A Study on the Treatment of Rheumatoid Arthritis Using a Novel GelMA-HAMA Dual-Network Hydrogel Microneedle Loaded with MTX-NCs in Combination with Adalimumab.

In this work, the expression of secreted frizzled related protein 2 (SFRP2) in rheumatoid arthritis (RA) model rats and the mechanisms of SFRP2 on the RA pathogenesis were investigated. Data suggested that SFRP2 was significantly down-regulated in RA model rats compared with normal control, and overexpression of SFRP2 suppressed the RA pathogenesis and the canonical Wnt signaling in fibroblast-like synovial cells (FLS) from RA model rats, whereas knockdown of SFRP2 got an opposite observation. Interestingly, 5-azadC treatment up-regulated the SFRP2 expression, inhibited the FLS proliferation, suppressed the expression of IL-6 and IL-8 and the fibronectin production, suggesting that the decreased SFRP2 in RA model rats was due to the DNA methylation. Furthermore, DNMT1 knockdown up-regulated the SFRP2 expression, DNMT1 overexpression inhibited the SFRP2, and the quantitative methylation-specific PCR (qMSP) confirmed that the DNMT1 has direct methylation roles for the SFRP2 promoter, leading to a regulation of FLS proliferation and fibronectin expression in RA model rats. In addition, up-regulated MeCP2 was involved in the SFRP2 regulation and the pathogenesis of RA model rats, and MeCP2 and DNMT1 have synergistic inhibition roles in the SFRP2 expression. Combination of DNMT1 and DNA methylation may be a promising treatment strategy for individuals with RA in which SFRP2 is down-regulated.

Fibroblast-like synoviocytes (FLSs) are a key cell type involved in rheumatoid arthritis (RA) progression. We previously identified the KCa1.1 potassium channel (Maxi-K, BK, Slo 1, KCNMA1) as a regulator of FLSs and found that KCa1.1 inhibition reduces disease severity in RA animal models. However, systemic KCa1.1 block causes multiple side effects. In this study, we aimed to determine whether the KCa1.1 β1-3-specific venom peptide blocker iberiotoxin (IbTX) reduces disease severity in animal models of RA without inducing major side effects. We used immunohistochemistry to identify IbTX-sensitive KCa1.1 subunits in joints of rats with a model of RA. Patch-clamp and functional assays were used to determine whether IbTX can regulate FLSs through targeting KCa1.1. We then tested the efficacy of IbTX in ameliorating disease in two rat models of RA. Finally, we determined whether IbTX causes side effects including incontinence or tremors in rats, compared with those treated with the small-molecule KCa1.1 blocker paxilline. IbTX-sensitive subunits of KCa1.1 were expressed by FLSs in joints of rats with experimental arthritis. IbTX inhibited KCa1.1 channels expressed by FLSs from patients with RA and by FLSs from rat models of RA and reduced FLS invasiveness. IbTX significantly reduced disease severity in two rat models of RA. Unlike paxilline, IbTX did not induce tremors or incontinence in rats. Overall, IbTX inhibited KCa1.1 channels on FLSs and treated rat models of RA without inducing side effects associated with nonspecific KCa1.1 blockade and could become the basis for the development of a new treatment of RA.

Interleukin (IL)-1β serves a crucial role in the progression of rheumatoid arthritis. Previous studies have indicated that the ERK/STAT1 signaling pathway may be involved in the inflammatory response in synovial fluid-derived fibroblast-like synoviocytes (sfd-FLSs). However, the molecular mechanisms underlying the pathological effects of the inflammatory factors induced by IL-1β in sfd-FLSs remain unclear. The aim of the present study was to investigate the IL-1β-mediated signaling pathways involved in the expression of inflammatory factors in sfd-FLSs and in a rat model of rheumatoid arthritis. Reverse transcription-quantitative PCR, western blotting, and immunohistochemistry were used to analyze the role of IL-1β in the rat model of rheumatoid arthritis. The results suggested that IL-1β administration exacerbated rheumatoid arthritis, bone injury and increased the expression levels of inflammatory factors, such as IL-17 and tumor necrosis factor α (TNF-α), whereas treatment with anti-IL-1β exhibited opposite effects. In vitro experiments in sfd-FLSs further suggested that treatment with IL-1β influenced the expression levels of various inflammatory factors. In specific, IL-1β increased the expression of IL-17 and TNF-α, and decreased the expression of IL-6 and IL-10 in sfd-FLSs. Additionally, treatment with IL-1β increased the mRNA expression and protein phosphorylation of NF-κB, ERK and STAT1 in sfd-FLSs. Treatment with anti-IL-1β exhibited opposite effects on the expression levels of inflammatory factors and suppressed the NF-κB-mediated ERK-STAT1 signaling pathway activation in sfd-FLSs. Finally, treatment with a NF-κB inhibitor suppressed the effects of IL-1β, and NF-κB overexpression reversed the effects of anti-IL-1β on the expression levels of IL-17, TNF-α, NF-κB, ERK and STAT1. In conclusion, the present results demonstrated that treatment with IL-1β increased the expression levels of inflammatory factors in sfd-FLSs via the regulation of the NF-κB-mediated ERK/STAT1 signaling pathway in a rat model of rheumatoid arthritis. Therefore, the NF-κB/ERK/STAT1 signaling pathway may represent a potential target for the development of novel treatments for rheumatoid arthritis.

Qufeng epimedium decoction alleviates rheumatoid arthritis through CYLD-antagonized NF-kB activation by deubiquitinating Sirt1.

The present study aimed to construct a rat model of rheumatoid arthritis (RA) to evaluate changes in pathology, the expression of inflammatory factors and regulation of signaling pathways. The protective effect of tetrandrine (Tet) on tissue lesions induced by RA was also investigated. A total of 60 Wistar rats (100-200 g) were randomly divided into six groups (n=10 per group), namely a blank (NC) group, model group, methotrexate (MTX) group (3 mg/kg body weight), high-dose Tet group (31.25 mg/kg body weight), medium-dose Tet group (18.75 mg/kg body weight) and low-dose Tet group (6.25 mg/kg body weight). A rat model of RA was induced via injection of 0.1 ml complete Freund's adjuvant into the right rear toe. Toe swelling rate, arthritis index and immune organ index were calculated. In addition, cyclooxygenase (COX)-2 expression at the mRNA and protein level in the peripheral blood mononuclear cells (PBMCs) of rats were determined by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Serum concentrations of inflammatory factors were measured using enzyme-linked immunosorbent assays. It was observed that treatment with Tet alleviated the severity of rear toe swelling associated with RA in rats. Furthermore, Tet exerted anti-inflammatory and immunosuppressive effects in the rat model of RA. Tet also reduced the expression of COX-2 in PBMCs and lowered the concentrations of inflammatory factors in the serum of RA rats. The present data indicate that Tet may exert pharmacological effects in the treatment of RA. The mechanism of action of Tet may be associated with the regulation of inflammatory factors and the inhibition of immune organs.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease for which treatment focuses on suppressing an overactive immune system and maintaining the physiological balance of synovial fibroblasts (SFs). We found that miR-30-5p was highly expressed in rheumatoid arthritis synovial fibroblasts (RASFs). Subsequently, we predicted that phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) might be a putative target of miR-30-5p. Recent studies have reported that PIK3R2 can maintain the physiological homeostasis of RASFs. Therefore, miR-30-5p inhibitor has the potential to be used in the treatment of RA, but low levels of miR-30-5p inhibitor internalization affect its application. Triptolide (TP) is an effective drug in the treatment of RA but induces severe toxicity and has a narrow therapeutic window. In this study, the cell internalization performance of miR-30-5p inhibitor was improved by loading it into cell membrane penetrating peptide (CADY)-modified mesoporous silica nanoparticles (MSNs), and the toxicity of TP was decreased by loading it into a controlled drug release system based on MSNs. The nanodrug carrier was constructed by filling a phase-change material (PCM) of 1-tetradecanol and drugs into MSNs that could be triggered by an NIR laser with thermo-chemo combination RA therapy. Our results show that the miR-30-5p inhibitor-loaded MSNs@CADY significantly inhibited RASF proliferation and increased apoptosis. In addition, MSNs@PCM@TP under 808 nm laser irradiation were effective in downregulating immune system activation in an RA rat model. Finally, the results of a pharmacodynamics study showed that the combination of MSNs@CADY@miR-30-5p inhibitor and MSNs@PCM@TP under 808 nm laser significantly increased the effectiveness of RA treatment. These findings provide a novel understanding of RA pathogenesis and a theoretical basis for RA treatment.

Ipomoea pes-caprae (L.) Sweet (Convolvulaceae) is a commonly used marine Chinese medicine in the coastal areas of southern China. Traditionally, it has been used in the treatment of rheumatoid arthritis (RA). However, the mechanism of action against RA remains unclear. This study aimed to explore the mechanism of action of Ipomoea pes-caprae water extract (IPE) in the treatment of RA through serum metabolomics and network pharmacology. Rat models of RA with wind-dampness cold bi-syndrome (WCM) and wind-dampness heat bi-syndrome (WHM) were established to evaluate the therapeutic effect of IPE against RA. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) technology was used to analyze the absorbed components of IPE in the plasma of the two models. Serum metabolomics was employed to identify potential biomarkers and metabolic pathways of IPE in the treatment of RA. The key targets and related pathways of RA were screened using network pharmacology and validated using molecular docking. The biomarker-pathway-target network was mapped via the combination of metabolomics and network pharmacology. A total of 10 chemical constituents were identified from WHM rat plasma, and eight chemical constituents were identified from WCM rat plasma. Serum metabolomics research identified 20 endogenous potential biomarkers, and 10 major metabolic pathways closely related to WHM and WCM. Network pharmacology analysis yielded 65 overlapping targets, with the core targets being ALB, AKT1, EGFR, and CASP3. Molecular docking showed that the four absorbed components in plasma had a strong binding activity with ALB and AKT1. Combining metabolomics and network pharmacology, two major biomarkers and two major pathways were identified. IPE can effectively relieve the symptoms of RA, and the potential mechanism of IPE in treating RA has been preliminarily elucidated. These results can provide a scientific basis for further drug research and development, as well as clinical application.

Introduction: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease. It is characterized by a persistent synovitis, progressively causing cartilage destruction and bone erosion, leading to joint deformation and disability. RA patients may have highly variable patterns of radiologic progression over time. In any case, joint damage and functional status loss appear since the early disease stages. RA can potentially involve other organs, causing, for instance, severe respiratory and/or cardiovascular complications; for this reason, a higher mortality risk (as compared with general population) is associated with RA. Biological disease-modifying antirheumatic drugs are recommended for use particularly in rheumatoid arthritis patients who had an inadequate response to methotrexate. Objectives: Using data on efficacy from the RA-BEAM trial (American College of Rheumatology (ACR) indices) and number needed to treat (NNT) (Rovasio et al 2018), the present economic analysis compared the cost per NNT (ACR 20, 50 and 70) on week 24 of Baricitinib (BARI) (4mg) and Adalimumab (ADA) (40mg) in adult patients with moderately to severely active RA with an inadequate response to methotrexate (MTX) in Brazil (R$, local currency). In addition to that, the direct treatment cost on weeks 12, 24 and 52 was also compared. Methods: The economic analysis for both biologic DMARDs (BARI 4mg and ADA 40mg) was developed considering the number of doses on each label with 12, 24 and 52 weeks of treatment; NNT on week 24 according to Rovasio et al 2018; drug acquisition cost using as reference the Brasíndice pharmaceutical guide (wholesale price + 18% taxes); the cost of MTX was not considered. Results: BARI presented lower treatment cost compared to ADA on weeks 12, 24 and 52, demonstrating 48% of reduction. BARI also presented lower NNT compared to ADA on week 24. The cost per responder considering ACR (20, 50 and 70) for BARI was lower compared to ADA (ACR20: R$66.915 vs R$162.096; ACR50: R$79.850 vs R$183.741; ACR70: R$114.178 vs R$348.723, respectively). Conclusion: The results from this economic evaluation, based on the cost per NNT (Rovasio et al 2018) in combination with drug acquisition cost (Brasíndice pharmaceutical guide), showed that BARI is a more cost-effective therapeutic option when compared with ADA in the treatment of moderately to severely active RA in adult patients with an inadequate response to MTX.

Development, characterization, and optimization of folate-chitosan surface-modified PLGA-decorated mesoporous silica nanoparticles for effective delivery of leflunomide for the management of rheumatoid arthritis.

Background: Biological therapy is indicated in the treatment of RA (Rheumatoid Arthritis) after failure of disease-modifying anti-rheumatic drugs (DMARDS) by the ACR/EULAR recommendations. The objective of the study is to describe the characteristics of Saudi patients at the initiation of biological therapy and to evaluate clinical effectiveness of this therapy measured by the disease activity score DAS 28. Methods: This was a retrospective cohort study of RA (rheumatoid arthritis) patients in King Fahad Hospital in Jeddah, Saudi Arabia from January 2005-July 2011. Data were collected from the medical records of all RA patients on biological therapy including: demographics, disease characteristics, comorbid illnesses and DAS 28 score over a period of 1 year. Results: 139 patients were studied (mean age 46 ± 13 years), of which 118 (84%) were females; mean duration of affliction with RA was 7.2 years ranging 1-45 years. Rheumatoid factor (RF) was positive in 88 patients (63.3%) and one or more comorbidities were present in 102 patients (73.3%). First choice of biological drug was ADA (Adalimumab) 44 patients (31.7%) and RTX (Rituximab) was the 2nd frequently prescribed biological drug. Mean DAS 28 activity at baseline was in ADA 41 patients (6.10 ± 1.62), ETA (Etarnercept) 29 patients (6.64 ± 1.42) and RTX 50 patients (6.7 ± 1.32). Moderate to good EULAR response was obtained in 74%, 85.7% and 53.3% at 6 months in ADA, ETA and RTX patients respectively. Moderate to good EULAR response was obtained in 61.8%, 86.6% and 72% in ADA, ETA AND RTX patients respectively at 1 year of treatment. Therapeutic effectiveness was comparable with the response rates in published observational trials. Conclusion: Our data demonstrate daily clinical practice in management of RA. The pattern of prescription is in agreement with the ACR/EULAR recommendations for initiation of biologicals in the treatment of RA.

To the Editor, Bone erosion is the most important radiographic feature of rheumatoid arthritis (RA) and has been considered as almost irreversible. Although biologics has improved the clinical outcomes of RA, radiographic repair of bone erosions is still limited [1]. Tocilizumab (TCZ), an anti-interleukin-6-receptor antibody, has been shown to be highly efficacious in the treatment of RA, regardless of the age of patients, and to inhibit progression of joint damage [2–4]. Here we present the case of a very elderly patient with RA who showed dramatic radiographic repair by TCZ. An 80-year-old woman was referred to Hikarigaoka Spellman Hospital for treatment of active RA. She had been diagnosed with RA 4 years earlier (rheumatoid factor, 182 IU/ml) and had been treated with methotrexate (6 mg/week) and salazosulfapyridine. The patient had a swollen joint count (SJC) of 6 and tender joint count (TJC) of 6. The patient global assessment using a visual analogue scale was 79 mm. Erythrocyte sedimentation rate (ESR) was 29 mm/h and anti-cyclic citrullinated peptide (CCP) antibody was above 100 U/ml. The 28-joint disease activity score using ESR (DAS28-ESR) was 5.52. Steinbrocker functional class was II and stage was IV. After administration of TCZ (8 mg/kg every 4 weeks) as monotherapy, she achieved DAS28-ESR remission by 16 weeks and Boolean remission by 34 weeks. Since then, she has been in remission for 3 years with no serious adverse events except for one incidence of acute pneumonia caused by Chlamydia pneumoniae. Follow-up X-rays for 3 years showed dramatic repair of bone erosion (Figure 1a–h). The scale of erosions had decreased and a smooth cortex had been generated on the surface of the erosions. The van der Heijde-modified total Sharp score (mTSS), which was scored by two trained readers in a blinded manner, also improved year by year (Figure 1i). Figure 1. Metacarpophalangeal (MCP) joint of left index finger at first visit (a) and after 1 year (b), 2 years (c) and 3 years (d). The swelling of the joint disappeared by 3 weeks after administration of TCZ. MCP joint of right index finger at first visit (e) ... Radiographic repair does occur, but it is observed almost exclusively in joints with no signs of swelling [5]. van der Linden et al. reported that radiographic repair was observed in 18 out of 250 patients (7.2%), and patients who showed radiographic repair had a high prevalence of autoantibodies such as rheumatoid factor and anti-CCP antibody [6]. It has also been shown in an animal model that when synovitis resolves, osteoblasts migrate to the surfaces of eroded bone, resulting in new bone formation [7]. These studies indicate that complete resolution of inflammation at the sites of erosions is essential in repair of bone erosions. The characteristics of elderly onset RA (EORA) that is diagnosed at above 60 years of age differ from young onset RA (YORA) by a more equal sex distribution, a higher disease activity and a higher frequency of large joint involvements [8]. In the cases of seropositive patients, more radiographic damage and functional decline have been observed in EORA patients than in YORA patients [9]. In EORA patients, age-associated factors, such as comorbidities and decreased drug metabolism capacity, often limit the treatment options and worsen their prognosis [10, 11]. We previously reported that no significant difference in efficacy or safety profile of TCZ was found between the groups of elderly (over 65 years old) and nonelderly (below 65 years old) patients and that SJC < 1 was achieved by 67% of patients after 6 months of TCZ use [4, 12]. TCZ enables both rapid reduction of joint swelling and long-term complete remission irrespective of age, leading to repair of bone erosions. In fact, this case indicates that bone repair can occur even in a very elderly patient with RA during long-term complete remission with TCZ.

AB0191 DECREASING DELAY TO DIAGNOSIS AND TREATMENT OF RHEUMATOID ARTHRITIS: STILL DIFFICULT TO TREAT WITHIN THE WINDOW OF OPPORTUNITY

Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by persistent joint inflammation and pain. Conventional systemic therapies, typically administered orally or by injection, often result in gastrointestinal side effects and poor patient compliance. Although microneedles (MN) enable localized and minimally invasive drug delivery through the skin, their therapeutic efficacy remains limited by uncontrollable and unpredictable release kinetics. Here, this work reports a wearable electro-controlled MN device (EMND) for synergistic RA therapy. The EMND mainly consists of a MN array, a conductive hydrogel, a miniature control circuit and a smartphone-based application. The MN array facilitates transdermal access at the lesion site, while the conductive hydrogel enables electrically triggered, on-demand drug release via iontophoresis. This design allows precise spatiotemporal control of multiple therapeutics to achieve synergistic anti-inflammatory and analgesic effects. In RA rat models, EMND treatment reduced inflammatory cytokine levels to 11.38%-14.51% of baseline and increased pain thresholds by 5.24-fold. Overall, this wearable EMND offers an exciting prospect for facilitating RA treatments and autonomy of therapeutic interventions, with potential applications for other chronic conditions.

Hyaluronic acid microneedle patch for transdermal delivery of naringin-loaded PEGylated terpesomes in a rat model of rheumatoid arthritis: Modulation TGF-β1, oxidative stress, and inflammation.

Rheumatoid arthritis (RA) can lead to joint destruction and deformity, which is a significant cause of the loss of the young and middle-aged labor force. However, the treatment of RA is still filled with challenges. Though dexamethasone, one of the glucocorticoids, is commonly used in the treatment of RA, its clinical use is limited because of the required high-dose and long-term use, unsatisfactory therapeutic effects, and various side-effects. Ultrasound-targeted microbubble destruction (UTMD) can augment the ultrasonic cavitation effects and trigger drug release from targeted nanocarriers in the synovial cavity, which makes it a more effective synergistic treatment strategy for RA. In this work, we aim to utilize the UTMD effect to augment the synergistic therapy of RA by using polyethylene glycol (PEG)-modified folate (FA)-conjugated liposomes (LPs) loaded with dexamethasone sodium phosphate (DexSP) (DexSP@LPs-PEG-FA). The UTMD-mediated DexSP@LPs-PEG-FA for targeted delivery of DexSP including a synergistic ultrasonic cavitation effect and drug therapy were investigated through in vitro RAW264.7 cell experiments and in vivo collagen-induced arthritis SD rat model animal experiments. The results show the DexSP release from targeted liposomes was improved under the UTMD effect. Likewise, the folate-conjugated liposomes displayed targeting association to RAW264.7 cells. Together with the application of ultrasound and microbubbles, liposomes-delivered DexSP potently reduced joints swelling, bone erosion, and inflammation in both joints and serum with a low dose. These results demonstrated that UTMD-mediated folate-conjugated liposomes are not only a promising method for targeted synergistic treatment of RA but also may show high potential for serving as nanomedicines for many other biomedical fields.