interleukin-8 and Osteoarthritis--Knee

Allergy against implant materials is discussed controversially and still not fully understood. Despite these controversies, a relevant number of patients receive hypoallergenic knee implants. The aim of this study was to compare a new coating system with the standard implant in total knee arthroplasty (TKA). Additionally, the influence of proinflammatory cytokines on patient-reported outcome measures (PROMs) was investigated.. 120 patients without known metal allergy and without previous metal implants were included. The patients were randomized to receive a coated or standard TKA of the same knee system. 105 patients completed the 5 year follow-up. Patient-reported outcome measures (PROMs) including knee function (Oxford Knee Score, OKS), quality of life (SF36) and UCLA activity scale were assessed. Additionally, several cytokines with a possible role in implant allergy were measured in patient`s serum (IL-1beta, IL-5, IL-6, IL-8, IL-10, IP-10, IFN γ, TNF α). Group comparison was performed using Mann-Whitney U test for continuous values and chi-square test for categorical values.. There were no differences in PROMs between both groups at any follow-up. The majority of patients demonstrated no elevation of the measured blood cytokines. The blood cytokine pattern after 5 years demonstrated no differences between study groups. There was a significant association between elevated IL-8 values and worse results in the overall OKS (p = 0.041), the OKS function component (p = 0.004), the UCLA activity scale (p = 0.007) and the physical component of SF36 (p = 0.001).. There were no problems with the new coating during mid-term follow-up and no differences in PROMs between coated and standard TKA. Patients with an increased inflammatory response demonstrated worse functional results, regardless of the implant.. I.. The study protocol was registered in the US National Institutes of Health's database ( http://www.. gov ) registry under NCT00862511.

Topics: Arthroplasty, Replacement, Knee; Cytokines; Humans; Hypersensitivity; Interleukin-8; Knee Joint; Knee Prosthesis; Osteoarthritis, Knee; Patient Reported Outcome Measures; Quality of Life; Treatment Outcome

Although the exact etiology of patient dissatisfaction in total knee arthroplasty (TKA) is unclear, the inflammatory response precipitated by surgery may be implicated. Robotic TKA has been shown to result in reduced bone and soft-tissue trauma. The objectives of this study were to compare the inflammatory response in conventional jig-based TKA versus robotic-arm-assisted TKA and to examine the relationship with early functional outcomes.. This prospective randomized controlled trial included 15 patients with symptomatic knee osteoarthritis undergoing conventional TKA and 15 undergoing robotic-arm-assisted TKA. Blood samples were collected for up to 28 days postoperatively, and predefined markers of systemic inflammation were measured in serum. The local inflammatory response was assessed by analyzing samples from the intra-articular drain fluid at 6 and 24 hours. Relationships with early functional outcomes were evaluated using the Spearman rank correlation coefficient.. Patients in the robotic TKA group demonstrated lower levels of interleukin (IL)-6 in the drain fluid at 6 hours (798.54 pg/mL versus 5,699.2 pg/mL, p = 0.026) and 24 hours and IL-8 at 6 hours. Robotic TKA was associated with lower pain scores on postoperative days 1, 2, and 7. Patient-reported outcome measures were comparable between the 2 groups at 2 years. Significant correlations were observed between all serum markers except IL-1b and self-reported pain on postoperative day 7; between drain IL-8 levels and pain on postoperative days 1 (r = 0.458), 2, and 7; and between drain IL-6, IL-8, and tumor necrosis factor-alpha levels at 6 hours and knee flexion or extension.. Robotic-arm-assisted TKA was associated with a reduction in the early postoperative local inflammatory response. We also found a moderate relationship between the inflammatory responses and self-reported pain, knee flexion, and knee extension. Further validation of these findings on a larger scale and using longer-term outcomes will be key to developing the optimal TKA procedure.. Therapeutic Level I . See Instructions for Authors for a complete description of levels of evidence.

Topics: Arm; Arthroplasty, Replacement, Knee; Humans; Interleukin-8; Knee Joint; Osteoarthritis, Knee; Pain; Prospective Studies; Robotic Surgical Procedures

The aim of this study was to evaluate the effect of an oral preparation containing a naturally occurring matrix of hydrolyzed collagen type II, chondroitin sulfate (CS), and hyaluronic acid (HA), and bioactive oligopeptides of natural hydrolyzed keratin (K) in patients affected by knee OA through the evaluation of synovial fluid (SF) and clinical changes before and after treatment. Thirty patients with knee OA and swollen joint were included in the study and submitted to arthrocentesis. Patients were randomized in two groups: 1) the treatment group (N.15) took a dietary supplement containing 120 mg HA, 240 mg CS and 300 mg K once a day for 4 weeks; 2) the control group (N.15) was only submitted to arthrocentesis. Patient symptoms were evaluated at the beginning and at the end of the study by the WOMAC self-assessment questionnaire, the Lequesne algofunctional index, and the VAS forms. SF changes were evaluated by measuring local inflammatory indices, cytokines IL-1β, IL-8, IL-6, IL-10 and GM-CSF. The group of patients treated with the oral supplement showed an improvement in the clinical indices WOMAC (p<0.01), Lequesne (p=0.014) and VAS pain (p<0.01). On the contrary, no significant changes were found in the control group. The SF collected from the treated group showed a reduction of IL-8 (p=0.015), IL-6 and IL-10 levels, while no changes in cytokines were observed in the control group. This pilot study suggests that an oral administration of a preparation containing a combination of HA, CS and K can improve some clinical parameters and affect cytokine concentrations in SF in patients with knee OA.

Topics: Administration, Oral; Arthrocentesis; Chondroitin Sulfates; Collagen Type II; Drug Combinations; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hyaluronic Acid; Interleukin-10; Interleukin-1beta; Interleukin-6; Interleukin-8; Keratins; Middle Aged; Osteoarthritis, Knee; Pilot Projects; Symptom Assessment; Synovial Fluid

This study was designed to investigate the analgesic effects of nimesulide and celecoxib in patients with knee osteoarthritis (OA). In patients with joint effusion, the effects of these non-steroidal anti-inflammatory drugs (NSAIDs) on synovial fluid concentrations of substance P (SP), interleukin (IL)-6 and IL-8 also were evaluated.. Patients were randomly assigned either nimesulide (100 mg twice a day) or celecoxib (200 mg once a day) for 2 weeks. The intensity of joint pain was assessed with a 100-mm visual analogue scale (VAS). Furthermore, patients completed questions about analgesic efficacy and overall tolerability of the treatments on a five-point categorical scale. Synovial fluid samples were drawn at baseline, 30 min after the first drug intake (day 1), and 30 min after the last drug intake (day 14).. We enrolled 44 patients, 20 of whom had a joint effusion. In this group, the effects of nimesulide were more marked than for celecoxib, with evidence of a faster onset of the analgesic action. Both after a single or repeated administration, nimesulide significantly reduced the synovial fluid concentrations of SP and IL-6. Celecoxib, on the other hand, did not change the concentrations of SP and significantly reduced the levels of IL-6 only on day 14. None of the drugs affected IL-8. Both drugs were generally well tolerated.. These results provide evidence that nimesulide is an effective agent for the symptomatic treatment of OA. The effect on inflammatory pain mediators is consistent with the fast analgesic action of this NSAID.

Topics: Aged; Anti-Inflammatory Agents, Non-Steroidal; Arthralgia; Celecoxib; Double-Blind Method; Female; Humans; Interleukin-6; Interleukin-8; Male; Osteoarthritis, Knee; Pain Measurement; Pyrazoles; Substance P; Sulfonamides; Synovial Fluid; Treatment Outcome

Our aim was to investigate the effects of hyaluronan on inflammatory cytokines in the synovial fluid of patients with knee osteoarthritis. The study was single blind, placebo-controlled, and randomized. We administered hyaluronan to 22 patients in the study group and placebo to 19 in the control group. Enzyme-linked immunosorbent assay was used to determine the levels of cytokines. Both HA and placebo caused a significant decrease in interleukin (IL)-6 levels (P=0.0001 and P=0.04, respectively). But it was more significant in the study group. However, IL-8 and tumor necrosis factor alpha (TNF-alpha) levels did not change in either group (P>0.05). The amount of effusion decreased significantly in the study group (P=0.001) but not in the control group (P=0.133). It can be concluded that hyaluronan considerably decreased IL-6 levels, which correlated with clinical improvement, but had no effect on IL-8 and TNF-alpha levels in synovial fluid. However, larger studies with longer follow-up periods are needed to explain the effect of hyaluronan on cytokines.

Topics: Adjuvants, Immunologic; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hyaluronic Acid; Interleukin-6; Interleukin-8; Male; Middle Aged; Osteoarthritis, Knee; Pain Measurement; Range of Motion, Articular; Severity of Illness Index; Single-Blind Method; Synovial Fluid; Treatment Outcome; Tumor Necrosis Factor-alpha

Platelet-rich plasma (PRP) has been used extensively in clinical practice to treat patients with symptomatic knee osteoarthritis (OA). Leukocyte-poor PRP (LP-PRP) has been clinically preferred over leukocyte-rich PRP (LR-PRP); however, it is unclear which cytokine mediators of pain and inflammation are present in LR-PRP and LP-PRP from patients with mild to moderate knee OA in order to rationalize a specific formulation.. LP-PRP would be predominantly anti-inflammatory and have reduced nociceptive pain mediators compared with LR-PRP from the same individual with mild to moderate knee OA.. Controlled laboratory study.. A total of 24 unique samples of PRP were prepared in order to assess 48 samples of LR-PRP and LP-PRP taken from 12 patients (6 male and 6 female) with symptomatic knee OA of Kellgren-Lawrence grade 2 to 3. Patients underwent blood collection for LR-PRP and LP-PRP preparation through a double-spin protocol to obtain baseline whole blood, platelet concentration, and white blood cell subtypes. LR-PRP and LP-PRP from the same patient were produced at the same time and underwent a comprehensive panel through Luminex (multicytokine profiling) to assess key mediators of inflammation: interleukin 1 receptor antagonist (IL-1Ra), interleukin 4, 6, 8, and 10 (IL-4, IL-6, IL-8, and IL-10), IL-1β, tissue necrosis factor α (TNF-α), and matrix metalloproteinase 9 (MMP-9). To assess mediators of nociceptive pain, nerve growth factor (NGF) and tartrate resistant acid phosphatase 5 (TRAP5) were also assessed.. LR-PRP from patients with mild to moderate knee OA expressed significantly more IL-1Ra, IL-4, IL-8, and MMP-9 compared with LP-PRP formulations from the same patients. No significant differences were found between LR-PRP and LP-PRP in mediators of nociceptive pain-namely, NGF and TRAP5. Other mediators including TNF-α, IL-1β, IL-6, and IL-10 were also found to have no significant expression differences between LR-PRP and LP-PRP.. LR-PRP expressed significantly more IL-1Ra, IL-4, and IL-8, suggesting that LR-PRP may be more anti-inflammatory than LP-PRP. MMP-9 was expressed in higher concentrations in LR-PRP, suggesting that LR-PRP may be more chondrotoxic than LP-PRP.. LR-PRP was found to have a robust expression of anti-inflammatory mediators compared with LP-PRP and may be beneficial to patients with long-term knee OA where chronic low-grade inflammation is present. Mechanistic clinical trials are needed to elucidate the key mediators in both LR-PRP and LP-PRP to assess their effect on long-term progression of knee OA.

Topics: Anti-Inflammatory Agents; Female; Humans; Inflammation; Interleukin 1 Receptor Antagonist Protein; Interleukin-10; Interleukin-4; Interleukin-6; Interleukin-8; Leukocytes; Male; Matrix Metalloproteinase 9; Nerve Growth Factor; Osteoarthritis, Knee; Platelet-Rich Plasma; Prospective Studies; Treatment Outcome; Tumor Necrosis Factor-alpha

Despite the presence of chondrogenic progenitor cells (CPCs) in knee osteoarthritis patients they are unable to repair the damaged cartilage. This study aimed to evaluate the oxidative stress, cellular senescence, and senescence-associated secretory phenotype (SASP) in the CPCs derived from osteoarthritic cartilage and compare with the CPCs of healthy articular cartilage.. Isolated CPCs were characterized based on phenotypic expression of stem cell markers, clonogenicity, and tri-lineage differentiation assay. Production of ROS was measured using DCFDA assay. Cellular senescence in CPCs was assessed by senescence-associated beta-galactosidase assay and expression of senescence markers at the gene level using real-time PCR. Morphological features associated with senescent OA-CPCs were studied using scanning electron microscopy. To study SASP, the production of inflammatory cytokines was assessed in the culture supernatant using a flow-cytometer based cytometric bead array.. OA-CPCs exhibited elevated ROS levels along with a relatively high percentage of senescent cells compared to non-OA CPCs, and a positive correlation exists between ROS production and senescence. The morphological assessment of senescent CPCs revealed increased cell size and multiple nuclei in senescent OA-CPCs. These results were further validated by elevated expression of senescence genes p16, p21, and p53. Additionally, culture supernatant of senescent OA-CPCs expressed IL-6 and IL-8 cytokines indicative of SASP.. Despite exhibiting similar expression of stem cell markers and clonogenicity, CPCs undergo oxidative stress in diseased knee joint leading to increased production of intracellular ROS in chondrogenic progenitor cells that support cellular senescence. Further, senescence in OA-CPCs is mediated via the release of pro-inflammatory cytokines, IL-6 and IL-8.

Topics: Cartilage, Articular; Chondrocytes; Humans; Interleukin-6; Interleukin-8; Osteoarthritis, Knee; Senescence-Associated Secretory Phenotype; Stem Cells

There is a lack of in vitro models able to plausibly represent the inflammation microenvironment of knee osteoarthritis (OA). We analyzed the molecules released from OA tissues (synovial membrane, cartilage, infrapatellar fat pad) and investigated whether the stimulation of human synovial fibroblasts (SFs), with synthetic cytokines (IL-1β and TNF-α or IFN-γ) or conditioned media (CM) from OA tissues, influence the SFs' response, in the sense of pro-inflammatory cytokines, chemokines, growth factors, and degradative enzymes modulation. Human SFs were obtained from OA synovial membranes. SFs and their CM were analyzed for biomarkers, proliferation rate, protein profile and gene expression, before and after stimulation. Real-time PCR and multiplex assays quantified OA-related gene expression and biomolecule production. Unlike other activators, CM from OA synovial membrane (CM-SM), significantly up-regulated all genes of interest (IL-6, IL-8, MMP-1, MMP-3, RANTES, MCP-1, TSG-6, YKL-40) in SFs. Multiplex immunoassay analysis showed that levels of OA-related cytokines (IL-6, IL-8, MCP 1, IL-1Ra), chemokine (RANTES) and growth factor (VEGF), produced by CM-SM stimulated SFs, increased significantly compared to non-stimulated SFs. Molecules released from the SM from OA patients induces OA-like changes in vitro, in specific OA synovial populations (SFs). These findings promote the use and establish a compelling in vitro model that simulates the versatility and complexity of the OA disease. This model, in the future, will allow us to study new cell therapies or test drugs by reducing or avoiding animal models.

Topics: Animals; Chemokine CCL5; Chemokines; Culture Media, Conditioned; Cytokines; Fibroblasts; Humans; Interleukin-6; Interleukin-8; Osteoarthritis, Knee; Synovial Membrane

The aim of this study was to analyze local and systematic inflammatory status in knee osteoarthritis (KOA), focusing on intra-articular and remote adipose tissue depots, and to explore its potential association with metabolic syndrome (MetS).. Patients (n = 27) with end-stage KOA were enrolled in the study and samples from infrapatellar fat pad (IFP), synovium, subcutaneous adipose tissue (SAT), synovial fluid (SF), and serum were collected. In homogenates from the tissues, mRNA expression of developmental endothelial locus-1 (DEL-1) was determined. Interleukin 6 (IL-6) and interleukin 8 (IL-8) were measured in tissues and SF and serum samples by enzyme-linked immunosorbent assay.. Fifteen patients fulfilled MetS criteria (w-MetS group) and 12 did not (non-MetS). In the entire population, IL-6 levels were significantly higher in IFP compared to synovium (median (interquartile range), 26.05 (26.16) vs. 15.75 (14.8) pg/mg of total protein, p = 0.043), but not to SAT (17.89 (17.9) pg/mg); IL-8 levels were significantly higher in IFP (17.3 (19.3) pg/mg) and SAT (24.2 (26) pg/mg) when compared to synovium (8.45 (6.17) pg/mg) (p = 0.029 and < 0.001, respectively). Significantly higher IL-6 concentrations in SF were detected in w-MetS patients compared to non-MetS (194.8 (299) vs. 64.1 (86.9) pg/ml, p = 0.027). Finally, DEL-1 mRNA expression was higher in IFP compared to synovium (eightfold, p = 0.019).. Our findings support the critical role of IFP in knee joint homeostasis and progression of KOA. Furthermore, in KOA patients w-MetS, SAT is thought to play an important role in intra-knee inflammation via secretion of soluble inflammatory mediators, such as IL-6.

Topics: Adipose Tissue; Humans; Inflammation; Interleukin-6; Interleukin-8; Metabolic Syndrome; Osteoarthritis, Knee; RNA, Messenger

The increase of individuals with Osteoarthritis (OA) has generated an increase in public spending in the treatments, which are still not that effective. So, the purpose of this study was to analyze and compare four types of interventions: platelet-rich plasma (PRP), adipose-derived stem cells (ADSCs), ADSCs + PRP and the standard surgical video arthroscopy (All groups passed through standard arthroscopy before intervention). The evaluation was performed by applying the questionnaires Western Ontario McMaster Universities, Short Form Health Survey 36 and Visual Analog Pain Scale, also by analyzing the synovial fluid (inflammatory cytokines, enzymatic, colorimetric and viscosity analysis), this evaluation happened in two moments: before the surgical procedures and after 6 months of the interventions and also was made a comparison to standard arthroscopy. The questionnaires results showed a greater improvement in the scores of the domains analyzed in the ADSCs + PRP group, followed by the ADSCs and PRP group. In the evaluation of inflammatory cytokines, there was a significant reduction in the cytokine IL-1b only in the ADSCs + PRP group (46%) and ADSCs (31%), of IL-6 in the ADSCs + PRP group (72%), of IL-8 in the ADSCs + PRP group (50%) and ADSCs (31%), and TNF in the ADSCs + PRP group (46%). There was also a significant increase in the amount of total proteins (79%) in the control group and polymorphonuclear cells (47%) in the ADSCs + PRP group. Taking all the results into account, we infer that therapies with ADSCs + PRP and only ADSCs are safe and effective over 6 months for the improvement of pain, functional capacity and joint inflammation in volunteers with OA. It is also considered that the use of ADSCs + PRP, particularly, is a promising alternative to help manage this disease, due to the better results presented among the four propose interventions.

Topics: Humans; Hyaluronic Acid; Inflammation; Injections, Intra-Articular; Interleukin-6; Interleukin-8; Osteoarthritis, Knee; Pain; Platelet-Rich Plasma; Stem Cells; Treatment Outcome

To evaluate the changes of cytokines and immune cells after Intra-articular hyaluronic acid(IAHA)injections in patients with knee osteoarthritis (KOA).. Sixteen patients were included in the study, with a total of 65 IAHA injections. The Numeric Rating Scale (NRS) and Lysholm scores were evaluated at each visit. The immune cells and 14 cytokines of synovial fluid were analyzed at each visit. The association between immune cells and cytokines were examined.. IL-6 and IL-8 were the most common cytokines in the synovial fluid of KOA patients. The synovial fluid was orchestrated by macrophages (69%) and Lymphocytes (18%). Neutrophils were less to count of the total cell population (< 2%). The cytokines decreased significantly after the first injection and then tended to be stable. Lymphocytes increased a lot, while Macrophages decreased in the early stage, then increased after multiple injections. The proposition of M1 decreased in the early stage, then increased after multiple injections, while M2 increased consistently. M1 and M2 were positively associated with IL-6 and IL-8.. The synovial fluid of KOA patients was orchestrated by macrophages (69%) and Lymphocytes (18%) and cytokines like IL-6 and IL-8. IAHA may play an anti-inflammatory functional role through the decreased production of IL-6 and IL-8 by macrophages through polarization. The results from this study partially revealed the effect of IAHA on cytokines and immune cells change in KOA patients, and therapies targeting pathogenic cytokines and immune cells might be used to attenuate the knee joint inflammation and release pain.. ChiCTR2100050133; date registered 17 August 2021.

Topics: Cytokines; Humans; Hyaluronic Acid; Injections, Intra-Articular; Interleukin-6; Interleukin-8; Osteoarthritis, Knee; Synovial Fluid

Several cytokines and adipokines are related to clinical severity and progression in knee osteoarthritis. The aim of this study was to evaluate the associations of IL-8 with clinical severity and with local and systemic adipokines and cytokines. This is a Cross-sectional study including 115 women with symptomatic primary knee osteoarthritis with ultrasound-confirmed joint effusion. Age, symptoms duration and body mass index were collected. Radiographic severity was evaluated according to Kellgren-Lawrence. Pain and disability were assessed by Lequesne and Knee injury and Osteoarthritis Outcome Score pain, symptoms and function scales. Three inflammatory markers and five adipokines were measured by ELISA in serum and synovial fluid. Partial correlation coefficient (PCC) and corresponding 95% confidence interval were used to evaluate association. Synovial fluid IL-8 was significantly associated with clinical severity scales. After controlling for potential confounders, associations measured by a Partial Correlation Coefficient (PCC) remained essentially unaltered for Lequesne (PCC = 0.237), KOOS pain (PCC = - 0.201) and KOOS symptoms (PCC = - 0.209), KOOS function (PCC = - 0.185), although the later did not reach statistical significance. Also in synovial fluid samples, associations were found between IL-8 and TNF (PCC = 0.334), IL6 (PCC = 0.461), osteopontin (PCC = 0.575), visfatin (PCC = 0.194) and resistin (PCC = 0.182), although significance was not achieved for the later after statistical control for confounders. None of these associations were detected in serum. In conclusion, IL-8 was associated with clinical severity, inflammatory markers and adipokines in synovial fluid, but not in blood. Although the reported associations are weak to moderate in magnitude, these findings reinforce the notion that local and not systemic inflammation is more relevant to clinical severity in knee OA women with joint effusion.

Topics: Aged; Disease Progression; Female; Humans; Inflammation; Interleukin-8; Knee Joint; Middle Aged; Osteoarthritis, Knee; Patient Acuity; Synovial Fluid

Surgical knee joint distraction (KJD) leads to clinical improvement in knee osteoarthritis (OA) and also apparent cartilage regeneration by magnetic resonance imaging. We investigated if alteration of the joint's mechanical environment during the 6 week period of KJD was associated with a molecular response in synovial fluid, and if any change was associated with clinical response.. 20 individuals undergoing KJD for symptomatic radiographic knee OA had SF sampled at baseline, midpoint and endpoint of distraction (6 weeks). SF supernatants were measured by immunoassay for 10 predefined mechanosensitive molecules identified in our previous pre-clinical studies. The composite Knee injury and OA Outcome Score-4 (KOOS. 13/20 (65%) were male with mean age 54°±°5yrs. All had Kellgren-Lawrence grade ≥2 knee OA. 6/10 analytes showed statistically significant change in SF over the 6 weeks distraction (activin A; TGFβ-1; MCP-1; IL-6; FGF-2; LTBP2), P < 0.05. Of these, all but activin A increased. Those achieving the minimum clinically important difference of 10 points for KOOS. Detectable, significant molecular changes are observed in SF following KJD, that are remarkably consistent between individuals. Preliminary findings appear to suggest that increases in some molecules are associated with clinically meaningful responses. Joint distraction may provide a potential opportunity in the future to define regenerative biomarker(s) and identify pathways that drive intrinsic cartilage repair.

Topics: Activins; Cell Adhesion Molecules; Chemokine CCL2; External Fixators; Female; Fibroblast Growth Factor 2; Humans; Interleukin-6; Interleukin-8; Latent TGF-beta Binding Proteins; Male; Matrix Metalloproteinase 3; Middle Aged; Orthopedic Procedures; Osteoarthritis, Knee; Synovial Fluid; Tissue Inhibitor of Metalloproteinase-1; Transforming Growth Factor beta1; Treatment Outcome

Osteoarthritis (OA) of the knee is a multifactorial degenerative disease typically defined as the 'wear and tear' of articular joint cartilage. However, recent studies suggest that OA is a disease arising from chronic low-grade inflammation. We conducted a study to investigate the relationship between chronic inflammatory mediators present in both the systemic peripheral blood system and localised inflammation in synovial fluid (SF) of OA and non-OA knees; and subsequently made direct comparative analyses to understand the mechanisms that may underpin the processes involved in OA.. 20-Plex proteins were quantified using Human Magnetic Luminex® assay (R&D Systems, USA) from plasma and SF of OA (n = 14) and non-OA (n = 14) patients. Ingenuity Pathway Analysis (IPA) software was used to predict the relationship and possible interaction of molecules pertaining to OA.. There were significant differences in plasma level for matrix metalloproteinase (MMP)-3, interleukin (IL)-27, IL-8, IL-4, tumour necrosis factor-alpha, MMP-1, IL-15, IL-21, IL-10, and IL-1 beta between the groups, as well as significant differences in SF level for IL-15, IL-8, vascular endothelial growth factor (VEGF), MMP-1, and IL-18. Our predictive OA model demonstrated that toll-like receptor (TLR) 2, macrophage migration inhibitory factor (MIF), TLR4 and IL-1 were the main regulators of IL-1B, IL-4, IL-8, IL-10, IL-15, IL-21, IL-27, MMP-1 and MMP-3 in the plasma system; whilst IL-1B, TLR4, IL-1, and basigin (BSG) were the regulators of IL-4, IL-8, IL-10, IL-15, IL-18, IL-21, IL-27, MMP-1, and MMP-3 in the SF system.. The elevated plasma IL-8 and SF IL-18 may be associated with the pathogenesis of OA via the activation of MMP-3.

Topics: Aged; Biomarkers; Cartilage, Articular; Female; Humans; Interleukin-18; Interleukin-8; Knee Joint; Male; Osteoarthritis, Knee; Plasma; Synovial Fluid

To evaluate the anti-inflammatory effects of clinically relevant naproxen sodium (Nx) concentrations on human monocyte-derived macrophages in a controlled in vitro system and human primary synovial fluid (SF) cells.. Using phorbol 12-myristate 13-acetate, THP-1 human monocytic cells were differentiated into mature monocyte-derived macrophages in vitro then treated with Nx pre- or post-activating an inflammatory response with lipopolysaccharide (LPS) and hyaluronan (HA) fragments (n = 8/group). Cell culture supernatants were assessed for NF-κB activity and prostaglandin E. Compared to placebo treatment of THP-1 cells, low dose Nx (corresponding 27.5-440 mg/L orally) added both pre- and post-activation with LPS/HA, significantly reduced NF-κB activity and PGE2: mean reduction to 73%, 61%, 17% and 10% of placebo, respectively. LPS/HA treatment of primary OA SF cells significantly increased the number of IL-1β producing primary monocytes and macrophages, and by 24 h the overall production of secreted cytokines (IL-1β, IL-6, IL8, and TNF-α). Low dose Nx reduced the percentage of IL-1β producing primary monocytes and macrophages.. LPS/HA induced inflammation of THP-1 monocytic and primary human SF cells. Low dose Nx both prevented and reduced inflammatory responses of a human monocytic cell line and reduced IL-1β production by primary human SF monocytes and macrophages.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cytokines; Dinoprostone; Flow Cytometry; Humans; Hyaluronic Acid; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Lipopolysaccharides; Macrophages; Monocytes; Naproxen; Neutrophils; NF-kappa B; Osteoarthritis, Knee; Synovial Fluid; T-Lymphocytes; THP-1 Cells; Tumor Necrosis Factor-alpha

To determine the role of inflammatory biomarkers at 2 years post-anterior cruciate ligament (ACL) injury to predict radiographic knee osteoarthritis (OA) and magnetic resonance imaging (MRI)-defined knee OA at 5 years postinjury, with a secondary aim of estimating the concordance of inflammatory biomarkers assessed by MRI and synovial fluid (SF) analysis.. We studied 113 patients with acute ACL injury. Knee scans using 1.5T MRIs were read for Hoffa- and effusion-synovitis. Biomarkers of inflammation that we assessed included interleukin-6 (IL-6), IL-8, IL-10, tumor necrosis factor, and interferon-ɣ in serum and SF, and IL-12p70 in serum. We defined the outcome as radiographic knee OA (ROA) or MRI-defined OA (MROA) at 5 years. The area under the receiver operating characteristic curve (AUC), sensitivity, and specificity were evaluated in models that included MRI features only (model 1), inflammation biomarkers only (serum [model 2a] or SF [model 2b]), both MRI features and serum biomarkers (model 3a), or both MRI features and SF (model 3b) biomarkers. Linear regression analysis was used to evaluate the association between MRI features and SF biomarkers.. At 5 years postinjury, ROA was present in 26% of the injured knees, and MROA was present in 32%. The AUCs for ROA in each model were 0.44 (95% confidence interval [95% CI] 0.42, 0.47) for model 1, 0.62 (95% CI 0.59, 0.65) for model 2a, 0.53 (95% CI 0.50, 0.56) for model 2b, 0.58 (95% CI 0.55, 0.61) for model 3a, and 0.50 (95% CI 0.46, 0.53) for model 3b. The AUCs for MROA in each model were 0.67 (95% CI 0.64, 0.70) for model 1, 0.49 (95% CI 0.47, 0.52) for model 2a, 0.56 (95% CI 0.52, 0.59) for model 2b, 0.65 (95% CI 0.61, 0.68) for model 3a, and 0.69 (95% CI 0.66, 0.72) for model 3b. The concordance between MRI and SF biomarkers was statistically significant only for effusion-synovitis and IL-8.. Neither MRI-detected inflammation nor selected SF/serum inflammation biomarkers at 2 years postinjury predicted ROA or MROA at 5 years postinjury. Concordance between MRI and SF inflammatory biomarkers was weak.

Topics: Adult; Anterior Cruciate Ligament Injuries; Area Under Curve; Biomarkers; Female; Humans; Inflammation; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-6; Interleukin-8; Linear Models; Magnetic Resonance Imaging; Male; Osteoarthritis, Knee; Prognosis; ROC Curve; Synovial Fluid; Tumor Necrosis Factor-alpha; Young Adult

The aim of this study was to identify the molecules and pathways involved in the cross-talk between meniscus and synovium that may play a critical role in osteoarthritis (OA) pathophysiology. Samples of synovium and meniscus were collected from patients with early and end-stage OA and cultured alone or cocultured. Cytokines, chemokines, metalloproteases, and their inhibitors were evaluated at the gene and protein levels. The extracellular matrix (ECM) changes were also investigated. In early OA cultures, higher levels of interleukin-6 (IL-6) and IL-8 messenger RNA were expressed by synovium and meniscus in coculture compared with meniscus cultured alone. RANTES release was significantly increased when the two tissues were cocultured compared with meniscus cultured alone. Increased levels of matrix metalloproteinase-3 (MMP-3) and MMP-10 proteins, as well as increased release of glycosaminoglycans and aggrecan CS846 epitope, were observed when synovium was cocultured with meniscus. In end-stage OA cultures, increased levels of IL-8 and monocyte chemoattractant protein-1 (MCP-1) proteins were released in cocultures compared with cultures of meniscus alone. Chemokine (C-C motif) ligand 21 (CCL21) protein release was higher in meniscus cultured alone and in coculture compared with synovium cultured alone. Increased levels of MMP-3 and 10 proteins were observed when tissues were cocultured compared with meniscus cultured alone. Aggrecan CS846 epitope release was increased in cocultures compared with cultures of either tissue cultured alone. Our study showed the production of inflammatory molecules by synovium and meniscus which could trigger inflammatory signals in early OA patients, and induce ECM loss in the progressive and final stages of OA pathology.

Topics: Aged; Aged, 80 and over; Aggrecans; Cells, Cultured; Chemokine CCL2; Chemokine CCL21; Chemokine CCL5; Coculture Techniques; Extracellular Matrix; Female; Glycosaminoglycans; Humans; Inflammation; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 10; Matrix Metalloproteinase 3; Meniscus; Middle Aged; Osteoarthritis, Knee; Synovial Membrane

The aim of this study was to investigate cross-sectional associations between serum levels of IL-8 and the above outcomes in patients with knee osteoarthritis (OA).. A total of 160 subjects with clinical knee OA were included. Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score and Lequesne index were used to assess the joint symptom. Magnetic resonance imaging was used to measure knee structural abnormalities including infrapatellar fat pad (IPFP) signal intensity alternation. Knee radiographic OA was assessed by radiography using the Kellgren-Lawrence (K-L) grading system. Enzyme-linked immunosorbent assay was used to measure the serum levels of IL-8 and cartilage or bone biomarkers.. In multivariable analyses, serum IL-8 was positively associated with WOMAC weight-bearing pain (β 2.85, P = 0.028), WOMAC physical dysfunction (β 12.71, P = 0.048), and Lequesne index (β 1.65, P = 0.015), and had positive associations with IPFP signal intensity alteration (OR 3.18, P = 0.011) and serum levels of N-telopeptide of type I collagen (NTXI), N-terminal procollagen III propeptide (PIIINP), matrix metalloproteinase (MMP)3, and MMP13 (β 0.24-1.44, all P < 0.05) in patients with clinical knee OA. Furthermore, there were positive associations between IL-8 and WOMAC score (β 22.49, P = 0.037), K-L grades (OR 3.88, P = 0.013), and IPFP signal intensity alteration (OR 3.20, P = 0.033) in patients with radiographic OA.. Serum levels of IL-8 were positively associated with increased knee symptoms, IPFP signal intensity alteration, and serum levels of bone and/or cartilage biomarkers, suggesting that IL-8 may have a role to play in knee OA.Key Point• This study systemically investigates the associations between serum IL 8 and knee symptoms, joint structures, and cartilage or bone biomarkers in patients with knee osteoarthritis, and some significant associations have been found, suggesting that IL 8 may have a role to play in knee OA.

Topics: Adult; Aged; Biomarkers; Female; Humans; Interleukin-8; Knee Joint; Magnetic Resonance Imaging; Male; Middle Aged; Osteoarthritis, Knee; Radiography

The aim was to assess the activation and association of the NF-κB system across synovial membrane (SM) and articular cartilage (AC) in patients with knee osteoarthritis (OA) and ascertain its potential effects on catabolic mediator expression in advanced OA. SM and AC were obtained from 40 OA patients undergoing total knee arthroplasty and from 19 postmortem control subjects. NF-κB subunit RelA in nuclear and cytosolic fractions and NF-κB1-DNA binding in nuclear extracts was assessed by ELISA, whereas

Topics: Adult; Aged; Cartilage, Articular; Cells, Cultured; Disease Progression; DNA; Female; Humans; Inflammation; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 3; Middle Aged; NF-kappa B p50 Subunit; Osteoarthritis, Knee; Protein Binding; Signal Transduction; Synovial Membrane; Transcription Factor RelA; Transcriptional Activation

Osteoarthritis (OA) is a major cause of morbidity and incapacity in the elderly. This study evaluates serum levels of the chemokines CCL2, CXCL8, CXCL9, and CXCL10 in 16 patients with primary OA of the knees, and investigates how treatment with hydroxychloroquine (HCQ) for 4 months affects these chemokine levels.. Thirteen elderly patients received a placebo. Healthy control groups consisted of 10 elderly individuals (age > 60 years) with no clinical or radiological evidence of OA (CT-O), and 10 young adult individuals, (CT-Y group, age < 40 years).. The CT-Y group presented lower levels of all chemokines studied, in comparison to the other groups. HCQ treatment did not alter the serum levels of CCL2 (P = 0.80), CXCL8 (P = 0.76), CXCL9 (P = 0.95) and CXCL10 (P = 0.74) in OA patients.. Hydroxychloroquine treatment did not alter the serum levels of CCL2, CXCL8, CXCL9 or CXCL10 in patients with OA of the knees, although increased serum levels correlated with aging for all subjects, including controls.

Topics: Age Factors; Aged; Aging; Antirheumatic Agents; Biomarkers; Case-Control Studies; Chemokine CCL2; Chemokine CXCL10; Chemokine CXCL9; Female; Humans; Hydroxychloroquine; Interleukin-8; Male; Middle Aged; Osteoarthritis, Knee; Time Factors; Treatment Outcome; Up-Regulation

Degenerative osteoarthritis (OA) has been associated with elevated synovial fluid cytokines. It is unclear whether traumatic knee injuries are a trigger to the chemical process that leads to OA. The purpose of this study was to compare the synovial fluid cytokine levels between knees undergoing arthroscopy due to a documented inciting injury and knees undergoing primary arthroplasty due to end-stage OA without a previous inciting injury. Synovial fluid samples were prospectively collected from knees undergoing arthroscopic surgeries due to ligamentous or meniscal knee injuries (knee injury group,

Topics: Adolescent; Adult; Aged; Arthroscopy; Cytokines; Female; Humans; Interleukin-6; Interleukin-8; Knee Injuries; Knee Joint; Male; Middle Aged; Osteoarthritis, Knee; Synovial Fluid; Tumor Necrosis Factor-alpha; Young Adult

Compared with subcutaneous adipose tissue (SCAT), infrapatellar fat pad (IFP), the main knee intra-articular adipose tissue (IAAT), has an inflammatory phenotype in patients with osteoarthritis (OA). We phenotyped suprapatellar fat pad (SPFP) and hip acetabular fat pad (AFP), two other IAATs, to determinate the unique signature of IAATs compared with SCAT.. IFP, SPFP, AFP and autologous SCAT were obtained from patients with OA during total knee (n=38) or hip replacement (n=5). Fibrosis and adipocyte area were analysed by histology and vascularisation, leucocyte and mast cell infiltration were analysed by immunohistochemistry for von Willebrand factor, leucocytes and tryptase, respectively. Secretion of interleukin (IL)-6, IL-8 and prostaglandin E. Fibrosis, vascularisation and leucocyte and mast cell infiltration were greater in IAATs than SCAT, and levels of IL-6, IL-8 and PGE. IFP but also SPFP and AFP (gathered under the term 'IAAT') may play a deleterious role in OA by affecting joint homeostasis because of their inflammatory phenotype and their close interaction with synovium in the same functional unit.

Topics: Adipocytes; Adipose Tissue; Adolescent; Adult; Aged; CD36 Antigens; Culture Media, Conditioned; Dinoprostone; Fatty Acid-Binding Proteins; Female; Gene Expression; Hip Joint; Homeodomain Proteins; Humans; Interleukin-6; Interleukin-8; Knee Joint; Lipase; Lipoprotein Lipase; Male; Membrane Proteins; Middle Aged; Osteoarthritis, Hip; Osteoarthritis, Knee; Phenotype; PPAR gamma; RNA, Messenger; Subcutaneous Fat; Synoviocytes; Young Adult

Norepinephrine (NE) was measured in synovial fluid of trauma patients and sympathetic nerve fibers were detected in healthy and osteoarthritic (OA) joint tissues indicating that cartilage pathophysiology might be influenced by sympathetic neurotransmitters. The aim of this study was to elucidate the mostly unknown role of NE in OA chondrocyte metabolism and inflammatory responses.. Articular cartilage was received after total knee replacement surgery from OA patients. Expression of adrenergic receptors (AR) and tyrosine hydroxylase (TH) was tested with end point polymerase chain reaction (PCR) and immunohistochemistry. 3-dimensional (3D) cell cultures were employed to analyze effects of NE on chondrocyte cell metabolism and the expression of interleukins (ILs), matrix metalloproteases (MMPs), tissue inhibitor of metalloproteases (TIMPs), glycosaminoglycan (GAG) and collagen II under non- and inflammatory conditions. Chondrocyte monolayer cultures were used to specify AR subtypes, to analyze cell cycle distribution and to determine catecholamines in cell culture supernatants.. AR subtypes and TH were detected in chondrocytes, whereas NE was not released in measurable amounts. 10(-6) M NE reversed IL-1β induced changes in IL-8, MMP-13, GAG and collagen II expression/production indicating for β-AR signaling. Additionally, NE caused cell cycle slow down and decreased proliferation via β-AR signaling. 10(-8) M NE increased the number of proliferating cells and induced apoptosis via α1-AR signaling.. NE affects chondrocytes from OA cartilage regarding inflammatory response and its cell metabolism in a dose dependent manner. The sympathetic nervous system (SNS) may have a dual function in OA pathology with preserving a stable chondrocyte phenotype via β-AR signaling and OA pathogenesis accelerating effects via α-AR signaling.

Topics: Adrenergic alpha-Agonists; Aged; Aged, 80 and over; Cartilage, Articular; Cell Culture Techniques; Chondrocytes; Collagen Type II; Female; Glycosaminoglycans; Humans; Immunohistochemistry; Inflammation; Interleukin-1beta; Interleukin-8; Interleukins; Knee Joint; Male; Matrix Metalloproteinase 13; Matrix Metalloproteinases; Middle Aged; Norepinephrine; Osteoarthritis, Knee; Polymerase Chain Reaction; Receptors, Adrenergic; Receptors, Adrenergic, alpha; Receptors, Adrenergic, beta; Tissue Inhibitor of Metalloproteinases; Tyrosine 3-Monooxygenase

To explore the ability of chemokines in plasma to detect the presence of pre-X-rays defined knee degeneration and the extent (burden).. A total of 181 subjects (75 control subjects, 47 pre-X-KD patients and 50 X-KOA patients) were included and subdivided into three subgroups. Articular cartilage loss in pre-X-KD patients were scored on the basis of the ICRS classification during the arthroscopy or documented on MRI with chondral WORMS. The severity of X-KOA was graded using the Kellgren-Lawrence classification through the posterior-anterior knee X-rays. The concentrations of the inflammatory cytokines and chemokines in plasma were quantified using Luminex microbead-based suspension array (SA) and were cross-validated by enzyme-linked immunosorbent assay (ELISA).. CCL3 in plasma showed the highest ability to discriminate pre-X-KD patients from the controls with an AUC of 0.799. At a cutoff value of 0.168 pg/ml, the sensitivity was 70.21%, the specificity was 96.00%, the positive predictive value was 91.67% and the negative predictive value was 83.72%. As to define disease burden, the plasma levels of resistin, IL6, IL8, CCL3 and CCL4 showed significant association with the severity of X-rays defined knee OA, with regard to the KL classification. Moreover, significant elevation of IL6, IL8, CCL3 and CCL4 levels in plasma were observed in severe knee OA patients (KL grade IV) compared with those with pre-X-KD (KL grade 0-I).. We firstly showed that the plasma CCL3 could be potential serum biomarker for knee OA with the capacity to detect pre-X-rays defined changes and stage the severity of damage in knee.

Topics: Adult; Biomarkers; Case-Control Studies; Chemokine CCL3; Chemokine CCL4; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Osteoarthritis, Knee; Predictive Value of Tests; Radiography; Resistin; Sensitivity and Specificity; Severity of Illness Index

We previously identified an association between bone sialoprotein (BSP) and osteoarthritic (OA) chondrocyte hypertrophy but the precise role of BSP in ostearthritis (OA) has not been extensively studied. This study aimed to confirm the association between BSP and OA chondrocyte hypertrophy, to define its effect on molecules produced by chondrocytes and to analyse its association with cartilage degradation and vascular density at the osteochondral junction.. Human OA chondrocytes were cultivated in order to increase hypertrophic differentiation. The effect of parathyroid hormone-related peptide (PTHrP), interleukin (IL)-1β or tumour necrosis factor (TNF)-α on BSP was analysed by real-time reverse transcription polymerase chain reaction (RT-PCR) and western blot. The effects of BSP on OA chondrocytes production of inflammatory response mediators (IL-6, nitric oxide), major matrix molecule (aggrecan), matrix metalloprotease-3 and angiogenic factors (vascular endothelial growth factor, basic fibroblast growth factor, IL-8, and thrombospondin-1) were investigated. BSP was detected by immunohistochemistry and was associated with cartilage lesions severity and vascular density.. PTHrP significantly decreased BSP, confirming its association with chondrocyte hypertrophy. In presence of IL-1β, BSP stimulated IL-8 synthesis, a pro-angiogenic cytokine but decreased the production of TSP-1, an angiogenesis inhibitor. The presence of BSP-immunoreactive chondrocytes in cartilage was associated with the severity of histological cartilage lesions and with vascular density at the osteochondral junction.. This study supports the implication of BSP in the pathology of OA and suggests that it could be a key mediator of the hypertrophic chondrocytes-induced angiogenesis. To control chondrocyte hypertrophic differentiation is promising in the treatment of OA.

Topics: Aggrecans; Blotting, Western; Chondrocytes; Fibroblast Growth Factor 2; Humans; In Vitro Techniques; Integrin-Binding Sialoprotein; Interleukin-1beta; Interleukin-6; Interleukin-8; Matrix Metalloproteinase 3; Nitric Oxide; Osteoarthritis, Knee; Parathyroid Hormone-Related Protein; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Thrombospondin 1; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factors

Fibromyalgia and osteoarthritis may present a relationship with the concentration of cytokines. The aim of this study was to compare the serum concentrations of IL-12p70, tumor necrosis factor, IL-10, IL-6, IL-1β, and IL-8 in patients with knee osteoarthritis and fibromyalgia.. The study included 53 women (71.2±7.6 years old) diagnosed with knee osteoarthritis with moderate-to-severe pain (visual analog scale >4) for at least 3 months. Sixty women (54.1±8.1 years old) diagnosed with fibromyalgia according to the American College of Rheumatology criteria and with moderate-to-severe pain (visual analog scale >4) also participated in this study. For the dosage of cytokines, blood was collected in the morning: 5 mL from the cubital vein. The material was centrifuged at 4°C, separated into 100 μL aliquots and stored at -80°C until processing. Serum concentrations of the studied cytokines were assessed using the BD Cytometric Bead Array method. Data were analyzed with Student's t-test and the Mann-Whitney U test.. We found higher levels of IL-6, IL-10, and IL-1β in fibromyalgia patients. After adjustment of age as a covariate, there was no statistically significant difference in the concentration of any cytokine between fibromyalgia and knee osteoarthritis patients.. Patients with knee osteoarthritis and fibromyalgia with the same duration and intensity of pain demonstrate similar concentrations of cytokines. Aging may play a role in cytokine profile, a finding not so extensively addressed in the literature and one that should be further investigated.

Topics: Aged; Cytokines; Female; Fibromyalgia; Humans; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Middle Aged; Osteoarthritis, Knee; Pain Measurement; Tumor Necrosis Factor-alpha

Inflammation of the synovial membrane (synovitis) is considered to drive the process that leads to osteoarthritis. However, the relationships between the mediators of inflammation and the properties of wear particles are not fully understood. In this study, the levels of IL-6 and IL-8 were assessed in different grades of osteoarthritis to determine whether their concentrations in the synovial fluid correlate with specific characteristics of wear particles. This study has found that the size, adhesion and nano-surface roughness of wear particles have medium strong to strong correlations with IL-6 and IL-8. This study provided evidence that the characteristics of wear particles contain valuable information for grading the disease process and the need for further evaluation of the association of properties of wear particles and the inflammatory process.

Topics: Female; Foreign-Body Reaction; Humans; Interleukin-6; Interleukin-8; Knee Joint; Knee Prosthesis; Male; Middle Aged; Nanoparticles; Osteoarthritis, Knee; Particle Size

Osteoarthritis (OA) is a chronic musculoskeletal disease characterized by progressive destruction of articular cartilage, OA lead to chronic pain and functional restrictions in affected joints. The present study was to investigate the role of osteopontin (OPN) in the athogenesis of OA through studying the effect of OPN on expression of IL-6 and IL-8 inflammatory factors in human OA chondrocytes.. One-step type II collagenase digestive method was used to isolate OA chondrocytes from sectional cartilage specimens of 16 primary knee OA patients received total knee replacement surgery. Synchronized first-generation chondrocytes were then treated with OPN (100 ng/ml or 1 µg/mL). The changes in cell morphology of OA chondrocytes were analyzed before and after treated with OPN; and the expression levels of IL-6 and IL-8 were evaluated by real-time q-PCR.. Chondrocytes were successfully isolated from human OA knee cartilage, and the viability of isolated chondrocytes was 92.11±3.13%. Adherent chondrocytes formed clusters of irregular polygonal shape with intercellular pseudopodia extension. After OPN treatment, cells became fusiform or irregularly shaped, and the number of intercellular pseudopodia decreased significantly. The mRNA expression of IL-6 increased to 1.83 times at 0.1 µg/ml of OPN and 3.1 times at the dose of 1 µg/ml; the expression of IL-8 increased to 1.57 and 3.27 times at the dose of 0.1 µg/ml and 1 µg/ml respectively.. OPN could up-regulate expression of IL-6 and IL-8 cytokines in human OA chondrocytes, and the expression increased with the increasing concentration of OPN, which might be one of the potential mechanisms of OPN in the development of OA.

Topics: Adult; Aged; Cells, Cultured; Chondrocytes; Female; Gene Expression Regulation; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Osteoarthritis, Knee; Osteopontin; Treatment Outcome

Connective tissue growth factor (CTGF), also known as CCN2, is a key proinflammatory mediator. In the present study, the involvement of the CTGF signaling pathway in human knee osteoarthritis (OA) fibroblast-like synoviocytes (FLSs) was investigated. FLSs were isolated from human OA synovium and incubated with CTGF in the absence or presence of interleukin‑1β (IL‑1β). The expression of relevant genes and proteins was analyzed by qPCR, western blotting and enzyme-linked immunosorbent assay (ELISA). Matrix metalloproteinase (MMP) activity and nuclear factor (NF)-κB activation were also evaluated. CTGF stimulation resulted in the significant production of IL-6, IL-8, C-C motif ligand 2 (CCL2), CCL20, MMP-1 and MMP-3 in FLSs in the presence, but not in the absence, of IL-1β. CTGF also enhanced the levels of phosphorylated extracellular signal-related kinase 1/2 (ERK1/2) and p38. In addition, CTGF at 25 ng/ml, in the presence of IL‑1β, significantly potentiated NF-κB activation. The results indicated that CTGF interacted with IL‑1β in FLSs to promote the inflammatory response in the synovium, leading to the initiation of the inflammatory cascade. These results support the proinflammatory role of CTGF in synovitis and joint destruction in OA.

Topics: Aged; Cells, Cultured; Chemokine CCL2; Chemokine CCL20; Connective Tissue Growth Factor; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; NF-kappa B; Osteoarthritis, Knee; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Synovial Membrane

Human six-transmembrane epithelial antigen of prostate4 (STEAP4), an ortholog of mouse tumor necrosis factor-α-induced adipose-related protein (TIARP), plays a role in tumor necrosis factor (TNF)-dependent arthritis models. However, its role in rheumatoid arthritis (RA) is still obscure. This study explored such a role for STEAP4. The expressions of STEAP4, TNFα, and IL-6 were compared in synovia of RA and osteoarthritis patients. STEAP4 induction was examined in TNFα-stimulated fibroblast-like synoviocytes (FLS) in vitro. FLS (with/without TNFα stimulation) were also analyzed for IL-6 expression after STEAP4 knockdown, using siRNA or transfection with STEAP4-plasmid DNA. IL-8, cell proliferation, and apoptosis were also evaluated in STEAP4-overexpressing FLS. The expression of STEAP4 in joints correlated with TNFα expression, specifically in RA synovium. In the cultured FLS, STEAP4 protein expression was augmented by TNFα activation, and localized in endosomal/lysosomal compartments. STEAP4 downregulation by siRNA enhanced the expression of IL-6 mRNA, while STEAP4 overexpression suppressed IL-6 and IL-8 expression, inhibited cell proliferation, and induced apoptosis via caspase-3. The results indicated that human STEAP4 is regulated by TNFα in synovium, where it controls IL-6 secretion and proliferation of FLS, suggesting that STEAP4 might potentially suppress the pathogenesis of TNFα-induced arthritis such as RA.

Topics: Arthritis, Rheumatoid; Cell Proliferation; Fibroblasts; Gene Expression; Gene Silencing; Humans; Interleukin-6; Interleukin-8; Jurkat Cells; Membrane Proteins; Osteoarthritis, Knee; Oxidoreductases; RNA, Small Interfering; Synovial Membrane; Transfection; Tumor Necrosis Factor-alpha

IL-8 and its murine equivalent keratinocyte chemoattractant (Kc), chemokines produced by chondrocytes, contribute to the pathophysiology of osteoarthritis. However, the mechanisms leading to their production are poorly known. We aimed to investigate whether mechanical (compression), inflammatory (IL-1β) and metabolic (visfatin) stresses may induce the release of Kc when applied on murine cartilage.. Mouse cartilage explants were subjected to intermittent compression for 4, 6 and 24h. Primary cultures of immature murine articular chondrocytes were obtained by enzymatic digestion of articular cartilage from 6-days-old newborns mice. The effect of compression, IL-1β (10, 50, 100pg/mL) and of visfatin (5μg/mL) on the release of Kc was assessed by ELISA. IL-8 levels in conditioned media from human OA joint tissues (cartilage or synovium) were also assessed.. In comparison with non-compressed explants, loading increased Kc release of 3.2-, 1.9- and 2.0-fold at 4, 6 and 24h respectively (P<0.004, n=9). IL-1β triggered an increase of Kc release by primary cultured chondrocytes of 4.1-, 15.5- and 35.2-fold at 10, 50 and 100pg/mL of IL-1β respectively (P<0.05, n=4). Likewise, visfatin (5μg/mL) induced an increase in Kc release of 56.5±25.2 fold (P=0.002, n=6). IL-8 was released in conditioned media by synovium as well as by cartilage.. We show for the first time that IL-8/Kc is highly responsive to mechanical, inflammatory and metabolic stresses, strengthening the hypothesis that IL-8/Kc could be added to the cytokines which may have a deleterious impact in osteoarthritis.

Topics: Animals; Animals, Newborn; Cartilage, Articular; Cells, Cultured; Chemokine CXCL1; Chondrocytes; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Humans; Interleukin-1beta; Interleukin-8; Mice; Nicotinamide Phosphoribosyltransferase; Osteoarthritis, Knee; Receptors, Interleukin-8B; Stress, Mechanical

Chemokines proved able to induce release of enzymes relevant in cartilage damage. The present study addressed the levels of CXCL8 and CCL5 and the potential role of these chemokines in predicting the morphological changes in the course of osteoarthritis (OA). Synovial fluid (SF) and blood serum were obtained from 20 patients undergoing knee replacement surgery because of OA. For comparison, samples were also obtained from another 20 patients during diagnostic or therapeutic arthroscopy performed because of knee injury. The samples were analyzed for CXCL8 and CCL5 using enzyme-linked immunosorbent assay. SF from the group with OA showed significantly (p = 0.024) increased levels of CXCL8 when compared with the group after knee injury. We have not demonstrated any significant correlation between chemokine expression and clinical or radiological signs of OA. Mediators of inflammation are the potential predicting factors of OA, however, with respect to examined chemokines development of a diagnostic test can be limited by the low serum concentration and lack of correlation with clinical and radiological signs of the disease.

Topics: Adult; Aged; Arthroplasty, Replacement, Knee; Biomarkers; Case-Control Studies; Chemokine CCL5; Enzyme-Linked Immunosorbent Assay; Humans; Inflammation Mediators; Interleukin-8; Middle Aged; Osteoarthritis, Knee; Radiography; Synovial Fluid; Young Adult

Environmental factors are involved in RA pathogenesis and epidemiological studies have suggested that smoking is an environmental risk factor for RA. The 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is one of the major toxic components in cigarettes. To clarify the biological effects of smoking in RA, we investigated the role of TCDD in RA pathogenesis.. Human synovial tissue was obtained from RA and OA patients and aryl hydrocarbon receptor (AhR) expression in these tissues was evaluated using immunohistochemistry and real-time PCR. Expression of various cytokines was measured by real-time PCR following stimulation of RA synoviocytes with different concentrations of TCDD. To study the role of AhR, we treated RA synoviocytes with alpha-naphthoflavone, a known AhR antagonist. To evaluate which signal transduction pathways were stimulated by the TCDD-AhR interaction, we used inhibitors of nuclear factor-kappaB (NF-kappaB) and extra-cellular stimulus-activated kinase (ERK).. Higher AhR mRNA and protein levels were observed in RA synovial tissue than in OA tissue. TCDD up-regulated the expression of IL-1beta, IL-6 and IL-8 through binding to AhR, and this effect was transmitted via the NF-kappaB and ERK signalling cascades. AhR expression in synovial cells was up-regulated by TNF-alpha.. TNF-alpha activates AhR expression in RA synovial tissue, and that cigarette smoking and exposure to TCDD enhances RA inflammatory processes. TCDD induces inflammatory cytokines via its association with AhR, resulting in stimulation of the NF-kappaB and ERK signalling cascades. Thus TCDD exposure, such as smoking exacerbates RA pathophysiology.

Topics: Arthritis, Rheumatoid; Cells, Cultured; Dose-Response Relationship, Drug; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; NF-kappa B; Osteoarthritis, Knee; Polychlorinated Dibenzodioxins; Receptors, Aryl Hydrocarbon; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Synovial Membrane; Tumor Necrosis Factor-alpha; Up-Regulation

Fibroblast-like synoviocytes (FLS) are potentially directly involved in the propagation of inflammation. We have previously shown evidence of an expanded activated population of natural killer (NK) cells in spondylarthritis (SpA) patients. In the present study, we sought to determine whether the interaction between NK cells and FLS from SpA patients results in a proinflammatory response.. Autologous NK cells and FLS were obtained from 6 patients with SpA, 4 patients with rheumatoid arthritis (RA), and 8 patients with osteoarthritis (OA). Physical interactions between NK cells and FLS were studied by time-lapse phase-contrast microscopy. Fluorescence-activated cell sorting was used to study the activation, proliferation, and survival of NK cells in contact with FLS. Cytokine and stromal factor production were measured by a multiple cytokine bead assay.. NK cells both adhered to and migrated beneath the FLS monolayer (pseudoemperipolesis). FLS from SpA and RA patients supported increased pseudoemperipolesis, activation, cytokine production, and survival of NK cells. The production of proinflammatory cytokines, including interleukin-6 (IL-6), IL-8, IL-1beta, and IL-15, was increased in cocultures of NK cells and FLS, particularly in those from RA and SpA patients. Production of interferon-gamma, RANTES, and matrix metalloproteinase 3 (MMP-3) by NK cell and FLS coculture was greatest in SpA patients. Surface expression of IL-15 on FLS was significantly increased in SpA and RA patients, but not OA patients. Blockade with an IL-15 monoclonal antibody resulted in increased apoptosis of NK cells.. FLS promote the migration, activation, and survival of NK cells. The interaction of NK cells with FLS results in increased IL-15 expression by FLS and the production of proinflammatory chemokines, cytokines, and MMPs, which may contribute to joint inflammation. This response was much more marked in SpA and RA patients as compared with OA patients.

Topics: Adult; Aged; Arthritis, Rheumatoid; Cell Communication; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Cytokines; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interferon-gamma; Interleukin-15; Interleukin-1beta; Interleukin-6; Interleukin-8; Killer Cells, Natural; Male; Matrix Metalloproteinase 3; Middle Aged; Osteoarthritis, Knee; Spondylarthritis; Synovial Membrane

This study was performed to investigate whether synovial proliferation (SP) differentially affects hypoxia in the joint cavities of rheumatoid arthritis (RA) and osteoarthritis (OA) patients. Thirty RA and 42 OA patients who underwent synovitis assessment were classified into two groups based on the presence or absence of SP, as revealed by musculoskeletal ultrasonography. Synovial fluids (SFs) from the knee joints were analyzed for interleukin (IL)-8, pO(2), and white blood cell counts and blood samples were analyzed for erythrocyte sedimentation rate (ESR). No difference was found between the OA patients with and without SP in terms of SF oxygen tension (SF pO(2)) or IL-8 level, whereas the RA patients had significantly lower SF pO(2) levels in their knee joints than did the OA patients with SP, and the RA patients had higher levels of IL-8 in their joints than did the OA patients. The counts of infiltrated immune cells in the SF and tissues were much higher for patients with RA and SP than for those with OA and SP. The ESRs were not found to be correlated with SP in OA patients but were negatively correlated with SF pO(2) levels in RA patients. We conclude that ultrasonographically detected SP in OA patients does not generate a more hypoxic SF than that found in OA patients without SP. The SFs from RA patients with SP are hypoxic, which indicates that SP may have different impacts on hypoxia in the joint cavities of RA and OA patients.

Topics: Arthritis, Rheumatoid; Blood Sedimentation; Cell Proliferation; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hypoxia; Immunohistochemistry; Interleukin-8; Knee Joint; Male; Osteoarthritis, Knee; Oxygen; Severity of Illness Index; Synovial Fluid; Synovial Membrane; Ultrasonography

To investigate the relationship between macrophage migration inhibitory factor (MIF) levels and clinical measures in rheumatoid arthritis (RA), and the potential for regulation of angiogenesis in RA.. Serum and synovial fluid (SF) levels of MIF and vascular endothelial growth factor (VEGF) in patients with RA were determined by sandwich ELISA, and the relationships among MIF, VEGF, and RA clinical measures were analyzed. RA synovial fibroblasts were cultured with recombinant human MIF (rhMIF) and the production of VEGF and interleukin 8 (IL-8) were measured in the conditioned media. The angiogenic effect of MIF was examined using established measures of angiogenesis in vitro.. Erythrocyte sedimentation rate, C-reactive protein, and the daily dosage of oral prednisolone were correlated with SF levels of MIF. The SF levels of MIF were found to be higher in patients with bony erosion than in those without (69.2 +/- 11.4 ng/ml vs 44.0 +/- 6.2 ng/ml; p = 0.045). MIF levels had good correlation with VEGF levels (r = 0.52, p < 0.001 in sera, and r = 0.6, p < 0.001 in SF). Production of the angiogenic factors VEGF and IL-8 was enhanced in cultured RA synovial fibroblasts stimulated by rhMIF. Endothelial tube formation was augmented when the endothelial cells were cultured with the conditioned media from rhMIF-pretreated SF mononuclear cells, and this phenomenon was reversed by anti-VEGF antibody.. SF MIF may reflect the clinical activity in patients with RA, and rhMIF induces the angiogenic factors in RA synovial fibroblasts. These results suggest that MIF may be an important cytokine in the perpetuation of the angiogenic and inflammatory processes in patients with RA.

Topics: Angiogenesis Inducing Agents; Arthritis, Rheumatoid; Biomarkers; Cells, Cultured; Culture Media, Conditioned; Dose-Response Relationship, Drug; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Female; Fibroblasts; Humans; Interleukin-8; Leukocytes, Mononuclear; Macrophage Migration-Inhibitory Factors; Male; Middle Aged; Neovascularization, Physiologic; Osteoarthritis, Knee; Recombinant Proteins; Synovial Fluid; Synovial Membrane; Up-Regulation; Vascular Endothelial Growth Factor A

Activated synoviocytes play important roles in the progression of human osteoarthritis (OA). Intra-articular injection of high molecular weight hyaluronic acid (HMW-HA) has been used as viscosupplementation for knee OA but its effect on synoviocytes remains undisclosed. This study aims to investigate the effects of HMW-HA on the gene expression of 16 OA-associated cytokines and enzymes, including interleukin (IL)-1beta, IL-6, IL-8, leukemia inhibitory factor (LIF), tumor necrosis factor (TNF)-alpha, TNF-alpha converting enzyme (TACE), matrix metalloproteinase (MMP)-1, MMP-2, MMP-3, MMP-9, MMP-13, tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2, aggrecanase-1, aggrecanase-2, and inducible nitric oxide synthase (iNOS), in fibroblast-like synoviocytes (FLS) from patients with early stage OA.. Synovial fluid-derived FLS were obtained from the knees of 15 patients with early stage OA. IL-1-stimulated or unstimulated FLS were cultured with or without the treatment of 600-800kDa HMW-HA. Moreover, blocking experiments with anti-CD44 monoclonal antibodies (mAb) were used to examine the involvement of CD44 in HMW-HA effects. We designed and validated the real-time quantitative polymerase chain reaction (Q-PCR) assays with SYBR Green dyes for simultaneous quantification of the expression of the 16 genes.. HMW-HA down-regulated IL-8 and iNOS gene expression in unstimulated FLS and down-regulated aggrecanase-2 and TNF-alpha gene expression in IL-1-stimulated FLS. CD44 blocking inhibited the down-regulatory effects of HMW-HA on gene expression.. HMW-HA may have a structure-modifying effect for OA by down-regulation of aggrecanase-2 in FLS. HMW-HA also has an anti-inflammatory effect by down-regulation of TNF-alpha, IL-8, and iNOS in FLS. These effects may be mediated through the interaction of CD44 and HMW-HA.

Topics: ADAM Proteins; ADAMTS5 Protein; Cytokines; Dose-Response Relationship, Drug; Down-Regulation; Fibroblasts; Humans; Hyaluronan Receptors; Hyaluronic Acid; Interleukin-1; Interleukin-8; Molecular Weight; Nitric Oxide Synthase Type II; Osteoarthritis, Knee; Polymerase Chain Reaction; Synovial Fluid; Tumor Necrosis Factor-alpha

The aim of the present study was to determine synovial levels of ELR (+) CXC chemokines, known to attract mainly neutrophils to inflamed tissues by binding the neutrophil chemokine receptors CXCR1 and CXCR2 and promoting neovascularization in patients with various inflammatory disorders. The study group consisted of 14 patients with Behçet's disease and nine with familial Mediterranean fever. Fourteen patients with rheumatoid arthritis and 16 with osteoarthritis served as controls. Synovial chemokine levels were measured by two-step sandwich enzyme-linked immunosorbent assay, and significant differences were found in the various chemokines studied. In addition to its angiogenic properties, increased synovial levels of interleukin-8 by attraction of more neutrophils to synovial fluids might also be responsible for the acute synovitis in patients with Behçet's disease. However, the absence of chronic changes with the eventual development of pannus and erosions might result from relatively lower expression of interleukin-8 and the transient, short-lived nature of the arthritis observed in these patients.

Topics: Adult; Angiogenesis Inhibitors; Arthritis, Rheumatoid; Behcet Syndrome; Chemokine CXCL1; Chemokine CXCL5; Chemokines, CXC; Familial Mediterranean Fever; Female; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Male; Microtubule-Associated Proteins; Middle Aged; Neovascularization, Pathologic; Osteoarthritis, Knee; Synovial Fluid

To obtain fibroblast-like synovial cells (FLS) from synovial fluid (SF).. SF aspirated from joints of patients with rheumatoid arthritis (RA), other types of inflammatory arthritis, and osteoarthritis (OA) was centrifuged and the resulting cell pellet resuspended in growth medium. After 2 days, nonadherent cells were removed. FLS were also cultured from surgical specimens of synovial tissue (td-FLS). Phenotype characterization of fluid derived FLS (fd-FLS) was accomplished by flow cytometry and immunohistochemistry staining. Tumor necrosis factor-alpha (TNF-alpha) induced interleukin 6 (IL-6), IL-8, and cyclooxygenase 2 (COX-2) mRNA levels were assessed.. Second and later passage fd-FLS exhibited uniform fibroblast-like morphology. Fd-FLS and td-FLS expressed a similar profile of cell surface antigens including the fibroblast marker Thy-1. Less than 2% of either cell type expressed surface markers characteristic of dendritic cells, phagocytic cells, T cells, or leukocytes. Immunohistochemistry staining revealed the presence of fibroblast products prolyl-4 hydroxylase, procollagen I, and procollagen III in both culture types. TNF-a induced increases in IL-6, IL-8, and COX-2 mRNA were suppressed by dexamethasone in both fd-FLS and td-FLS.. FLS can be cultured from SF. The fibroblast phenotype was confirmed by analysis of surface antigens and intracellular proteins. Inflammatory mediators produced after stimulation of both fd-FLS and td-FLS were suppressed by dexamethasone. In addition to providing a more accessible source of FLS, fd-FLS may also facilitate study of synovial cells in early RA when tissue specimens are not readily available.

Topics: Arthritis; Arthritis, Rheumatoid; Cells, Cultured; Cyclooxygenase 2; Dexamethasone; Fibroblasts; Flow Cytometry; Gene Expression; Humans; Immunoenzyme Techniques; Interleukin-6; Interleukin-8; Knee Joint; Membrane Proteins; Osteoarthritis, Knee; Phenotype; Prostaglandin-Endoperoxide Synthases; RNA, Messenger; Synovial Fluid; Synovial Membrane; Tumor Necrosis Factor-alpha

Interleukin-8 (IL-8) plays an important role in the migration of inflammatory cells into the synovium and joint fluids in rheumatoid arthritis (RA). This study was undertaken to investigate the IL-8 inductive activity of the macrophage migration inhibitory factor (MIF) in RA synovial fibroblasts. The regulatory mechanism of IL-8 was compared with that of IL-1beta.. MIF-induced IL-8 and IL-1beta transcriptional activation was studied in RA synovial fibroblasts by Northern blot analysis, enzyme-linked immunosorbent assay, and electromobility shift assay. The effect of anti-MIF antibody administration on murine passive collagen-induced arthritis (CIA) was also evaluated by histologic examination and reverse transcriptase-polymerase chain reaction.. MIF up-regulated the IL-8 messenger RNA (mRNA) and protein levels in a dose-dependent manner. The IL-8 mRNA up-regulation started 1 hour poststimulation by MIF, and reached a maximum level at 6 hours. IL-1beta mRNA was also up-regulated by MIF. The mRNA up-regulation of IL-8 and IL-1beta by MIF was inhibited by 2 tyrosine kinase inhibitors, a protein kinase C (PKC) inhibitor, an activator protein 1 (AP-1) inhibitor, and by an NF-kappaB inhibitor. A cAMP-dependent kinase inhibitor did not inhibit it. MIF enhanced AP-1 and NF-kappaB binding activities in a dose-dependent manner. Passive CIA enhanced mRNA levels of macrophage inflammatory protein 2 and cytokine-induced neutrophil chemoattractants and, moreover, migration and proliferation of inflammatory cells within the synovium, which were suppressed by administration of an anti-MIF antibody.. MIF may play an important role in the migration of inflammatory cells into the synovium of rheumatoid joints via induction of IL-8. MIF up-regulates IL-8 and IL-1beta mRNA via tyrosine kinase-, PKC-, AP-1-, and NF-kappaB-dependent pathways.

Topics: Arthritis, Rheumatoid; Chemokine CXCL2; Chemokines; Cytokines; Dose-Response Relationship, Drug; Enzyme Inhibitors; Fibroblasts; Gene Expression; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Macrophage Migration-Inhibitory Factors; Monokines; NF-kappa B; Osteoarthritis, Knee; Protein Kinase Inhibitors; RNA, Messenger; Sialoglycoproteins; Signal Transduction; Synovial Membrane; Transcription Factor AP-1; Transcription, Genetic; Up-Regulation