Effect of curcumin on the expression of NOD2 receptor and pro-inflammatory cytokines in fibroblast-like synoviocytes (FLSs) of rheumatoid arthritis (RA) patients.

BACKGROUND: Previous studies has shown that nucleotide-binding and oligomerization domain-containing protein 2 (NOD2) is expressed in Fibroblast-like synoviocytes (FLSs) of rheumatoid arthritis (RA) patients which is stimulated by muramyl dipeptide (MDP) present in the joint environment and induces inflammation via the NF-κB pathway. Also, other studies have shown that curcumin inhibits proliferation, migration, invasion, and Inflammation and on the other hand increases the apoptosis of RA FLSs. In this study, we aim to evaluate the effect of curcumin, a natural anti-inflammatory micronutrient, on the expression of NOD2 and inflammatory cytokines. METHODS: Synovial membranes were collected from ten patients diagnosed with RA and ten individuals with traumatic injuries scheduled for knee surgery. The FLSs were isolated and treated with 40 µM curcumin alone or in combination with 20.3 µM MDP for 24 h. mRNA was extracted, and real-time PCR was performed to quantitatively measure gene expression levels of NOD2, p65, IL-6, TNF-α, and IL-1ß. RESULTS: The study findings indicate that administering MDP alone can significantly increase the mRNA expression levels of IL-6 and IL-1ß in the trauma group and TNF-α in the RA group. Conversely, administering curcumin alone or in combination whit MDP can significantly reduce mRNA expression levels of P65 and IL-6 in FLSs of both groups. Moreover, in FLSs of RA patients, a single curcumin treatment leads to a significant reduction in NOD2 gene expression. CONCLUSION: This study provides preliminary in vitro evidence of the potential benefits of curcumin as a nutritional supplement for RA patients. Despite the limitations of the study being an investigation of the FLSs of RA patients, the results demonstrate that curcumin has an anti-inflammatory effect on NOD2 and NF-κB genes. These findings suggest that curcumin could be a promising approach to relieve symptoms of RA.

Anti-inflammatory effects of PRIMA-1MET (mutant p53 reactivator) induced by inhibition of nuclear factor-κB on rheumatoid arthritis fibroblast-like synoviocytes.

Fibroblast-like synoviocytes (FLSs), the main pathological cells in rheumatoid arthritis (RA), display tumor-like phenotype, including hyper-proliferation, apoptosis resistance, and aggressive phenotype. Excessive proliferation and insufficient apoptosis of RA-FLSs can lead to hyperplastic synovial pannus tissue, excess production of inflammatory mediators, and destruction of joints. In this article, we investigate the effect of PRIMA-1MET on the apoptosis induction and inhibition of pro-inflammatory cytokines in RA-FLSs. Synovial tissue samples were obtained from 10 patients with RA. The FLSs were treated with different concentrations of PRIMA-1MET. The rate of apoptosis and cell survival was assessed by flow cytometry and MTT assay and Real-time quantitative PCR was performed to evaluate the transcription of p53, IL-6, IL-1ß, TNF-α, Noxa, p21, PUMA, Bax, Survivin, and XIAP in treated RA-FLSs. The protein level of p53, IκBα, and phospho-IκBα were measured using Western blotting. The results showed that PRIMA-1MET induced apoptosis in RA-FLSs and increased significantly the expression of Noxa, and decreased significantly IL-6, IL-1ß, p53, and phospho-IκBα expression. PRIMA-1MET can induce apoptosis in RA-FLSs through induction of Noxa expression while p53 was downregulated. Furthermore, PRIMA-1MET treatment results in the suppression of pro-inflammatory cytokine production and NF-κB inhibition. Given the role of p53 and NF-κB in RA-FLSs, PRIMA-1MET can be considered as a new therapeutic strategy for rheumatoid arthritis.

MicroRNA-211-5p Overexpression Effect on Endoplasmic Reticulum Stress and Apoptotic Genes in Fibroblast-like Synoviocytes of Rheumatoid Arthritis.

Fibroblast-like synoviocytes (FLSs) play a major role in the pathogenesis of rheumatoid arthritis (RA). Endoplasmic reticulum (ER) stress and dysregulation of unfolded protein response are involved in the resistance to apoptosis of FLSs in RA (RA-FLSs). MicroRNA (MiR)-211 plays an important role in controlling ER stress and apoptotic genes in a PKR-like ER kinase (PERK)-activating transcription factor 4 (ATF4)-dependent manner. We investigated the effect of miR-211-5p overexpression on ER stress and apoptotic genes in RA-FLSs. FLSs were isolated from synovial tissues of trauma (n=10) and RA (n=10) patients. MiR-211-5p and mRNA expression of the selected genes involved in the PERK pathway and apoptosis regulation were measured in RA, trauma, and thapsigargin (Tg)-treated RA-FLSs. Afterward, Tg-treated RA-FLSs following miR-211-5p overexpression were evaluated for miR-211-5p and mRNA levels of the study genes. The expression of miR-211-5p, PERK, BAX, and BCL2 showed no differences between RA and trauma. However, the expression of ATF4 and BCL-XL showed a significant increase in trauma. In addition, the levels of C/EBP homologous protein (CHOP) and MCL1 indicated a significant increase in RA-FLSs. Tg treatment significantly increased the expression of PERK, ATF4, and CHOP in RA-FLSs with no effect on miR-211-5p, BAX, BCL2, BCL-XL, and MCL1. Furthermore, Tg treatment following miR-211-5p overexpression in RA-FLSs showed a significant increase in levels of miR-211-5p with no changes in apoptotic genes. MiR-211-5p overexpression in stimulated RA-FLSs did not alter the levels of selected genes involved in apoptosis regulation. However, more investigations are necessary to determine the ER stress role in apoptosis regulation in RA-FLSs.

Determination of Normal Reference Values for Meniscal Extrusion Using Ultrasonography During the Different Range of Motion: A Pilot, Feasibility Study.

OBJECTIVES: It is essential to know the normal extrusion measures in order to detect pathological ones. In this study, we aimed to define some normal reference values for meniscal extrusion in the normal knees during different ranges of motion. METHODS: The amount of anterior and posterior portion of meniscal extrusion among 21 asymptomatic volunteers (42 knees) were tracked in 0, 45, and 90° of knee flexion using an ultrasound machine. The repeated measures analysis of variance (ANOVA) was used to show the interaction between the amounts of meniscal extrusion and the different degrees of knee flexion. RESULTS: The anterior portion of the lateral menisci at full knee extension (0.59 ± 1.40) and the posterior portion of the medial menisci during 90° flexion (3.06 ± 2.36) showed the smallest and the highest mean amount of extrusion, respectively. The normal average amounts of anterior extrusion were 1.12 ± 1.17 and 0.99 ± 1.34 mm for medial and lateral menisci, respectively. The posterior meniscal normal extrusions were significantly increasing in both medial and lateral menisci during the survey (F = 20.250 and 11.298; both P values <.001) as they were measured 2.37 ± 2.16 and 1.53 ± 2.18 mm in order. CONCLUSIONS: The medial meniscus can extrude 1.74 ± 1.84 mm normally while this amount was 1.26 ± 1.82 mm for the lateral meniscus. These measures commonly increased with the rising of knee flexion motion. Likewise, the posterior portion showed more extrusion than the anterior portion on both sides. These measures commonly increased with higher knee flexion.

Evaluation of TAK-242 (Resatorvid) Effects on Inflammatory Status of Fibroblast-like Synoviocytes in Rheumatoid Arthritis and Trauma Patients.

Fibroblast-like synoviocytes (FLSs) produce lots of inflammatory molecules that trigger immune responses and intensification the inflammation and thereby play important roles in Rheumatoid Arthritis )RA( pathogenesis. Due to the important roles of toll-like receptor 4 (TLR4) in cytokine production and inflammation, we aimed to evaluate the effects of TAK-242 (Resatorvid) on interleukin (IL)1-ß, IL-6, TNF-α, and TLR4 expression and two important proteins of nuclear factor-κB (NF-κB) signaling pathway (Ikßα and pIkßα) in RA and trauma FLSs. FLSs were isolated from synovial tissues of trauma (n=10) and RA (n=10) patients and cultured in Dulbecco's Modified Eagle Medium (DMEM). 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) was performed to evaluate the cytotoxicity effects of TAK-242 on the RA FLSs. Real-time PCR was performed to measure the expression level of IL1-ß, IL-6, TNF-α, and TLR4 genes in Lipopolysaccharide (LPS) and TAK-242 treated FLSs. Furthermore, the treated FLSs were evaluated for protein levels of Ikßα and pIkßα by western blot. The baseline expression of IL1-ß, IL-6, TNF-α, and TLR4 showed no significant differences between healthy and RA FLSs. LPS stimulated FLSs significantly increased mRNA levels of IL-1ß, IL-6, TNF-α, and TLR4 genes in both the healthy and RA FLSs compared with that of their control groups, and pretreatment with TAK-242 reversed the effect. Furthermore, LPS-stimulated FLSs significantly increased the level of pIkßα in both the healthy and RA FLSs compared with that of their control groups, and pretreatment with TAK-242 reversed the effect. We provide the data that TAK-242 through inhibiting the NF-κB signaling pathway may modulate TLR4-mediated inflammatory responses and could be considered as a potential therapeutic agent for RA patients.

Role of Fibroblast Activation Protein Alpha in Fibroblast-like Synoviocytes of Rheumatoid Arthritis.

Fibroblast-like synoviocytes (FLSs) have been introduced in recent years as a key player in the pathogenesis of rheumatoid arthritis (RA), but the exact mechanisms of their transformation and intracellular pathways have not yet been determined. This study aimed to investigate the role of fibroblast activation protein-alpha (FAP-α) in the regulation of genes involved in the transformation and pathogenic activity of RA FLSs. Synovial FLSs were isolated from RA patients and non-arthritic individuals (n=10 in both groups) and characterized; using immunocytochemistry and flow cytometry analysis. FLSs were divided into un-treated and Talabostat-treated groups to evaluate the FAP-α effect on the selected genes involved in cell cycle regulation (p21, p53, CCND1), apoptosis (Bcl-2, PUMA), and inflammatory and destructive behavior of FLSs (IL-6, TGF-ß1, MMP-2, MMP-9, P2RX7). Gene expression analysis was performed by quantitative real-time polymerase chain reaction (qRT-PCR), and immunoblotting was carried out to evaluate FAP-α protein levels. The basal level of FAP-α protein in RA patients was significantly higher than non-arthritic control individuals. However, no differences were observed between RA and non-arthritic FLSs, at the baseline mRNA levels of all the genes. Talabostat treatment significantly reduced FAP-α protein levels in both RA and non-arthritic FLSs, however, had no effect on mRNA expressions except an upregulated TGF-ß1 expression in non-arthritic FLSs. A significantly higher protein level of FAP-α in FLSs of RA patients compared with that of healthy individuals may point to the pathogenic role of this protein in RA FLSs. However, more investigations are necessary to address the mechanisms mediating the FAP-α pathogenic role in RA FLSs.

An evaluation of the use of a lateral wedged insole and a valgus knee brace in combination in subjects with medial compartment knee osteoarthritis (OA).

Aim: The aim of this study was to evaluate the effect (6 weeks) of the use of a knee brace and a lateral wedge insole, both in isolation and combined, on the knee adduction moment (KAM), pain levels, kinematics (velocity, stride length, cadence), knee ROM, function, and satisfaction in patients with knee osteoarthritis (OA).Methods: Twenty-one patients with medial compartment knee OA were participated in this study. A relatively light three-point valgus knee brace (VB) and full length custom-made lateral wedge insole with arch support were prepared for each subject. Patients divided into three groups at random.Results: The use of a custom fit valgus brace with lateral edge insole concurrently can reduce a greater peak KAM than use in isolation (30%). In addition, the use of both interventions can improve walking velocity, cadence, and reduce levels of pain in patients with medial compartment OA.Conclusion: All parameters except stride length and KAM in comparison with first day of wearing interventions improved significantly in the combined, concurrent usage group. The use of a lateral wedge insole and a valgus knee brace in combination can improve the kinetic and kinematic parameters in patients with medial compartment knee OA.

Safety and efficacy of engineered tissue composed of silk fibroin/collagen and autologous chondrocytes in two patients with cartilage defects: A pilot clinical trial study.

BACKGROUND: The objective of this pilot clinical trial study was to evaluate safety and effectiveness of the newly engineered tissue composed of autologous chondrocytes and collagen/fibroin scaffold in repair of osteochondral defects. METHODS: We implemented a pilot clinical study in two patients with knee osteochondral lesions using engineered tissue composed of scaffold and autologous chondrocytes. Patients were clinically evaluated using the International Repair Cartilage Society score and magnetic resonance imaging (MRI) for one year. RESULTS: Improved clinical outcomes and objective scores indicated a normal or nearly normal knee in both patients. International Knee Documentation Committee score was upgraded from 34.5 at baseline to 72.4 in the first patient, and 28.7 to 81.6 in the second patient. Visual analogue scale, showing the suffering pain score, was lowered from 8 to 0 in both patients, Western Ontario and McMaster Universities Osteoarthritis Index score representing the physical ability of the patients was changed from 68.1 to 87.1 in Patient 1 and 58.3 to 87.1 in Patient 2, the knee function score, related to the functional ability of the knee, was improved from 70 to 100 in the first patient and from 45 to 91 in the second patient. MRI showed great coverage and integration of the graft in patients, with no effusion, decreased edema and cartilage formation signals. CONCLUSIONS: The functional and clinical outcomes alongside MRI data showed promising results for regenerating osteochondral defects. A randomized clinical trial study is required to confirm feasibility of this novel engineered tissue in repair of osteochondral defects.

Two-stage Total Knee Arthroplasty for Treatment of Surgical Failure of Septic Arthritis in Degenerative Knee Joints.

BACKGROUND: The ultimate goal of the treatment of infectious knee arthritis is to protect the articular cartilage from adverse effects of infection. Treatment, however, is not always hundred percent successful and has a 12% failure rate. Persistent infection is more likely to happen in elderly patients and those with underlying joint diseases, particularly osteoarthritis. Eradication of infection and restoration of function in the involved joint usually are not possible by conventional treatment strategies. There are few case series reporting two-stage primary knee arthroplasty as the salvage treatment of the septic degenerative knee joint; however, the treatment protocol remains to be elucidated. METHODS: Based on a proposed approach, patients with failure of common interventions for treatment of septic knee arthritis and underlying joint degeneration were treated by two-stage TKA and intervening antibiotic loaded static cement spacer. Suppressive antibiotic therapy was not prescribed after the second stage. RESULTS: Complete infection eradication was achieved with mean follow up of 26 months. All cases were balanced with primary total knee prosthesis. The knee scores and final range of motions were comparable to other studies. CONCLUSION: The two-stage total knee replacement technique is a good option for management of failure of previous surgical treatment in patients with septic arthritis and concomitant joint degeneration. Our proposed approach enabled us to use primary prosthesis in all of our patients with no need for suppressive antibiotic therapy.

Loop-Post Construct, A Novel Technique for Medial Meniscus Root Repair.

Surgical reattachment of medial meniscus posterior root tear (MMPRT) with transtibial sutures can delay the presence of medial knee joint compartment osteoarthritis. Most suture configurations are placed five mm away from the torn margin in the meniscal substance which is already degenerated and may decrease the pull out strengths of repair construct. The number of meniscus penetration may also be important considering meniscus tissue damage with more complex suture techniques impose the risk of suture cut out through the meniscus substance. We introduce our loop postsuture construct technique which is simple, cheap and reproducible.

Epigenetics in rheumatoid arthritis; fibroblast-like synoviocytes as an emerging paradigm in the pathogenesis of the disease.

Rheumatoid arthritis (RA) is characterized by immune dysfunctions and chronic inflammation that mainly affects diarthrodial joints. Genetics has long been surveyed in searching for the etiopathogenesis of the disease and partially clarified the conundrums within this context. Epigenetic alterations, such as DNA methylation, histone modifications, and noncoding RNAs, which have been considered to be involved in RA pathogenesis, likely explain the nongenetic risk factors. Epigenetic modifications may influence RA through fibroblast-like synoviocytes (FLSs). It has been shown that FLSs play an essential role in the onset and exacerbation of RA, and therefore, they may illustrate some aspects of RA pathogenesis. These cells exhibit a unique DNA methylation profile in the early stage of the disease that changes with disease progression. Histone acetylation profile in RA FLSs is disrupted through the imbalance of histone acetyltransferases and histone deacetylase activity. Furthermore, dysregulation of microRNAs (miRNAs) is immense. Most of these miRNAs have shown an aberrant expression in FLSs that are involved in proliferation and cytokine production. Besides, dysregulation of long noncoding RNAs in FLSs has been revealed and attributed to RA pathogenesis. Further investigations are needed to get a better view of epigenetic alterations and their interactions. We also discuss the role of these epigenetic alterations in RA pathogenesis and their therapeutic potential.

Arthroscopic ablation of an osteoid osteoma of the elbow: a case series with a minimum of 18 months' follow-up.

BACKGROUND: Arthroscopic excision of an osteoid osteoma was first reported in the knee joint; since then, there have been several reports of arthroscopic excisions in the knee, shoulder, and elbow, with inconclusive outcomes because of a limited number of cases. The aim of this prospective study was to evaluate the medium-term functional effects of arthroscopic ablation in cases of an osteoid osteoma around the elbow. METHODS: We treated osteoid osteoma of the elbow through arthroscopic ablation in 10 patients. The arthroscopic resection procedure was performed 23 ± 9 months (range, 12-36 months) after initial symptoms. At the preoperative examination and last follow-up examination, the elbow flexion-extension and forearm supination-pronation ranges of motion were measured. The patients were assessed by the Mayo Elbow Performance Score, the visual analog scale for the elbow and wrist, and the Quick Disabilities of the Arm, Shoulder, and Hand score. Finally, the patients' general satisfaction was assessed. RESULTS: The postoperative elbow flexion-extension range of motion was significantly higher compared with range of motion before surgery (P = .001; r = 0.86). According to the Mayo Elbow Performance Score, the average score increased significantly at the final follow-up examination. The mean preoperative and final Quick Disabilities of the Arm, Shoulder, and Hand scores were 47 ± 14 and 1.6 ± 2.8, respectively (P < .001). All patients were satisfied with the operation result. CONCLUSION: According to the results of our study, arthroscopic ablation is a safe and efficient method of treatment for osteoid osteoma of the elbow, with a fast rehabilitation time.

Surgical treatment of pectoralis major tendon rupture (outcome assessment).

PURPOSE: Rupture of pectoralis major (PM) occurs most commonly as a result of an indirect mechanism associated with extensive tension on a maximally contracted muscle. Patients with PM tendon ruptures, classically present a history of sudden severe pain in arm and shoulder at the time of injury. Treatment options vary from conservative to operative. In cases with total or near-total injuries, surgical treatment by anatomic repair is generally advised, since conservative treatment may lead to poor results. The present paper reports 24 cases of surgically-treated ruptured PM while assessing the results. METHODS: Between 2005 and 2010, 32 cases of unilaterally distal ruptured PM were surgically treated by the same surgeon and same technique, in two teaching hospitals of Tehran University of Medical Sciences. All cases were followed postoperatively by physical examination and functional criteria. RESULTS: Since eight of the cases were lost from follow up, 24 cases were followed for at least one year. According to modified Kakwani system, 6 of our patients (25%) rated as excellent, 15 cases(62.5%) rated as good, 2 cases (8.33%) rated as fair and 1 case (4.1%) rated as poor. CONCLUSION: In conclusion, we see that 87.5% of the patients had good to excellent results, according to modified Kakwani classification, after one year of follow up. So it seems that surgical repair of the pectoralis major ruptures will help the patients to return to their previous activities more frequently and we believe, to achieve better functional outcome. Meticulous surgical technique and attention to rehabilitation program are more important than delay in surgery.