UBC Theses and Dissertations
Flow conductivity of solutions of hyaluronic acid : effects of concentration and molecular weight Lam, Luk Sang
Hyaluronic acid plays an important role in regulating the transport of fluid and solutes in the interstitium. The concentration and molecular weight of hyaluronic acid in different connective tissues are different. These factors influence the hydraulic flow conductivity, K', of connective tissues. An experimental study of the effect of concentration and molecular weight of hyaluronic acid on the hydraulic flow conductivity is the subject of this work. Hyaluronic acid of different molecular weights were obtained by fractionating commercially available hyaluronic acid using ion-exchange column chromatography. The results were not reproducible, partly because of the elution process was not continuous. Nevertheless, three molecular weight fractions (6.99 to 11.1 X 10⁵ ) were obtained. Hyaluronic acid of lower molecular weights (0.454 to 1.65 X 10⁵) were obtained by acid hydrolysing some of the chromatographed fractions for 15 min., 1 hour and 2 hours. A more homogeneous hyaluronic acid fraction (M.W. = 1.96 X10⁵) was obtained by fractionating hyaluronic acid materials acid hydrolysed for 15 min. The hydraulic flow conductivity of solutions of hyaluronic acid can be calculated from the sedimentation coefficient of the solutions at 20°C, S₂₀‚ measured by ultracentrifugation. Centrifugation experiments determining the S₂₀ of the molecular weight fractions of hyaluronic acid at various concentrations were therefore undertaken. The results showed that S₂₀ decreased with increased concentration of hyaluronic acid. Also, the curves of as a function of hyaluronic acid concentration, c, converged at high concentration, indicating that a three dimensional molecular network is formed at high concentration and the extent of entanglement between molecules is the same for the high and low M.W. fractions. At lower concentrations, for the acid hydrolysed fractions, S₂₀ increased with M.W., which is in agreement with past sedimentation data. For the non-acid hydrolysed fractions, the difference in S₂₀ between two higher M.W. fractions is small, and the lowest M.W. fraction has consistently higher S₂₀ than the higher M.W. fractions. This finding does not agree with past literature results, and the difference in results is most probably due to experimental errors. However, when the fractionated non-acid hydrolysed fractions are taken as a high M.W. group (M.W. = 6.99 to 11.1X10⁵) and the acid hydrolysed fractions as a low M.W. group (M.W. = 0.454 to 1.96X10⁵), the curves of S₂₀ as a function of c of the low M.W. group fall below those of the high M.W. group, which is in agreement with past sedimentation data. The hydraulic conductivities (K'), calculated from S₂₀ data, for all the HA fractions varied inversely with concentration. The log-log plots of K' versus c compared well with the results of Ethier (1986). The K' versus c relationships for all the fractions converged at high concentrations. At low concentrations, the HA molecules of the high M.W. group has a higher K' than those of the low M.W. group.
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