Saline-Assisted Aspirations for Collecting Synovial Fluid From Noneffused Knees: Technique and Validation

2014 ◽  
Vol 23 (4) ◽  
Author(s):  
Jeffrey B. Driban ◽  
Nicole Cattano ◽  
Easwaran Balasubramanian ◽  
Michael R. Sitler ◽  
Mamta Amin ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Cohort Studies ◽  
Interleukin 1 ◽  
Biochemical Changes ◽  
Science Laboratory ◽  
Secondary Analyses ◽  
Joint Trauma ◽  
Fluid Biomarker ◽  
Radiographic Knee Osteoarthritis ◽  
Synovial Fluid Protein

Context: To better understand why a knee develops osteoarthritis after joint trauma we need to assess the local biochemical changes. Unfortunately, it is challenging to obtain synovial fluid from a knee with no effusion. Objective: To describe the authors' protocol for aspirating synovial fluid from noneffused knees. Second, they demonstrate the validity of this method by evaluating the relationships between normalized and raw biomarker concentrations among knees with effusion (undergoing a traditional aspiration) and without effusion (requiring a saline-assisted aspiration). Design: Validation study based on secondary analyses from 2 cohort studies. Setting: Outpatient orthopedic clinic and basic-science laboratory. Participants: Participants had moderate to severe radiographic knee osteoarthritis (n = 15 with and 11 without effusion) and no osteoarthritis or effusion (n = 4). Interventions: The same orthopedic surgeon performed all synovial-fluid joint aspirations, including saline-assisted aspirations. Main Outcome Measures: The authors used multiplex enzyme-linked immunosorbent assays to determine 7 synovial-fluid biomarker concentrations. They then calculated correlations between raw and normalized (to total synovial-fluid protein content) biomarker concentrations. Results: The authors excluded 1 sample collected with a saline-assisted aspiration because it contained blood. Normalized biomarker concentrations had positive associations with raw biomarker concentrations (r = .77-99), with the exception of interleukin-13 and interleukin-1Β among knees that underwent a saline-assisted aspiration. Excluding interleukin-1Β, associations between normalized and raw biomarker concentrations were consistent between knees that had a saline-assisted or traditional aspiration. Conclusions:Saline-assisted aspiration is a valid technique for assessing the local biochemical changes in knees without effusion.

There is an Association of Synovial Interleukin-6 Levels With Chondral Damage in Anterior Cruciate Ligament–Deficient Knees

2021 ◽  
pp. 155633162199200
Author(s):  
Ravi Gupta ◽  
Anil Kapoor ◽  
Sourabh Khatri ◽  
Dinesh Sandal ◽  
Gladson David Masih
Keyword(s):  
Anterior Cruciate Ligament ◽  
Synovial Fluid ◽  
Acl Reconstruction ◽  
Inflammatory Cytokines ◽  
Interleukin 6 ◽  
Cruciate Ligament ◽  
Interleukin 1 ◽  
Chondral Damage ◽  
Anterior Cruciate ◽  
Acl Deficient

Background: Osteoarthritis (OA) in the anterior cruciate ligament (ACL)–deficient knee is seen in approximately 50% of affected patients. Possible causes include biochemical or biomechanical changes. Purpose: We sought to study the correlation between inflammatory cytokines and chondral damage in ACL-deficient knees. Methods: Seventy-six male patients who underwent ACL reconstruction were enrolled in a cross-sectional study. Synovial fluid was aspirated before surgery and analyzed for levels of the inflammatory cytokines tumor necrosis factor-α, interleukin-1 (IL-1), and interleukin-6 (IL-6). At the time of ACL reconstruction, the severity of chondral damage was documented as described by the Outerbridge classification. Results: Patients with grade 2 or higher chondral damage were observed to have elevated IL-6 levels when compared to patients who had no chondral damage. Interleukin-6 levels had no correlation with the duration of injury. Conclusion: Elevated levels of IL-6 in synovial fluid were associated with chondral damage in ACL-deficient knees. Further study is warranted to determine whether inflammatory cytokines contribute to the development of OA of the knee after ACL injury.

scholarly journals AB0108 IRAK4 INHIBITION SUPPRESSES PROINFLAMMATORY CYTOKINE PRODUCTION FROM HUMAN MACROPHAGES STIMULATED WITH SYNOVIAL FLUID FROM RHEUMATOID ARTHRITIS PATIENTS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1353.2-1353
Author(s):  
A. Yadon ◽  
D. Ruelas ◽  
G. Min-Oo ◽  
J. Taylor ◽  
M. R. Warr
Keyword(s):  
Rheumatoid Arthritis ◽  
Synovial Fluid ◽  
Signaling Pathways ◽  
Proinflammatory Cytokines ◽  
Proinflammatory Cytokine ◽  
Cytokine Production ◽  
Interleukin 1 ◽  
Proinflammatory Cytokine Production ◽  
Human Macrophages ◽  
Percent Suppression

Background:Rheumatoid arthritis (RA) is characterized by chronic, uncontrolled joint inflammation and tissue destruction. Macrophages are thought to be key mediators in both the initiation and perpetuation of this pathology.1,2The RA synovium contains a complex inflammatory milieu that can stimulate macrophage-dependent production of proinflammatory cytokines through multiple signaling pathways.1,2Existing evidence indicates that toll-like receptors (TLRs) and interleukin-1 receptors (IL-1R) along with their agonists, damage-associated molecular patterns (DAMPs) and IL-1β, are highly expressed in RA joints and are important mediators of synovial macrophage activation and proinflammatory cytokine production.1-9IRAK4 (interleukin-1 receptor-associated kinase 4) is a serine/threonine kinase that facilitates TLR and IL-1R signaling in many cell types, including macrophages.10IRAK4 inhibition represents an opportunity to reduce proinflammatory cytokine production in the joints of patients with RA.Objectives:To investigate the effect of a highly selective IRAK4 inhibitor on proinflammatory cytokine production from human macrophages stimulated with synovial fluid from patients with RA.Methods:Primary human monocytes from 2 independent donors were differentiated for 6 days with granulocyte-macrophage colony-stimulating factor (GM-CSF) to generate human monocyte-derived macrophages (hMDMs). hMDMs were then pretreated with an IRAK4 inhibitor for 1 hour and subsequently stimulated for 24 hours with RA synovial fluid from 5 patients. Culture supernatants were then assessed for secretion of proinflammatory cytokines by MesoScale Discovery.Results:RA synovial fluid stimulation of hMDMs resulted in the production of several proinflammatory cytokines, including IL-6, IL-8, and TNFα. Pretreatment of hMDMs with an IRAK4 inhibitor resulted in the dose-dependent inhibition of IL-6, IL-8, and TNFα production, with an average EC50± SD of 27 ± 31, 26 ± 41, and 28 ± 22 nM, respectively. Maximal percent suppression ± SD of IL-6, IL-8, and TNFα were 76 ± 8.8, 73 ± 15, and 77 ± 13, respectively. To evaluate the specific IRAK4-dependent signaling pathways mediating this response, hMDMs were pretreated with inhibitors of TLR4 (TAK242) and IL-1R (IL-1RA) prior to stimulation with RA synovial fluid. Both TAK242 and IL-1RA inhibited proinflammatory cytokine production. For TAK242, maximal percent suppression ± SD of IL-6, IL-8, and TNFα were 39 ± 25, 48 ± 24, and 50 ± 21, respectively. For IL-1RA maximal percent suppression ± SD of IL-6, IL-8, and TNFα were 18 ± 18, 20 ± 23, and 16 ± 18, respectively. The broad range of inhibition across each stimulation highlights the complexity and variability in the signaling pathways mediating proinflammatory cytokine production from hMDMs stimulated with RA synovial fluid, but demonstrates that RA synovial fluid can stimulate proinflammatory cytokine production in hMDMs, at least partly, through IRAK4-dependent pathways.Conclusion:This work demonstrates that IRAK4 inhibition can suppress proinflammatory cytokine production from macrophages stimulated with synovial fluid from patients with RA and supports a potential pathophysiological role for IRAK4 in perpetuating chronic inflammation in RA.References:[1]Smolen JS, et al.Nat Rev Dis Primers.2018;4:18001.[2]Udalova IA, et al.Nat Rev Rheumatol.2016;12(8):472-485.[3]Joosten LAB, et al.Nat Rev Rheumatol.2016;12(6):344-357.[4]Huang QQ, Pope RM.Curr Rheumatol Rep.2009;11(5):357-364.[5]Roh JS, Sohn DH.Immune Netw.2018;18(4):e27.[6]Sacre SM, et al.Am J Pathol.2007;170(2):518-525.[7]Ultaigh SNA, et al.Arthritis Res Ther.2011;13(1):R33.[8]Bottini N, Firestein GS.Nat Rev Rheumatol.2013;9(1):24-33.[9]Firestein GS, McInnes IB.Immunity.2017;46(2):183-196.[10]Janssens S, Beyaert R.Mol Cell.2003;11(2):293-302.Disclosure of Interests:Adam Yadon Employee of: Gilead, Debbie Ruelas Employee of: Gilead, Gundula Min-Oo Employee of: Gilead, James Taylor Employee of: Gilead, Matthew R. Warr Employee of: Gilead

Recombinant human interleukin-1 β–induced increase in levels of proteoglycans, stromelysin, and leukocytes in rabbit synovial fluid

1992 ◽  
Vol 35 (7) ◽  
pp. 799-805 ◽  
Author(s):  
Joseph Mcdonnell ◽  
Lori A. Hoerrner ◽  
Michael W. Lark ◽  
Coral Harper ◽  
Tanvi Dey ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Interleukin 1

scholarly journals An Alternative Macrophage Activation Pathway Regulator, CHIT1, May Provide a Serum and Synovial Fluid Biomarker of Periprosthetic Osteolysis

2017 ◽  
Vol 14 (2) ◽  
pp. 148-152 ◽  
Author(s):  
Samir K. Trehan ◽  
Lester Zambrana ◽  
Jonathan E. Jo ◽  
Ed Purdue ◽  
Athanos Karamitros ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Macrophage Activation ◽  
Periprosthetic Osteolysis ◽  
Activation Pathway ◽  
Fluid Biomarker

Equine synovial fluid protein equalization via combinatorial peptide ligand libraries

2021 ◽  
Vol 1 (4) ◽  
pp. 18-22
Author(s):  
Pablo Fueyo ◽  
Marco Galleguillos ◽  
Cristóbal Dörner ◽  
Pedro A. Smith ◽  
Francisca Godoy ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Sodium Dodecyl ◽  
Electrophoretic Pattern ◽  
Peptide Ligand ◽  
Joint Pathology ◽  
Coomassie Blue ◽  
Standard Procedures ◽  
Sds Page ◽  
New Biomarkers ◽  
Synovial Fluid Protein

To gain further knowledge of the equine synovial fluid (SF) proteome, we propose a protocol based on the equalization of the relative concentrations of its proteins, which leads to the modification of the standard electrophoretic pattern revealing low-abundance proteins that otherwise remain undetected. Fresh SF samples were collected from ten macroscopically normal metacarpophalangeal joints of crossbred horses. The samples were processed using standard procedures as the control and via combinatorial peptide ligand libraries (CPLL) using low ionic forces (NaH2PO4 10 mM) at different pHs (4.0, 7.0, and 9.3) with 10% sodium dodecyl sulfate (SDS) and 25 mM DTT for protein resolubilization. Proteins were then separated by conventional 8% SDS-PAGE and stained with coomassie blue. After separation of the equalized proteins, there was a significant reduction in the albumin (the most abundant protein in the SF) and, at the same time, other protein bands arise that were not visible without CPLL processing. In addition, there was variation in the protein profiles at different pHs. The results suggest that protein equalization of the equine SF by CPLL could be a useful tool to better understand the articular homeostasis and/or for the detection of new biomarkers of joint pathology.

Occurrence of interleukin-1 in human synovial fluid: detection by RIA, bioassay and presence of bioassay-inhibiting factors

1989 ◽  
Vol 9 (2) ◽  
pp. 53-58 ◽  
Author(s):  
J. B. Smith ◽  
M. H. Bocchieri ◽  
L. Sherbin-Allen ◽  
M. Borofsky ◽  
J. L. Abruzzo
Keyword(s):  
Synovial Fluid ◽  
Interleukin 1 ◽  
Inhibiting Factors ◽  
Fluid Detection ◽  
Human Synovial Fluid

scholarly journals Synovial Fluid Biomarker Concentrations in Knees with Symptomatic Meniscus Injury Compared to Asymptomatic Contralateral Knees

2019 ◽  
Vol 7 (7_suppl5) ◽  
pp. 2325967119S0035
Author(s):  
Andrew Clair ◽  
Matthew T. Kingery ◽  
Utkarsh Anil ◽  
Lena Kenny ◽  
Eric Jason Strauss
Keyword(s):  
Synovial Fluid ◽  
Cartilage Damage ◽  
Arthroscopic Surgery ◽  
Low Grade ◽  
Current Analysis ◽  
Duration Of Symptoms ◽  
Cartilage Lesions ◽  
Meniscus Injury ◽  
Symptomatic Knee ◽  
Fluid Biomarker

Objectives: Changes in the joint microenvironment following an intra-articular injury have been implicated in the pathogenesis of knee osteoarthritis. Few studies have evaluated alterations in the joint microenvironment in the setting of meniscus injury. The purpose of the current study was to determine the changes in synovial fluid biomarker concentrations caused by meniscus pathology by comparing samples from injured, symptomatic knees to samples from asymptomatic contralateral knees. Methods: Patients undergoing surgery for unilateral meniscus injury were prospectively enrolled in this institutional review board approved study from October 2011 to December 2016. A cohort was formed consisting of patients that had synovial fluid samples collected from both the injured and contralateral uninjured knee at the time of arthroscopic surgery. Patients with ligamentous injury of the knee were excluded from the current analysis. Synovial fluid samples were collected just prior to incision and the concentrations of 10 biomarkers of interest were determined using a multiplex magnetic bread immunoassay. Results: The current analysis included synovial fluid samples from 82 knees (41 operative and 41 contralateral knees) from 41 patients undergoing arthroscopic surgery to treat a symptomatic meniscus injury. The mean age of patients was 49.86 +/- 11.75 years. Based on linear mixed effects models, there were significantly greater concentrations of 4 of the 5 pro-inflammatory biomarkers in symptomatic knees compared to asymptomatic knees when controlling for the duration of symptoms, BMI, age, and the random effects of by-patient variability. Knees with symptomatic meniscus injuries had 126.8 times greater concentration of IL-6, 2.7 times greater concentration of MCP-1, 2.0 times greater concentration of MIP-1beta, and 5.4 times greater concentration of MMP-3 compared to the contralateral, asymptomatic knee (Table 1). When controlling for the chronicity of the injury, presence of synovitis, and age of the patient, knees with concomitant high-grade cartilage lesions (ICRS 3 or 4) were associated with 2.1 times greater concentration of MCP-1, 1.9 times greater concentration of MIP-1beta, and 3.4 times greater concentration of VEGF compared to knees with concomitant low-grade cartilage lesions (ICRS 1 or 2). When controlling for the other variables, the presence of synovitis was associated with an 89.5% lower concentration of TIMP-1 compared to operative knees without synovitis. The age of the patient was found to affect the concentrations of IL-6, MCP-1, and VEGF. For all knees included in the study, each 1 year increase in age was associated with a 6% increase in IL-6, 3% increase in MCP-1, and 4% increase in VEGF (Figure 1). Conclusion: This study is the first that examines the synovial fluid biomarker concentrations in the setting of a symptomatic isolated meniscus injury. We demonstrated that 4 of the 5 proinflammatory biomarkers that were tested are found in greater concentration in the symptomatic knee. Furthermore, we described the effects of associated cartilage damage, synovitis, and patient age on biomarker concentrations. Understanding the implication of these alterations in the intra-articular microenvironment in the setting of meniscal pathology may hold the key to identifying treatment targets in an effort to prevent the onset of post-meniscectomy osteoarthritis. [Table: see text]

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