Bone strength and failure are increasingly thought to be due to ultrastructural properties, such as the morphology of the lacuno-canalicular network, the collagen fiber orientation and the mineralization on the nanoscale. However, these properties have not been studied in 3D so far. Here we report the investigation of the human bone ultrastructure with X-ray phase nanotomography, which now provides the required sensitivity, spatial resolution and field of view. The 3D organization of the lacuno-canalicular network is studied in detail over several cells in osteonal and interstitial tissue. Nanoscale density variations are revealed and show that the cement line separating these tissues is hypermineralized. Finally, we show that the collagen fibers are organized as a twisted plywood structure in 3D.

In bone remodeling, maturation of the newly formed osteonal tissue is associated with a rapid primary increase followed by a slower secondary increase of mineralization. This requires supply and precipitation of mineral into the bone matrix. Mineral delivery can occur only from the extracellular fluid via interfaces such as the Haversian system and the osteocyte pore network. We hypothesized that in mineralization, mineral exchange is achieved by the diffusion of mineral from the lacunar-canalicular network (LCN) to the bone matrix, resulting in a gradual change in tissue mineralization with respect to the distance from the pore-matrix interface. We expected to observe alterations in the mass density distribution with tissue age. We further hypothesized that mineral exchange occurs not only at the lacunar but also at the canalicular boundaries. The aim of this study was, therefore, to investigate the spatial distribution of mass density in the perilacunar and pericanalicular bone matrix and to explore how these densities are influenced by tissue aging. This is achieved by analyzing human jawbone specimens originating from four healthy donors and four treated with high-dosage bisphosphonate using synchrotron radiation phase-contrast nano-CT with a 50-nm voxel size. Our results provide the first experimental evidence that mass density in the direct vicinity of both lacunae (p < 0.001) and canaliculi (p < 0.001) is different from the mean matrix mass density, resulting in gradients with respect to the distance from both pore-matrix interfaces, which diminish with increasing tissue age. Though limited by the sample size, these findings support our hypotheses. Moreover, the density gradients are more pronounced around the lacunae than around the canaliculi, which are explained by geometrical considerations in the LCN morphology. In addition, we speculate that mineral exchange occurs at all interfaces of the LCN, not only in mineralization but also in mineral homeostasis. © 2014 American Society for Bone and Mineral Research.

In computed tomography (CT), projection images of the sample are acquired over an angular range between 180 to 360 degrees around a rotation axis. A special case of CT is that of limited-angle CT, where some of the rotation angles are inaccessible, leading to artefacts in the reconstrucion because of missing information. The case of flat samples is considered, where the projection angles that are close to the sample surface are either i) completely unavailable or ii) very noisy due to the limited transmission at these angles. Computed laminography (CL) is an imaging technique especially suited for flat samples. CL is a generalization of CT that uses a rotation axis tilted by less than 90 degrees with respect to the incident beam. Thus CL avoids using projections from angles closest to the sample surface. We make a quantitative comparison of the imaging artefacts between CL and limited-angle CT for the case of a parallel-beam geometry. Both experimental and simulated images are used to characterize the effect of the artefacts on the resolution and visible image features. The results indicate that CL has an advantage over CT in cases when the missing angular range is a significant portion of the total angular range. In the case when the quality of the projections is limited by noise, CT allows a better tradeoff between the noise level and the missing angular range.