identified a signature associated with high immunoregulatory function. High expression of ITPR1, NR4A3, ABCA1, LIPG ADAMTS4 as well as expression of CD109, ITGA10, IL6R was associated with increased T cell suppressive effect in vitro. We performed preclinical models of osteoarthritis, and showed that a local injection of ASC showed a reduction of synovitis, reduction of osteophytes, joint stabilization, reducing the score of cartilage lesions. We then investigated the role of iNOS, IL6 and IL1RA respectively using the inflammatory collagen-induced arthritis model. In contrast to wt MSCs, which reduced both the incidence and clinical signs of arthritis, IL1RA–/– MSCs were not able to prevent arthritis progression and even worsened the arthritic symptoms. μCT analysis showed a protection against bone erosion by wt MSCs but not in IL1RA–/–MSCs-treated mice. We found a decrease of the number of CD4 + IFN-γ+ Th1 and CD4 + IL-17+ Th17 lymphocytes in the spleens of wt MSCs-treated mice as compared to IL1RA–/– MSCs-treated mice. This work was completed by toxicology data showing the excellent tolerance of the local injection of ADSC and biodistribution showing the persistence of cells after 6 months in murine models. We conducted a open-label phase 1 trial including 18 patients with severe osteoarthritis of the knee in failure of conventional therapies (62.5% were KL IV) at two sites, Montpellier and Wurzburg. Mean age was 61 years, with a 10 years history of knee OA. The patient received a single injection of autologous ASC 15 days after lipoaspiration (2.106, 107 or 5.107) through intra-articular injection. The primary outcome measure of effectiveness was patient-reported WOMAC pain subscores by VAS in the affected knee at week 12. Secondary outcome measures included Outcome Measures in Rheumatology Clinical Trials and Osteoarthritis Research Society International (OMERACT OARSI) responses. We observed a decrease of the VAS Pain (73 ± 11 mm day 0 to 32 ± 23 month 3), and of WOMAC (50 ± 18 to 25 ± 7 month 3). This study confirms the feasibility and safety of local injection of autologous cells from adipose tissue and suggested that the most effective dose was 107 autologous cells. Moreover, a molecular signature associated with increase immunosuppressive potential is described.

Blockage of lysophosphatidic acid reverses arthritis-induced hypersensitivity and Cavα2δ1 and P2X3 expression in dorsal root ganglia

Abstract Aims The lysophosphatidic acid (LPA) is an important mediator involved in neuropathic and bone cancer pain models. We are investigating if LPA is involved in arthritis-induced pain in the collagen antibody-induced arthritis (CAIA) mice model. Methods Arthritis was induced in male CBA mice by injection of 1.5 mg collagen type II antibody cocktail. Mechanical and thermal sensitivity and the degree of arthritis were assessed with von Frey filaments, Hargreaves box (heat), acetone test (cold) and visual scoring, respectively. LPA antibody and control IgG (10mg/kg), or saline was injected s.c. twice a week from day 12 through day 47. qPCR and immunohistochemical studies were undertaken in dorsal root ganglia (DRGs) to explore the expression of pain-related ion channels. Results Administration of LPA antibody treatment reversed CAIA-induced mechanical and thermal hypersensitivity (p &lt; 0.05) while had no effect on the early clinical signs of arthritis (p &gt; 0.05). mRNA levels for the LPA synthesizing enzyme autotaxin were elevated in the CAIA group. On day 48, expression of the voltage-gated calcium channel Cavα2δ1 and the ATP-gated P2X3 receptor were significantly increased in the CAIA DRGs, which were completely prevented by LPA antibody treatment. Of note, based on in vitro experiment, LPA stimulation upregulated Cavα2δ1 and P2X3 expression in primary adult mouse DRG cultures. Conclusions Blocking the action of systemic LPA reverses arthritis-induced hypersensitivity, potentially through regulation of Cavα2δ1 and P2X3 expression in peripheral neurons. Thus, our data point to that LPA may serve as a target for providing pain relief in arthritis.