Role of Different Pre-Treatments on Composition and Rheology of Synovial Fluids

Polymers (Impact Factor: 3.68). 12/2009; 1(1). DOI: 10.3390/polym1010016
Source: DOAJ


Different pre-treatments applied to synovial fluids (SF) before their analyses are tested to characterize SF after storage under different conditions and to investigate their evolution along a viscosupplementation treatment. The main techniques proposed involve steric exclusion chromatography with triple detection (SEC) and viscometry; it is the first time that such a study is developed. SEC gives the molecular weight distribution and concentration of hyaluronan (HA) and proteins separately; the steady state viscosity is always non-Newtonian and not directly related to SF composition. Pre-treatment of SF (storage in cold, filtration, centrifugation) allows us to conclude that, in order to store SF, it is best to freeze it, even if in some cases, viscosity is modified but not the composition. All the data obtained (including protease pre-treatment) allow us to conclude that a small fraction of HA-protein complex forms a loose 3D-network and controls the rheology.

Download full-text


Available from: Pierre Mathieu,
28 Reads
  • Source
    • "Important components of SF are lubricin and some proteins from blood plasma (γ-globulin and albumin), which enhance the lubricating properties of SF (Oates, 2006). The importance of HA and proteins for the lubricating properties of SF was also described (Swann et al., 1985; Rinaudo et al., 2009). "
    Biomaterials - Physics and Chemistry, 11/2011; , ISBN: 978-953-307-418-4
  • [Show abstract] [Hide abstract]
    ABSTRACT: Purpose Osteoarthritis (OA) is a degenerative joint disease that is characterized by the breakdown of articular cartilage. Rheology of synovial fluid is of interest because of its significance in joint lubrication. However, there are still many questions related to synovial fluid rheology and its relation with OA. Although viscosupplementation has been used as a treatment for OA for many years, its clinical effect remains controversial. Therefore, the purpose of this study is to determine in vitro the effect of different viscosupplements on the rheological properties of synovial fluid. Methods A detailed rheological characterization of synovial fluid from 26 patients undergoing total knee arthroplasty (TKA) was performed. The rheological properties of the osteoarthritic synovial fluid added with viscosupplements were compared with those of the healthy human synovial fluid in order to assess the restorative effect on naturally occurring synovial fluid within the joint. Furthermore, the stability of rheological properties of synovial fluid mixed with viscosupplements in vitro over 8 weeks was analyzed. Results The addition of a cross-linked viscosupplement to OA synovial fluid was found to be more efficient than linear ones in improving the overall rheological properties of synovial fluid. The findings indicate that the rheological properties of SF mixed with VSs changed over 8 weeks. Conclusions The findings from this study will lead to a better understanding of the role of viscosupplements in joint lubrication and OA.
    Biomedical Engineering Letters 11/2011; 1(4). DOI:10.1007/s13534-011-0034-7
  • [Show abstract] [Hide abstract]
    ABSTRACT: Synovial fluid (SF) is a difficult biological matrix to analyse due to its complex non-Newtonian nature. This can result in poor assay repeatability and potentially inefficient use of precious samples. This study assessed the impact of SF treatment by hyaluronidase and/or dilution on intra-assay precision using the Luminex and Meso Scale Discovery (MSD) multiplex platforms. SF was obtained from patients with knee osteoarthritis at the time of joint replacement surgery. Aliquots derived from the same sample were left untreated (neat), 2-fold diluted, 4-fold diluted or treated with 2mg/ml testicular hyaluronidase (with 2-fold dilution). Preparation methods were compared in a polysterene-bead Luminex 10-plex (N=16), magnetic-bead Luminex singleplex (N=7) and MSD 4-plex (N=7). Each method was assessed for coefficient of variation (CV) of replicate measurements, number of bead events (for Luminex assays) and dilution-adjusted analyte concentration. Percentage recovery was calculated for dilutions and HAse treatment. Hyaluronidase treatment significantly increased the number of wells with satisfactory bead events/region (95%) compared to neat (48%, p<0.001) in the polystyrene-bead Luminex assay, but the magnetic-bead Luminex assay achieved ≥50 bead events irrespective of treatment method. Hyaluronidase treatment resulted in lower intra-assay CVs for detectable ligands (group average CV<10%) than neat, 2-fold and 4-fold dilution (CV~25% for all, p<0.05) in both polystyrene- and magnetic-bead Luminex assays. In addition, measured sample concentrations were higher and recovery was poor (elevated) after hyaluronidase treatment. In the MSD 4-plex, within-group comparison of the intra-assay CV or concentration was not conclusively influenced by SF preparation. However, only hyaluronidase treatment resulted in CV<25% for all samples for TNF-α. There was no effect on analyte concentrations or recovery. Hyaluronidase treatment can improve intra-assay precision and assay signal of SF analysis by multiplex immunoassays and should be recommended for SF biomarker research, particularly using the Luminex platform.
    Journal of immunological methods 08/2012; 386(1-2):22-30. DOI:10.1016/j.jim.2012.08.012 · 1.82 Impact Factor
Show more