Collagen fibre orientation in bovine secondary osteons by collagenase etching

The orientation of collagen fibres in bovine secondary osteons has been investigated in the scanning electron microscope (SEM) by removal independently of firstly the mineral component and secondly the collagen fibres. Demineralization of polished transverse sections reveals a lamellar structure for the collagen component but the precise orientation of the collagen in each ring is not unequivocably determined. However, by using a collagenase solution to etch away the collagen component of a polished surface, holes are produced in the mineral revealing the former position of the fibres. The greater rigidity of the mineral component ensures that the structure does not collapse and produce artifacts. A specimen cut so that transverse and longitudinal sections are simultaneously observed allows the relationship between the structural features on each surface to be revealed. Analysis of such micrographs indicates a model for the collagen component of osteons in which the lamellar structure contains fibres with orientations alternately parallel to and circumferential to the long axis of the osteon. Tilting the samples to look directly down the holes shows that the fibres are not precisely longitudinal and circumferential but are tilted from these ideals by a variable angle (typically 20 degrees) the precise angle probably being an important factor related to the in vivo mechanical property requirements.