The interfacial strength of secondary osteons from the diaphysis of the Thoroughbred equine third metacarpal was evaluated using the fiber pushout test. The pushout was performed on 300-500 microm sections of 4x4x15 mm bone blocks machined from four anatomic regions of the cortex. Pushout strength was evaluated from proximal to distal location within the diaphysis on four osteon types classified under polarized light on adjacent histologic sections from each block. The shear strength of the interfaces were estimated from shear lag theory. Differences were found in the interfacial strength of osteons based on appearance under polarized light with bright field having the highest interfacial strength (40.3 MPa). The lowest strength was found in the dark field osteons (22.8 MPa). The dorsal region had the highest shear strength and toughness compared to all other regions. The cement line and interlamellar interfaces are similar in strength, but exhibit regional dependence--specifically, the palmar region strength is less (17.5 MPa) than the osteon interlamellar interfaces (30.4 MPa) and osteon type dependent (alternating significantly weaker than other types). Histomorphometry revealed significant regional differences (p<0.0001) in osteon area fraction among the four osteon types as well as differences in the osteon diameter (p=0.01), with dorsal regions having larger osteons (170 microm) than the palmar region (151 microm). Fatigue life and fracture toughness of Haversian bone are reported in the literature to be regionally dependent and are known to be associated with osteon pullout--an osteon interfacial phenomenon. Therefore, the results presented in this study are important to further the understanding of the mechanisms of fragility and damage accumulation in cortical bone.

Stress fractures are overuse injuries of bone that affect elite athletes and military recruits. One response of cortical bone to stress fracture is to lay down periosteal callus. The objectives of this study were to determine if material properties are different among bones with different stages of stress fracture callus, at both a callus site and at a distal site. Cortical specimens were mechanically tested to determine their stress-strain response. Material property differences were examined using nonparametric and regression analyses. At the callus site, material properties were low during the earliest stages of callus, higher with increasing callus maturity, but dropped at the late stage of callus. At the distal site, the material properties were low during early stages of callus and approached, or returned to, those of bones without callus during the late stages of callus. The effects of stress fracture and bone callus are not limited to the focal site of stress fracture.

SummaryWe describe a novel automated technique for visualizing the three-dimensional distribution of fluorochrome-labelled components, in which image resolution is uncoupled from specimen size. This method is based on computer numerically controlled milling technology and combines an arrayed imaging technique with fluorescence capabilities. Fluorescent signals are segmented by emission spectra such that multiple fluorochromes present within a single specimen may be reconstructed and visualized individually or as a group. The automated nature of the system minimizes the workload and time involved in image capture and volume reconstruction. As an application, the system was used to image zones of fluorochrome-labelled microdamage within an 8-mm diameter cylinder of trabecular bone at a voxel size of 3 × 3 × 8 µm 3 . Our reconstruction of this specimen provides a visual map and quantitative measures of the volume of damage present throughout the cylinder, clearly demonstrating the interpretive power afforded by threedimensional visualization. The three-dimensional nature of this highly automated and adaptable system has the potential to facilitate new diagnostic tools and techniques with application to a wide range of biological and medical research fields.

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