Visualization of 3D osteon morphology by synchrotron radiation micro‐CT

Abstract

Cortical bone histology has been the subject of scientific inquiry since the advent of the earliest microscopes. Histology -literally the study of tissue -is a field nearly synonymous with 2D thin sections. That said, progressive developments in high-resolution X-ray imaging are enabling 3D visualization to reach ever smaller structures. Micro-computed tomography (micro-CT), employing conventional X-ray sources, has become the gold standard for 3D analysis of trabecular bone and is capable of detecting the structure of vascular (osteonal) porosity in cortical bone. To date, however, direct 3D visualization of secondary osteons has eluded micro-CT based upon absorption-derived contrast. Synchrotron radiation micro-CT, through greater image quality, resolution and alternative contrast mechanisms (e.g. phase contrast), holds great potential for non-destructive 3D visualization of secondary osteons. Our objective was to demonstrate this potential and to discuss areas of bone research that can be advanced through the application of this approach. We imaged human mid-femoral cortical bone specimens derived from a 20-year-old male (Melbourne Femur Collection) at the Advanced Photon Source synchrotron (Chicago, IL, USA) using the 2BM beam line. A 60-mm distance between the target and the detector was employed to enhance visualization of internal structures through propagation phase contrast. Scan times were 1 h and images were acquired with 1.4-lm nominal isotropic resolution. Computer-aided manual segmentation and volumetric 3D rendering were employed to visualize secondary osteons and porous structures, respectively. Osteonal borders were evident via two contrast mechanisms. First, relatively new (hypomineralized) osteons were evident due to differences in X-ray attenuation relative to the surrounding bone. Second, osteon boundaries (cement lines) were delineated by phase contrast. Phase contrast also enabled the detection of soft tissue remnants within the vascular pores. The ability to discern osteon boundaries in conjunction with vascular and cellular porosity revealed a number of secondary osteon morphologies and provided a unique 3D perspective of the superimposition of secondary osteons on existing structures. Improvements in resolution and optimization of the propagation phase contrast promise to provide further improvements in structural detail in the future.

Does 3D orientation account for variation in osteon morphology assessed by 2D histology?

“…phase contrast) in order to directly evaluate osteon geometry in 3D (Cooper et al. ). A greater appreciation of the morphological variation of osteons opens the door to more rigorous exploration of what factors underpin this variation.…”

“…) or synchrotron‐based micro‐CT imaging (Cooper et al. ). As noted for serial section‐based studies, imaging‐based studies have also revealed great complexity and variation in 3D osteon structure – but they, too, have yielded largely qualitative insights so far.…”

Micro- and Nano-CT for the Study of Bone Ultrastructure

“…During the past decade, SR micro-CT has been used for the simultaneous assessment of structure and mineralization in human or animal trabecular bone [42,43]. Its application to cortical bone also permits quantification of the Haversian canal network and visualize osteons [34,44,45].…”

Role of cortical bone in hip fracture

“…104 New imaging techniques now make it possible to investigate the detailed 3D structure of individual osteons in the femoral cortex and the anatomical relationships between them. 105,106 These offer new avenues for understanding how conversion of the cortex to trabecular bone, leading to cortical thinning, 104,107 develops and to how osteonal structures might help preserve the toughness of bone once a crack threatens to expand beyond the osteon's own territory. 108 Another cause of increased remodelling is the secondary hyperparathyroidism associated with reduced vitamin D levels, commonly seen in the elderly.…”