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Beer consumption has been identified as a risk factor for osteoarthritis (OA), a rheumatic disease characterised by cartilage degradation, joint inflammation, and eventual joint failure. One of the main isoflavonoids in beer is formononetin (FNT), an estrogenic compound also found in multiple plants and herbs. In this study, we aimed to investigate the effect of FNT on chondrocyte viability, inflammation, and metabolism. Cells were treated with FNT with or without IL-1β for 48 h and during 7 days of differentiation. Cell viability was determined via MTT assay. Nitrite accumulation was determined by Griess reaction. The expression of genes involved in inflammation and metabolism was determined by RT-PCR. The results revealed that a low concentration of FNT had no deleterious effect on cell viability and decreased the expression of inflammation-related genes. However, our results suggest that FNT overexposure negatively impacts on chondrocytes by promoting catabolic responses. Finally, these effects were not mediated by estrogen receptors (ERs) or aryl hydrocarbon receptor (AhR). In conclusion, factors that favour FNT accumulation, such as long exposure times or metabolic disorders, can promote chondrocyte catabolism. These data may partially explain why beer consumption increases the risk of OA.

Osteoarthritis, a major cause of disability in developed countries does not have effective treatment. Activation of TLR4 and innate immune response factors contribute to osteoarthritis progressive cartilage degradation. There are no clinically available TLR4 inhibitors. Interestingly, the antidepressant amitriptyline could block this receptor. Thus, we evaluated amitriptyline anti-TLR4 effects on human osteoarthritis chondrocytes in order to repurpose it as an inhibitor of innate immune response in joint inflammatory pathologies.Using in silico docking analysis, RT-PCR, siRNA, elisa, proteomics and clinical data mining of drug consumption, we explored the clinical relevance of amitriptyline blockade of TLR4-mediated innate immune responses in human osteoarthritis chondrocytes.Amitriptyline bound TLR4 but not IL-1 receptor. Interestingly, amitriptyline binding to TLR4 inhibited TLR4- and IL-1 receptor-mediated innate immune responses in human osteoarthritis chondrocytes, synoviocytes and osteoblasts cells. Amitriptyline reduced basal innate immune responses and promoted anabolic effects in human osteoarthritis chondrocytes. Supporting its anti-innate immune response effects, amitriptyline down-regulated basal and induced expression of NLRP3, an inflammasome member from IL-1 receptor signalling linked to osteoarthritis and gout pathologies. Accordingly, mining of dissociated and aggregated drug consumption data from 107,172 elderly patients (65 years) revealed that amitriptyline consumption was significantly associated with lower colchicine consumption associated with inflammatory gout flare treatment.Amitriptyline blocks TLR4-, IL-1 receptor and NLRP3-dependent innate immune responses. This together with clinical data amitriptyline could be repurposed for systemic or local innate immune response management in diverse joint inflammatory pathologies.

This study investigates the biological response of tantalum and titanium materials with Laser Induced Periodic Surface Structures (LIPSS) produced by a femtosecond laser at a wavelength of 1030 nm. Periodic surface nanostructures were formed by laser irradiation using a laser energy slightly above the damage threshold fluence for titanium and tantalum; pulses with a 500 fs pulse duration at a high repetition rate were applied to smooth polished tantalum and titanium surfaces with an original roughness value of 3.9 ± 0.2 nm and 3.1 ± 0.2 nm, respectively. We have demonstrated that LIPSS are an efficient option to increase osteoblastic differentiation of hBMSCs in vitro on titanium and tantalum surfaces. LIPSS modification created a statistically significant increase in cell metabolism; the best values were observed using an MTT assay. Similar improvements in inflammatory response to the material (IL-6 and TNF-alfa values) were also observed. Results also show that collagen is produced in greater quantities and cells differentiate to osteoblasts easily. These differences are seen from the beginning until the end point of the in-vitro trial over a 20 day duration.

Background:Osteoarthritis (OA) incidence has skyrocketed in the last decade and yet a definitive treatment has still to be found. This worldwide disease is depriving our society from their life quality and has become a grave economic burden. Research on anti-inflammatory tools has been done on traditional Asian medicine. Boswellic acid is a plant-derived molecule from the Boswellia species that has shown to prevent cartilage loss in an OA mouse model[1]. However, the specific mechanism of action is still unclear. The activation of innate immune receptors, such Toll-like receptor 4 (TLR4) has been involved in chondrocyte-mediated inflammatory responses and OA development. Although, boswellic acid has shown an inhibitory effect on TLR4-mediated inflammatory responses little is known about its role on TLR4-mediated chondrocyte inflammatory and catabolic responses.Objectives:Determine the ability of beta boswellic acid (BBA) to block TLR4-mediated innate immune responses in chondrocytes and synoviocytes.Methods:In silicoThe binding affinity of beta boswellic acid (BBA) to TLR4 complex signalling was determined by optimized docking algorithm in the BIO-HPC Research Group facilities.In vitroCellular proteome and secretome profiling (LC-MALDI/TOFF) was used to study inflammatory pathways induced by the agonist of TLR4 (LPS [100ng/ml]) and IL1R (IL1β [0.1ng/ml]). The effect of BBA on TLR4-mediated innate immune responses was determined by RT-PCR, Western Blot and ELISA in primary human OA chondrocytes (hOC), murine ATDC5 chondrocytes, human synoviocytes (SW982) and primary human osteoblasts (hOB). Cell viability was tested using the methyl-thiazolyl-tetrazolium (MTT) reagent. Nitric oxide production in cell culture media was assessed by Griess reaction. Green Malachite Assay was used to semi-quantify the whole phosphoproteome.EthicsThis study was approved by the CEIC (CAEIG 2014/310).Results:Cellular proteome and secretome profiling validated the activation of TLR4 and IL1R signalling by LPS and IL1β ligands and revealed an enrichment in innate immune responses (NF-Kβ, NLRP3, MMPs, Interleukins, etc).Non-toxic doses of BBA [0.5-1000nM] prevented the activation of TLR4 in multiple articular joint cells and inhibited TLR4 & IL1R-dependant innate immune responses at the mRNA and protein level such as inflammatory factors IL6, NOS2, COX2, LCN2, MMP1, -3, -9, -13 and ADAMTS4, among others. Furthermore, NF-Kβ/IKBα and NLRP3/PYCARD/IL1β axis were also severely inhibited after BBA treatment. Moreover, these results were validated by in silico docking analysis that showed BBA interacted with TLR4/NF-Kβ.Conclusion:We prove that BBA inhibit TLR4 & IL1R -dependent innate immune responses in multiple human joint cells (Figure 1). We show that NF-Kβ & NLRP3 signalling, both associated to OA, are blocked (mRNA and protein) after BBA treatment (Figure 1).Our data support previous studies showing the prevention of cartilage loss in an OA animal models by BBA might come from its ability to inhibit TLR4 signalling. In the clinical practice of rheumatologists, Boswellia Serrata could be a useful nutraceutical to manage OA inflammation due to its content in BBA.Figure 1.References:[1]Wang, Q.; Pan, X.; Wong, H.H.; Wagner, C.A.; Lahey, L.J.; Robinson, W.H.; Sokolove, J. Oral and topical boswellic acid attenuates mouse osteoarthritis. Osteoarthr. Cartil.2014, 22, 128–132, doi:10.1016/j.joca.2013.10.012.Acknowledgements:Eloi Franco-Trepat and Ana Lois-Iglesias contributed equally to this work.This research has been funded by the non-profit FER (Fundación Española de Reumatologia /Spanish Foundation of Rheumatology) through the project “Búsqueda de nuevos fármacos bloqueantes de la inflamación asociada a TLR4 en condrocitos humanos artrósicos”.Disclosure of Interests:None declared

Musculoskeletal pathologies (MSPs) such as osteoarthritis (OA) and osteoporosis (OP), are a set of disorders that cause severe pain, motion difficulties, and even permanent disability. In developed countries, the current incidence of MSPs reaches about one in four adults and keeps escalating as a consequence of aging and sedentarism. Interestingly, OA and OP have been closely related to similar risk factors, including aging, metabolic alterations, and inflammation. Visfatin, an adipokine with an inflammatory and catabolic profile, has been associated with several OA and OP metabolic risk factors, such as obesity, insulin resistance, and type II diabetes. Furthermore, visfatin has been associated with the innate immune receptor toll-like receptor 4 (TLR4), which plays a key role in cartilage and bone inflammatory and catabolic responses. Moreover, visfatin has been related to several OA and OP pathologic features. The aim of this work is to bring together basic and clinical data regarding the common role of visfatin in these pathologies and their major shared risk factors. Finally, we discuss the pitfalls of visfatin as a potential biomarker and therapeutic target in both pathologies.

Osteoarthritis (OA), the most common chronic rheumatic disease, is mainly characterized by a progressive degradation of the hyaline articular cartilage, which is essential for correct joint function, lubrication, and resistance. Articular cartilage disturbances lead to joint failure, pain, and disability. Hyaline cartilage is also present in the growth plate and plays a key role in longitudinal bone growth. Alterations of this cartilage by diverse pathologies have been related to longitudinal bone growth inhibition (LBGI), which leads to growth retardation. Diet can play a crucial role in processes involved in the OA and LBGI’s onset and evolution. Specifically, there is ample evidence pointing to the negative impacts of caffeine consumption on hyaline cartilage. However, its effects on these tissues have not been reviewed. Accordingly, in this review, we summarize all current knowledge in the PubMed database about caffeine catabolic effects on articular and growth plate cartilage. Specifically, we focus on the correlation between OA and LBGI with caffeine prenatal or direct exposure. Overall, there is ample evidence indicating that caffeine intake negatively affects the physiology of both articular and growth plate cartilage, increasing consumers predisposition to suffer OA and LBGI. As a result, caffeine consumption should be avoided for these pathologies.

Background The rising prevalence of rheumatic diseases in our society has caused a dramatic impact on the welfare of the population as well as becoming an economic burden to the health system. Osteoarthritis (OA), the most common rheumatic disease, is defined by joint-space narrowing due to progressive cartilage degradation. Despite the growing knowledge in OA pathophysiology, no treatment has yet proved to be efficient enough. The activation of innate immune receptors, such Toll-like receptor 4 (TLR4), by damage-associated molecular patterns (DAMPS) has been involved in chondrocyte-mediated inflammatory responses. There are currently no available drugs aimed to block TLR4-mediated inflammatory responses. Nonetheless, there are already known drugs being employed in other indications that could have this activity; namely amitriptyline, naloxone, and thalidomide. Objectives Determine the ability of amitriptyline, naloxone and thalidomide to block TLR4-mediated innate immune responses in chondrocytes. Methods The effect of amitriptyline, naloxone and thalidomide on TLR4-mediated inflammatory responses was determined in mouse chondrogenic cell line (ATDC5) and in human primary OA chondrocytes. The mRNA expression of key inflammatory factors lipocalin-2 (LCN2), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) was studied by RT-PCR. Cell viability was tested using the methyl-thiazolyl-tetrazolium (MTT) reagent and nitrite accumulation (nitric oxide production) in cell culture media was assessed by Griess reaction and validated by determining nitric oxide synthase 2 (NOS-2) mRNA expression. Results The co-stimulation of human OA chondrocytes with the TLR4 agonist LPS [100 ng/ml] and amitriptyline [1 µM], reduced the mRNA expression of LCN2 (90%), IL-6 (95%) and MCP-1 (87%). The pre-stimulation of naloxone [100 µM] with LPS also reduced the mRNA expression of LCN2 (53%), IL-6 (78%) and MCP-1 (79%). Similar results LCN2 (63%), IL-6 (74%), and MCP-1 (78%) were obtained upon the pre-stimulation of these cells with thalidomide [500 µM]. The anti-inflammatory effect of these drugs on these pro-inflammatory factors was also observed but lowered in ATDC5 cells. Consistent with these results in ATDC5 cells these drugs also reduced the expression of mRNA NOS-2 gene expression as well as nitrite accumulation in the cell culture medium. At the studied concentrations, amitriptyline, naloxone and thalidomide did not affect to chondrocytes viability. Conclusions The data presented here have shown that amitriptyline, naloxone and thalidomide block TLR4 innate immune responses in human OA chondrocytes. These drugs have passed many toxicology and safety tests for their clinical use and could be ready for its repurposing in the management of TLR4-mediated OA cartilage inflammation. Acknowledgements This research is supported by Fondo de Investigacion Sanitaria funded by the Instituto de Salud Carlos III and FEDER (PI16/01870, CP15/00007). R.G. is funded by the Instituto de Salud Carlos III through a Miguel Servet programme. R.G. is a member of the RETICS Programme, RD12/0009/0008 Instituto de Salud Carlos III (ISCIII). Disclosure of Interest None declared