Abstract
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.
Highlights
Synovial fluid (SF) is a viscoelastic system with a characteristic non-Newtonian behaviour, consisting of hyaluronan (HA) and proteins, at least partly involved in a loose physical 3D-network based on secondary interactions of electrostatic nature [1]
We have recently studied the influence of fibrinogen as a model of protein on the rheology of HA, and demonstrated the physical interaction at neutral pH which should be based on electrostatic interactions [1]
This work was performed to investigate the role of pre-treatments of synovial fluids (SF) on its rheological properties and composition determined by steric exclusion chromatography with triple detection (SEC)
Summary
Synovial fluid (SF) is a viscoelastic system with a characteristic non-Newtonian behaviour, consisting of hyaluronan (HA) and proteins, at least partly involved in a loose physical 3D-network based on secondary interactions of electrostatic nature [1]. HA is claimed to have a molecular weight (MW) in the range of 4 to 5 × 106 with a concentration ranging from 2.5 to 4 mg/mL [12,13,14,15,16] This means a total amount of HA ranging from 4 to 8 mg per human knee joint, since the volume of SF is assumed to be 1 to 2 mL in normal conditions [12,13]. HA is an anionic polyelectrolyte, able to interact with positively charged groups of proteins even at a very low level (it was estimated as 2% protein bound to HA) [8]
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