Exploring the involvement of NLRP3 and Il-1β in Osteoarthritis of the Hand: Results from a Pilot Study

Hand osteoarthritis (HOA) includes different subsets; a particular and uncommon form is erosive HOA (EHOA). Interleukin- (IL-) 1 plays a crucial role in the pathogenesis of osteoarthritis (OA); it is synthesized as an inactive precursor which requires the intervention of a cytosolic multiprotein complex, named inflammasome, for its activation. The aim of this study was to investigate the involvement of IL-1 and the NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome in patients with EHOA and nonerosive HOA (NEHOA) compared to healthy controls. In particular, we evaluated the gene expression of IL-1 and NLRP3, the serum levels of IL-1, IL-6, IL-17, and tumor necrosis factor- (TNF-) , and the protein levels of IL-1 and NLRP3. We also assessed the relationships between IL-1 and NLRP3 and clinical, laboratory, and radiological findings. Fifty-four patients with HOA (25 EHOA and 29 NEHOA) and 20 healthy subjects were included in the study. Peripheral blood mononuclear cell (PBMC) gene and protein expressions of IL-1 and NLRP3 were quantified by quantitative real-time PCR and western blot. IL-1, IL-6, IL-17, and TNF- serum levels were determined by ELISA. IL-1 gene expression was significantly reduced (p=0.0208) in EHOA compared to healthy controls. NLRP3 protein levels were significantly increased in the NEHOA group versus the control (p=0.0063) and EHOA groups (p=0.0038). IL-1 serum levels were not significantly different across the groups; IL-6, IL-17, and TNF- were not detectable in any sample. IL-1 concentrations were negatively correlated with the Kellgren-Lawrence score in the whole population (r=-0.446; p=0.0008) and in NEHOA (r=-0.608; p=0.004), while IL-1 gene expression was positively correlated with the number of joint swellings in the EHOA group (r=0.512; p=0.011). Taken together, our results, showing poorly detectable IL-1 concentrations and minimal inflammasome activity in the PBMCs of HOA patients, suggest a low grade of systemic inflammation in HOA. This evidence does not preclude a possible involvement of these factors at the local level

Prescription-grade crystalline glucosamine sulfate as an add-on therapy to conventional treatments in erosive osteoarthritis of the hand: results from a 6-month observational retrospective study.

peer reviewed[en] OBJECTIVE: To evaluate the efficacy of prescription-grade Crystalline Glucosamine Sulfate (pCGS) as an add-on treatment to conventional therapy, compared to usual therapy alone, in patients with erosive osteoarthritis of the hand (EHOA). METHODS: This 6-month retrospective case-control study included patients with concomitant knee osteoarthritis and symptomatic EHOA. Participants were stratified into two groups based on whether or not pCGS (1500 mg/day) was added to the conventional therapy (education and training in ergonomic principles, exercise and use on-demand of symptomatic drugs) for hand osteoarthritis. Patients were evaluated at baseline, after 3 and 6 months. Primary outcomes were the change from baseline to month 6 in Visual Analogue Scale (VAS) hand pain and in Functional Index for Hand Osteoarthritis (FIHOA) score. A set of secondary parameters was also evaluated. RESULTS: 123 patients were included as follows: 67 treated with pCGS in addition to conventional therapy (pCGS Group) and 56 with conventional therapy alone (Control Group). After 6 months a significant difference in VAS and in FIHOA score (p < 0.01 and p < 0.001, respectively) was observed in favor of pCGS Group. Similar results were found for morning stiffness duration (p < 0.05), health assessment questionnaire (p < 0.01) and physical and mental component score of 36-item short form (p < 0.05 and p < 0.001, respectively). A significant reduction of symptomatic drug consumption at 3 and 6 months was reported in the pCGS Group (p < 0.001). No serious adverse event was recorded in both groups. CONCLUSIONS: Despite all the limitations inherent to an observational study, our results suggest the potential effectiveness of pCGS, when used in combination with conventional therapy in EHOA. Further randomized placebo-controlled trials are needed to confirm these positive findings. TRIAL REGISTRATION: ClinicalTrials.gov, http://www. CLINICALTRIALS: gov , date of registration: February 2, 2022, NCT05237596. The present trial was retrospectively registered

Could oxidative stress regulate the expression of microRNA-146a and microRNA-34a in human osteoarthritic chondrocyte cultures?

Oxidative stress and the overproduction of reactive oxygen species (ROS) play an important role in the pathogenesis of osteoarthritis (OA). Accumulating evidence has demonstrated the involvement of microRNAs (miRNAs) dysregulation in disease development and progression. In this study, we evaluated the effect of oxidative stress on miR-146a and miR-34a expression levels in human OA chondrocytes cultures stimulated by H2O2. Mitochondrial ROS production and cell apoptosis were detected by flow cytometry. The antioxidant enzymes SOD-2, CAT, GPx, the transcriptional factor NRF2 and the selected miRNAs were analyzed by qRT-PCR. The H2O2-induced oxidative stress was confirmed by a significant increase in superoxide anion production and of the apoptotic ratio. Furthermore, H2O2significantly up-regulated the expression levels of SOD-2, CAT, GPx and NRF2, and modulated miR-146a and miR-34a gene expression. The same analyses were carried out after pre-treatment with taurine, a known antioxidant substance, which, in our experience, counteracted the H2O2-induced effect. In conclusion, the induction of oxidative stress affected cell apoptosis and the expression of the enzymes involved in the oxidant/antioxidant balance. Moreover, we demonstrated for the first time the modification of miR-146a and miR-34a in OA chondrocytes subjected to H2O2stimulus and we confirmed the antioxidant effect of taurine

Long-term complete response in a patient with liver metastases from breast cancer treated with metronomic chemotherapy

Background. Preclinical studies have shown that several chemotherapeutic agents at low doses may affect the vascular system. Here we report the case of a patient with long-term cancer control by metronomic chemotherapy. Case presentation. A 62-year-old woman with breast cancer underwent a left mastectomy in July 2007. For a liver metastasis she was given first-line chemotherapy with doxorubicin plus paclitaxel every 21 days. A CT scan after the sixth cycle showed a partial response. It was decided to stop the treatment with doxorubicin and paclitaxel, and start metronomic therapy with cyclophosphamide 50 mg daily orally and methotrexate 2.5 mg twice daily, 2 days a week. After 6 months of this maintenance treatment, CT scan showed a complete response. We examined the expression of vascular endothelial growth factor receptor 2 (VEGFR2) in histological sections of the primary tumor of our patient, finding evidence of overexpression of the receptor. The metronomic treatment is still ongoing, and after 60 months the patient maintains a complete response. Conclusion. This clinical case highlights how suitable metronomic chemotherapy can be used as maintenance therapy, allowing long-term treatment with no significant toxicity. This case suggests that the level of VEGFR2 is predictive of best response to antiangiogenic therapy

Identification of MOR-Positive B Cell as Possible Innovative Biomarker (Mu Lympho-Marker) for Chronic Pain Diagnosis in Patients with Fibromyalgia and Osteoarthritis Diseases

Fibromyalgia (FM) diagnosis follows the American College of Rheumatology (ACR) criteria, based on clinical evaluation and written questionnaires without any objective diagnostic tool. The lack of specific biomarkers is a tragic aspect for FM and chronic pain diseases in general. Interestingly, the endogenous opioid system is close to the immune one because of the expression of opioid receptors on lymphocytes membrane. Here we analyzed the role of the Mu opioid receptor on B lymphocytes as a specific biomarker for FM and osteoarthritis (OA) patients. We enrolled three groups of females: FM patients, OA patients (chronic pain control group) and healthy subjects (pain-free negative control group). We collected blood samples to apply immunophenotyping analysis. Written tests were administrated for psychological analysis. Data were statistically analyzed. Final results showed that the percentage of Mu-positive B cells were statistically lower in FM and OA patients than in pain-free subjects. A low expression of Mu-positive B cell was not associated with the psychological characteristics investigated. In conclusion, here we propose the percentage of Mu-positive B cells as a biological marker for an objective diagnosis of chronic pain suffering patients, also contributing to the legitimacy of FM as a truly painful disease

An observational study on chronic pain biomarkers in fibromyalgia and osteoarthritis patients:.which role for mu opioid receptor’s expression on NK cells.

The evaluation of chronic pain is challenging because of the lack of specific biomarkers. We identified the Mu opioid receptor-positive (Mu+) B cell percentage of expression, named Mu-Lympho-Marker (MLM), as a candidate marker for chronic pain in fibromyalgia (FM) and osteoarthritis (OA) patients. Here, we investigate the role of MLM on natural killer (NK) cells in the same patients. Twenty-nine FM and twelve OA patients were analyzed, and twenty-three pain-free subjects were considered as the control group. Blood samples were collected to perform immunophenotyping and Western blot analysis. Biological and clinical data were statistically analyzed. The final results showed that the percentage of NK cells expressing Mu was statistically lower in FM and OA patients than in pain-free subjects, as already demonstrated for B cells. A Western blot analysis was performed in order to detect NK cells' functional status. Moreover, the correlation analysis of MLM expression with pharmacological therapy did not show any significant results. In conclusion, here, we confirm the role of MLM as a suitable marker for chronic pain and underline NK cells as a new possible immune cell type involved in the "Mu opioid receptor reserve theory"

Research and Development for Near Detector Systems Towards Long Term Evolution of Ultra-precise Long-baseline Neutrino Experiments

With the discovery of non-zero value of $\theta_{13}$ mixing angle, the next generation of long-baseline neutrino (LBN) experiments offers the possibility of obtaining statistically significant samples of muon and electron neutrinos and anti-neutrinos with large oscillation effects. In this document we intend to highlight the importance of Near Detector facilities in LBN experiments to both constrain the systematic uncertainties affecting oscillation analyses but also to perform, thanks to their close location, measurements of broad benefit for LBN physics goals. A strong European contribution to these efforts is possible

IL COINVOLGIMENTO DELL'IL-1β E DELL'INFLAMMASOMA NLRP3 NELL'OSTEOARTROSI EROSIVA E NODALE DELLA MANO

RIASSUNTO Background: L’osteoartrosi (OA) della mano, tipica espressione della malattia nella sua forma primaria, racchiude un eterogeneo gruppo di “affezioni", tra cui l'Osteoartrosi Erosiva (OAE), definita dalla presenza di peculiari caratteristiche radiologiche, quali le erosioni centrali delle articolazioni interfalangee prossimali e distali e da una spiccata componente flogistica. L'Interleuchina (IL)-1β svolge un ruolo chiave nella patogenesi dell'OA, stimolando il rilascio da parte dei condrociti dei principali enzimi responsabili del catabolismo cartilagineo e inibendo la produzione dei componenti della matrice extracellulare. L'IL-1β è sintetizzata come precursore inattivo e la sua attivazione avviene ad opera del complesso "Inflammosoma", la cui variante più conosciuta è rappresentata dall'Inflammasoma NLRP3. Negli ultimi anni, diversi studi si sono focalizzati sul ruolo dell'NLRP3 nelle malattie autoinfiammatorie e autoimmuni, mentre pochi hanno indagato il suo possibile coinvolgimento nella patogenesi dell'OA. Scopo del lavoro: Scopo del nostro studio è stato quello di valutare il possibile ruolo dell'IL-1β e del complesso dell'Inflammosoma NLRP3 in un gruppo di pazienti affetti da OAE e in un gruppo di pazienti con OA nodale (OAN) della mano, in comparazione con un gruppo di soggetti sani. In particolare, abbiamo analizzato l'espressione genica dell'IL-1β e dell'NLRP3, i livelli proteici dell'NLRP3 e le concentrazioni sieriche dell'IL-1β e di altre citochine pro-infiammatorie (IL-6, IL-17 e TNF-α) dimostrate implicate nella patogenesi dell'OA. Infine, abbiamo valutato le possibili correlazioni tra l'IL-1β e l'NLRP3 e parametri clinici, di laboratorio e radiologici dei pazienti in studio. Materiali e metodi: Nel nostro studio venivano inclusi 54 pazienti affetti da OA della mano, secondo i criteri ACR, di cui 25 con OAE e 29 con OAN e un gruppo di 20 soggetti sani comparabili per sesso ed età. L'espressione genica dell'IL-1β e dell'NLRP3 e i livelli proteici dell'NLRP3 venivano valutati nelle cellule mononucleate, derivate dal sangue periferico dei pazienti. L'espressione genica veniva quantificata tramite real-time PCR quantitativa, mentre il Western Blot permetteva la determinazione dei livelli proteici dell'NLRP3. La concentrazione sierica dell'IL-1β, IL-6, IL-17 e TNF-α veniva valutata tramite test ELISA. Il test di Mann-Whitney e il test di Fisher venivano utilizzati per analizzare le differenze tra variabili continue; le correlazioni tra le diverse variabili cliniche e laboratoristiche sono state effettuate tramite il test di Pearson; valori di p<0,05 sono stati considerati significativi. Risultati: I livelli serici di IL-1β, non mostravano differenze significative tra i tre gruppi in studio, mentre l’espressione genica dell’IL-1β risultava significativamente più bassa (p=0,0263) nei pazienti con OAE, rispetto al gruppo di controllo. Nella valutazione dell'espressione genica dell'NLRP3, non si apprezzavano differenze significative tra i tre gruppi, mentre i livelli proteici dell'NLRP3 risultavano significativamente più elevati (p=0,0063) nei pazienti con OAN, rispetto ai controlli. Inoltre, i livelli proteici dell'NLRP3 risultavano significativamente maggiori (p=0,0038) nei pazienti affetti da OAN, rispetto ai pazienti con OAE. I livelli sierici dell'IL-6, IL-17A e TNF-α risultavano al di sotto della soglia minima rilevabile dal test ELISA in tutti i campioni analizzati. Lo studio delle correlazioni evidenziava una correlazione negativa, statisticamente significativa, tra le concentrazioni seriche di IL-1β e il grado radiologico, secondo Kellgren e Lawrence, nell'intera popolazione studiata (r=-0,446;p=0,0008) e nel gruppo di pazienti con OAN (r=-0,608;p=0,004). Infine, l'espressione genica dell'IL-1β correlava positivamente, in maniera statisticamente significativa, con il numero di articolazioni tumefatte della mano nel gruppo di pazienti con OAE (r=0,512;p=0,011). Conclusioni: Il nostro studio, mostrando bassi valori sia dei livelli sierici, sia dell'espressione genica dell’IL-1β in pazienti affetti da OAE e OAN, supporta l’ipotesi di un meccanismo indipendente dall’IL-1 nello sviluppo del processo di degradazione cartilaginea e dell’infiammazione nell’OA della mano. Inoltre, la bassa espressione proteica dell’NLRP3 riscontrata nei soggetti con OAE, rispetto ai pazienti con OAN e ai controlli, ci induce ad ipotizzare un ruolo, non così centrale, dell’NLRP3 nella patogenesi dell’OAE. ABSTRACT Background: Osteoarthritis of the hand (HOA) is a relatively common form of osteoarthritis (OA). Though less common than non-erosive HOA (NEHOA), erosive HOA (EHOA) is characterized by preminent inflammatory features and greater disability. There is much speculation as to whether inflammatory pathways play a role in the pathophysiology of EHOA. IL-1β plays a crucial role in the pathogenesis of OA leading to the release of cartilage degrading enzymes such as metalloproteinases and aggrecanases from chondrocytes and inhibiting the production of extracellular matrix. IL-1β is synthesized as an inactive precursor which requires the intervention of a large cytosolic multi-protein complexes, named inflammasomes for its activation. Objective: The aim of this study was to investigate the role of IL-1β and the NLRP3 inflammasome in patients with EHOA and NEHOA in comparison to healthy controls. In particular, we evaluated the gene expression of NLRP3 and IL-1β, the serum levels of IL-1β, IL-6, IL-17 and TNF-α and the protein levels of NLRP3. We also assessed the relationships between IL-1β and NLRP3 and other clinical, laboratory and radiological parameters studied in EHOA and NEHOA patients. Patients and Methods: fifty-four patients with HOA, 25 with EHOA and 29 with NEHOA, and a control group of 20 healthy subjects were included in the current study. Blood samples from each subject were collected. Peripheral blood mononuclear cell (PBMC) expression of NLRP3 and IL-1β mRNA was quantified across the study subgroups by quantitative real-time PCR and NLRP3 PBMCs protein levels by Western Blot. IL-1β, IL-6, IL-17 and Tumor Necrosis Factor (TNF)-α serum levels were determined by ELISA. Results: Significantly lower IL1-β gene expression was observed in the EHOA patient group compared to healthy controls (p=0.0263). NLRP3 protein levels were significantly elevated in the NEHOA group versus controls (p=0.0063) and EHOA groups (p=0.0038). IL-1β concentration was not significantly different across the groups; IL-6, IL-17 and TNF-α were not detectable in any sample. IL-1β concentration was negatively correlated with Kellgren-Lawrence score in the whole population (r= -0.446; p=0.0008) and in the NEHOA (r= -0.608; p=0.004), while IL-1β gene expression was positively correlated with the number of hand joint swellings in the EHOA group (r=0.512; p=0.011). Conclusions: Low IL-1β serum concentration and gene expression in EHOA patients in particular, suggest that the mechanism involved in cartilage degradation and inflammation is independent of IL-1 pathways. Lower NLRP3 protein expression in EHOA patients, compared to NEHOA and controls, suggest NLRP3 is also not critical in the pathogenesis of EHOA

Spa therapy: can be a valid option for treating knee osteoarthritis?

Osteoarthritis (OA) continues to be one of the leading causes of 'years lived with disability' worldwide. Symptomatic knee OA is highly prevalent among people aged 50 years and over and is destined to become an ever more important healthcare problem. Current management of knee OA includes non-pharmacological and pharmacological treatments. Spa therapy is one of the most commonly used non-pharmacological approaches for OA in many European countries, as well as in Japan and Israel. Despite its long history and popularity, spa treatment is still the subject of debate and its role in modern medicine continues to be unclear. The objective of this review is to summarize the currently available information on clinical effects and mechanisms of action of spa therapy in knee OA. Various randomized controlled clinical trials (RCTs) were conducted to assess the efficacy and tolerability of balneotherapy and mud-pack therapy in patients with knee OA. Data from these clinical trials support a beneficial effect of spa therapy on pain, function and quality of life in knee OA that lasts over time, until 6-9 months after the treatment. The mechanisms by which immersion in mineral or thermal water or the application of mud alleviate suffering in OA are not fully understood. The net benefit is probably the result of a combination of factors, among which the mechanical, thermal and chemical effects are most prominent. In conclusion, spa therapy seems to have a role in the treatment of knee OA. Additional RCTs and further studies of mechanisms of action with high methodological quality are necessary to prove the effects of spa therapy