The ultrastructure of anorganic bovine bone and selected synthetic hyroxyapatites used as bone graft substitute materials

Abstract

The objective of this study was to investigate the morphology and organization of apatite crystallites in mature mammalian bone. Anorganic bovine bone was studied in this investigation to allow for the examination of the mineral crystallites after removal of the organic phase. Field-emission low-voltage scanning electron microscopy (FE-LVSEM) was employed to obtain images at nanometer resolution without the application of a conductive coating. Transmission electron microscopy (TEM) of the samples was also performed to confirm the identification of features observed in the SEM and to allow for comparison with earlier studies of bone mineral architecture. For comparison, in order to demonstrate how the interaction of collagen and apatite results in the architecture and crystal structure of bone mineral, two synthetic hydroxyapatite materials were also analyzed: OsteoGen and OsteoGraf/LD300. FE-LVSEM revealed distinctive features of bone mineral: a fibrillar organization of crystallites, a periodic spacing of crystallites along the fibrils consistent with the banding pattern of collagen, inter-fibrillar bridging crystallites, and a plate-like habit of the crystallites. These findings supported the hypothesis, derived from the earlier TEM data of others, that the mineralization of collagen comprising osteoid proceeds by the formation of apatite crystallites within the fibers at selected periodic sites along their length. Moreover, the very presence in this anorganic material of distinct fibers comprised of the crystallites is demonstration of inter-crystallite bonding. The crystallites of the synthetic hydroxyapatite materials did not display any of these ultrastructural features.