Hydroxyapatite formation in the presence of proteoglycans of reduced sulfate content: studies in the brachymorphic mouse

Abstract

Proteoglycans from the brachymorphic (bm/bm) mouse have a reduced sulfate content due to the impaired activity of adenosine phosphosulfate phosphokinase in these animals. X-ray diffraction and infrared analyses of the mineral from the calcified cartilage of the bm/bm mice demonstrate the presence of significantly larger and more perfect hydroxyapatite crystals of lower carbonate to phosphate content than crystals found in the control animals. No differences were seen in the mineral content, crystallite size, CO3:PO4 ratio, or infrared splitting factors measured in the diaphyseal bone from these animals. Electron microscopic examination similarly shows larger, more disorganized crystals in the bm/bm animals' calcified cartilage as contrasted with controls. In vitro, proteoglycan aggregates from these dwarf mice are shown in a collagen gel-growth system to be less effective inhibitors of hydroxyapatite formation and growth than similarly size sulfated proteoglycans from age-matched control animals. The proteoglycans from the control mice were comparable in inhibitory ability to proteoglycan aggregates extracted from fetal bovine epiphyses. The in vitro and in vivo mineral parameters suggest the importance of sulfate for the interaction between proteoglycans and mineral in growth plate calcification.