The effects of pH and ionic strength on intrafibrillar hydration in articular cartilage.

The hydration of articular cartilage is an essential determinant of its load bearing capacity. Here we have examined the dependence of the amount of intrafibrillar water, associated with the collagen molecules in both native and PG-depleted cartilage specimens, on the pH and ionic strength of the bathing solution, in the presence and absence of an externally applied pressure. We found that high ionic strength reduces the collagen intermolecular spacing over a large pH range: this is consistent with the electrostatic nature of the interactions between the charged groups within the intrafibrillar space. We also found that as the pH is lowered from neutral to approximately 3, there is, as expected, a gradual increase in the overall positive charge of the intrafibrillar compartment. However, surprisingly, this is not accompanied by an increase in the intrafibrillar hydration; only at pH 1.8 does the amount of intrafibrillar water increase markedly. We suggest that, rather than overall intrafibrillar charge, it is specific local axial and azimuthal relationships among collagen molecules in the fibril, and more particularly, among their charged amino acid residues, that determine the intermolecular collagen spacing, and hence intrafibrillar hydration.