Microcracks in compact bone: a three-dimensional view.

Microcracks have been implicated in the loss of bone quality for osteoporosis. In order to detect and monitor their growth, and to use these data to predict fractures, it is essential to obtain quantitative data regarding their shape in three dimensions. Beam-shaped bone samples from sheep radii were prepared and stained with fluorochrome dyes and tested in cyclical fatigue under four-point bending in a servo-hydraulic fatigue-testing machine. Samples were tested at a frequency of 30 Hz under load control at a stress range of 100 MPa. Holes were drilled into bone samples and used as reference points for reconstructions. A series of thin longitudinal sections were cut using a sledge macrotome. A two-dimensional image of each section was examined using an epifluorescence microscope and images transferred to a PC via a CCD low-light colour video camera. A three-dimensional image of each microcrack was reconstructed using computer software, and its dimensions measured. Cracks were elliptical in shape, longer in the longitudinal direction and with a mean aspect ratio of 5.5 +/- 1.05. The mean (+/- SD) length and width of labelled microcracks were 488 +/- 151 and 88 +/- 21 microm, respectively.