Biophysical Journal #2004/050302 (revised) Hindered Convection of Macromolecules in Hydrogels

Hindered convection of macromolecules in gels was studied by measuring the sieving coefficient (Q) of narrow fractions of Ficoll (Stokes-Einstein radius, r s = 2.7-5.9 nm) in agarose and agarose-dextran membranes, along with the Darcy permeability (k). To provide a wide range of k, varying amounts of dextran (volume fractions £ 0.011) were covalently attached to agarose gels with volume fractions of 0.040 or 0.080. As expected, Q decreased with increasing r s or with increasing concentrations of either agarose or dextran. For each molecular size, Q plotted as a function of k fell on a single curve for all gel compositions studied. The dependence of Q on k and r s was predicted well by a hydrodynamic theory based on flow normal to the axes of equally spaced, parallel fibers. Values of the convective hindrance factor (K c , the ratio of solute to fluid velocity), calculated from Q and previous equilibrium partitioning data, were unexpectedly large; although K c £ 1.1 in the fiber theory, its apparent value ranged generally from 1.5 to 3. This seemingly anomalous result was explained on the basis of membrane heterogeneity. Convective hindrances in the synthetic gels were quite similar to those in glomerular basement membrane, when compared on the basis of similar solid volume fractions and values of k. Overall, the results suggest that convective hindrances can be predicted fairly well from a knowledge of k, even in synthetic or biological gels of complex composition.