Hyaluronan interactions with hydroxyapatite do not alter in vitro hydroxyapatite crystal proliferation and growth

Abstract

The interaction of hyaluronan (Mr range 80-120 x 10(4)) with poorly crystalline hydroxyapatite, such as is found in calcified cartilage and bone, was studied to challenge the hypothesis that free hyaluronan found in proteoglycan aggregate preparations could affect in vitro mineralization. Using a Langmuir adsorption isotherm, based on uronic acid content, hyaluronan was found to bind to hydroxyapatite with an affinity K of 0.12 ml/microgram uronate and N = 6.8 micrograms uronate/m2 hydroxyapatite binding sites. This is contrasted with K = .047 ml/microgram uronate and N = 9.0 micrograms uronate/m2 for a bovine nasal proteoglycan monomer preparation. Although the proteoglycan monomer and aggregate preparations have been reported to inhibit hydroxyapatite growth at concentrations of 1 mg/ml, using solution concentrations of 0, 0.01, 0.1 and 1 mg/ml hyaluronan there were no detectable alterations in the rate of seeded hydroxyapatite growth and proliferation. These data indicate that although in vitro hyaluronan may bind with weak affinity to hydroxyapatite, this interaction does not affect mineral growth, and the presence of hyaluronan would not contribute to the increased inhibitory potential of cartilage proteoglycan aggregate relative to monomer preparations.