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Atomic force microscopy and indentation force measurement of bone

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Abstract This review is summarizing the results obtained from atomic force microscopy (AFM) and nanoindentation experiments to date. The combination of both techniques is especially powerful. It allows to carefully choose indentation locations as well as the post‐hoc analysis of the created indents, and hence the possibility to assess the properties of microstructural elements of bonessue. In addition, AFM has improved our understanding of bone ultrastructure and force spectroscopy experiments have led to the discovery of a molecular self‐healing effect of bone that may be based on a small fraction of unstructured proteins. Nanoindentation measurements on bone, pose inherent problems since bone is an anisotropic solid showing elastic, viscoelastic, and time‐dependent plastic behavior. Hence, derived parameters such as elastic modulus and hardness are to some extent dependent on measurement protocols. However, the development of extensions to the Oliver–Pharr method, being the most widely used analysis method, as well as novel dynamic testing techniques could improve the situation. Nanoindentation is widely used to study bone tissue and some important principal findings have been reported to date. These are presented here together with specific results from nanoindentation experiments of human and animal bones and tables are presented collating the data that can be found in the literature to date. Copyright © 2009 John Wiley & Sons, Inc. This article is categorized under: Nanotechnology Approaches to Biology > Nanoscale Systems in Biology

Atomic force microscopy tapping mode images of (a) collagen type I fibrils on a native bone surface,37 (b) hydroxyapatite mineral crystals extracted from bone,38 and (c) a dried film of purified osteopontin on a mica surface.39.

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Residual indentation in a bovine cortical bone90 (a) and a collagen fibril from a rat‐tail tendon104 (b).

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Atomic force microscopy images of a bone lamella without64 (a) and with101 (b) indentations. The change in elastic modulus of individual lamellae when from the center to the periphery of an osteon is shown in (c), data from Ref. 63.

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Typical nanoindentation data from an experiment with52 (a) and without61 (b) hold time at maximum load, as well as a schematic representation explaining the analysis of the unloading part of the curve using the Oliver–Pharr method105 (c). The variation of the anisotropic indentation modulus compared to the true modulus of bone, both calculated from ultrasonic measurement data,108 is shown in (d).

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Fracture surface of trabecular bovine bone exhibiting collagen fibrils coated with hydroxyapatite particles (a) and the same surface location after treatment with ethylenediamine tetraacetic acid (EDTA) (b), exposing the typical D‐banding pattern of collagen type I fibrils.51 Unstructured organic matrix (white arrows) as seen in a similar atomic force microscopy image of trabecular bovine bone87 (c), and a nanoscale model of bone (d).

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