In vitro lead cation-hyaluronic acid interaction

It is the purpose of this investigation to study the in vitro spontaneous interaction lead-hyaluronic acid using the equilibrium dialysis technique. The following working conditions were used: membrane retention capacity starting from 12000 D; solvent was an aqueous buffer solution (pH 7.4); relation of internal-external volume of 0.5 : 25 mL: dialysis time 24 hours; lead concentration in dialyzing solution 4 micrograms/mL (always outside the bag); temperature 4 degrees C; concentrations of hyaluronic acid 0.3, 1.2, 2.1 and 3.0 mg/mL; concentration of plasma proteins 1.5 mg/mL; lead was determined in the internal solution of the bag, and was measured quantitatively by atomic absorption spectrometry. Testing separately hyaluronic acid and plasma proteins both types of marcromolecules turned out to bind lead. As the concentration of hyaluronic acid increases, the amount of lead retained increases as well, although this relation is only approximately linear. At the normal concentrations of plasma proteins and hyaluronic acid in synovial fluid, the proteins solution bound more lead than hyaluronic acid solution 16.2 +/- 2.1 and 5.7 +/- 0.4 micrograms/mL respectively (P < 0.0001). What these data show ist that both, hyaluronic acid and proteins are involved in the in vitro lead retention. The effect of both ligands must be synergistic, because when the two molecules are present, a higher amount of lead is retained compared to the amount bound separately, suggesting that lead can replace calcium by complexation with mucopolysaccharides; this explains the lead accumulation in synovial fluid since this fluid is rich in hyaluronic acid and proteins.