Multiscale characterization of pathological bone tissue

• Published in: Microscopy research and technique Vol. 85; no. 2; pp. 469 - 486
• Main Authors: Eren, E. Deniz, Nijhuis, Wouter H., Weel, Freek, Dede Eren, Aysegul, Ansari, Sana, Bomans, Paul H.H., Friedrich, Heiner, Sakkers, Ralph J., Weinans, Harrie, With, Gijsbertus
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01-02-2022; Wiley Subscription Services, Inc
• Subjects: Biomedical materials; Bone and Bones; Bone composition; Bone imaging; Bones; Cell interactions; Collagen; Diameters; electron microscopy; electron tomography; Electrons; Fibrils; focused ion beam; Imaging techniques; Imaging, Three-Dimensional; Microscopic analysis; Microscopy, Electron, Transmission; Minerals; optical microscopy; Osteocytes; pathological bone tissues; Sample preparation; serial slice and view; Tissues; Transmission electron microscopy; ultrastructure of bone; focused ion beam; optical microscopy; pathological bone tissues; collagen; electron microscopy; ultrastructure of bone; electron tomography; serial slice and view
• ISSN: 1059-910X; 1097-0029
• DOI: 10.1002/jemt.23920

ABSTRACT
Bone is a complex natural material with a complex hierarchical multiscale organization, crucial to perform its functions. Ultrastructural analysis of bone is crucial for our understanding of cell to cell communication, the healthy or pathological composition of bone tissue, and its three‐dimensional (3D) organization. A variety of techniques has been used to analyze bone tissue. This article describes a combined approach of optical, scanning electron, and transmission electron microscopy for the ultrastructural analysis of bone from the nanoscale to the macroscale, as illustrated by two pathological bone tissues. By following a top‐down approach to investigate the multiscale organization of pathological bones, quantitative estimates were made in terms of calcium content, nearest neighbor distances of osteocytes, canaliculi diameter, ordering, and D‐spacing of the collagen fibrils, and the orientation of intrafibrillar minerals which enable us to observe the fine structural details. We identify and discuss a series of two‐dimensional (2D) and 3D imaging techniques that can be used to characterize bone tissue. By doing so we demonstrate that, while 2D imaging techniques provide comparable information from pathological bone tissues, significantly different structural details are observed upon analyzing the pathological bone tissues in 3D. Finally, particular attention is paid to sample preparation for and quantitative processing of data from electron microscopic analysis.
Multiscale characterization of pathological bone tissues.