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50 years of scanning electron microscopy of bone—a comprehensive overview of the important discoveries made and insights gained into bone material properties in health, disease, and taphonomy

Shah, Furqan A., Ruscsák, Krisztina, Palmquist, Anders·2019·Bone Research
DOI 10.1038/s41413-019-0053-zPMC PMC6531483
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Abstract

Bone is an architecturally complex system that constantly undergoes structural and functional optimisation through renewal and repair. The scanning electron microscope (SEM) is among the most frequently used instruments for examining bone. It offers the key advantage of very high spatial resolution coupled with a large depth of field and wide field of view. Interactions between incident electrons and atoms on the sample surface generate backscattered electrons, secondary electrons, and various other signals including X-rays that relay compositional and topographical information. Through selective removal or preservation of specific tissue components (organic, inorganic, cellular, vascular), their individual contribution(s) to the overall functional competence can be elucidated. With few restrictions on sample geometry and a variety of applicable sample-processing routes, a given sample may be conveniently adapted for multiple analytical methods. While a conventional SEM operates at high vacuum conditions that demand clean, dry, and electrically conductive samples, non-conductive materials (e.g., bone) can be imaged without significant modification from the natural state using an environmental scanning electron microscope. This review highlights important insights gained into bone microstructure and pathophysiology, bone response to implanted biomaterials, elemental analysis, SEM in paleoarchaeology, 3D imaging using focused ion beam techniques, correlative microscopy and in situ experiments. The capacity to image seamlessly across multiple length scales within the meso-micro-nano-continuum, the SEM lends itself to many unique and diverse applications, which attest to the versatility and user-friendly nature of this instrument for studying bone. Significant technological developments are anticipated for analysing bone using the SEM.

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Figures

Fig. 1 Parts of a scanning electron microscope (SEM) and the typical signals that are recorded from bone. BSE backscattered electrons, SE secondary electrons, EDX energy-dispersive X-ray spectroscopy
Fig. 1 Parts of a scanning electron microscope (SEM) and the typical signals that are recorded from bone. BSE backscattered electrons, SE secondary electrons, EDX energy-dispersive X-ray spectroscopyCC BY · PubMed Central
Fig. 2 Imaging bone in the SEM. a BSE-SEM photomontage of a human rib viewed in cross-section. Local variations in mineralisation density, Haversian canals, resorption spaces, and osteocyte lacunae can be detected. From Bereshiem et al. Adapted with permission from John Wiley and Sons. Copyright 2019 7. b Osteocyte lacunar density determined using BSE imaging. From Bach-Gansmo et al.12. Adapted with permission from Elsevier. Copyright 201512. c A cement line between osteonal bone and interstitial bone. From Skedros et al. Adapted with permission from John Wiley and Sons. Copyright 200520. d The intertrabecular spaces in antler bone are occupied by primary osteons. Trabeculae (asterisks) and unremodelled islands of calcified cartilage (arrow) can be identified. From Kierdorf et al. Adapted with permission from John Wiley and Sons. Copyright 201330. e Topographical BSE-SEM. For each 90° sector of an annular BSE detector, a separate image is recorded containing information sensitive to the direction of apparent illumination (arrows). From Boyde A. Adapted with permission from John Wiley and Sons. Copyright 200331. f Osteon pull-out under cyclic mechanical loading observed using SE imaging. From Hiller et al. Adapted with permission from John Wiley and Sons. Copyright 200347. g Cell surface detail of osteoblasts on the surface of parietal bone. From Jones S. J. Adapted with permission from Springer Nature. Copyright 197457. h Osteoblasts appear to organise collagen fibrils through flat basal processes. From Pazzaglia U.E. et al. Adapted with permission from Springer Nature. Copyright 201061. i Hypermineralised osteocyte lacuna containing mineralised apoptotic debris. From Shah et al. Adapted with permission from the American Chemical Society. Copyright 2017.69 j Severely disorganised bone microstructure in melorheostosis. From Fratzl-Zelman et al. Adapted with permission from John Wiley and Sons. Copyright 201972. k Bone with (right) and without (left) osteocytes. From Atkins et al. Adapted with permission from the National Academy of Sciences. Copyright 201477. l An osteocyte and associated canalicular network exposed by resin cast etching. From Feng et al. Adapted with permission from John Wiley and Sons. Copyright 200679. m Howship’s lacunae: longitudinally extended resorption (LER; left) and reticulate patch resorption (RPR; right) lacunae. From Gentzsch et al. Adapted with permission from Springer Nature. Copyright 200384. n Cryo-SEM. The combination of SE (above) and BSE (below) imaging provides morphological and compositional information. From Mahamid et al. Adapted with permission from the National Academy of Sciences. Copyright 201097
Fig. 2 Imaging bone in the SEM. a BSE-SEM photomontage of a human rib viewed in cross-section. Local variations in mineralisation density, Haversian canals, resorption spaces, and osteocyte lacunae can be detected. From Bereshiem et al. Adapted with permission from John Wiley and Sons. Copyright 2019 7. b Osteocyte lacunar density determined using BSE imaging. From Bach-Gansmo et al.12. Adapted with permission from Elsevier. Copyright 201512. c A cement line between osteonal bone and interstitial bone. From Skedros et al. Adapted with permission from John Wiley and Sons. Copyright 200520. d The intertrabecular spaces in antler bone are occupied by primary osteons. Trabeculae (asterisks) and unremodelled islands of calcified cartilage (arrow) can be identified. From Kierdorf et al. Adapted with permission from John Wiley and Sons. Copyright 201330. e Topographical BSE-SEM. For each 90° sector of an annular BSE detector, a separate image is recorded containing information sensitive to the direction of apparent illumination (arrows). From Boyde A. Adapted with permission from John Wiley and Sons. Copyright 200331. f Osteon pull-out under cyclic mechanical loading observed using SE imaging. From Hiller et al. Adapted with permission from John Wiley and Sons. Copyright 200347. g Cell surface detail of osteoblasts on the surface of parietal bone. From Jones S. J. Adapted with permission from Springer Nature. Copyright 197457. h Osteoblasts appear to organise collagen fibrils through flat basal processes. From Pazzaglia U.E. et al. Adapted with permission from Springer Nature. Copyright 201061. i Hypermineralised osteocyte lacuna containing mineralised apoptotic debris. From Shah et al. Adapted with permission from the American Chemical Society. Copyright 2017.69 j Severely disorganised bone microstructure in melorheostosis. From Fratzl-Zelman et al. Adapted with permission from John Wiley and Sons. Copyright 201972. k Bone with (right) and without (left) osteocytes. From Atkins et al. Adapted with permission from the National Academy of Sciences. Copyright 201477. l An osteocyte and associated canalicular network exposed by resin cast etching. From Feng et al. Adapted with permission from John Wiley and Sons. Copyright 200679. m Howship’s lacunae: longitudinally extended resorption (LER; left) and reticulate patch resorption (RPR; right) lacunae. From Gentzsch et al. Adapted with permission from Springer Nature. Copyright 200384. n Cryo-SEM. The combination of SE (above) and BSE (below) imaging provides morphological and compositional information. From Mahamid et al. Adapted with permission from the National Academy of Sciences. Copyright 201097CC BY · PubMed Central
Fig. 3 Sources of osteogenesis around implant biomaterials. 1. Contact osteogenesis. 2. Distance osteogenesis. 3. De novo formed woven bone. 4. Autogenous bone fragments
Fig. 3 Sources of osteogenesis around implant biomaterials. 1. Contact osteogenesis. 2. Distance osteogenesis. 3. De novo formed woven bone. 4. Autogenous bone fragmentsCC BY · PubMed Central
Fig. 4 Imaging bone around implant biomaterials. a BSE imaging of bone formed around laser-ablated titanium. From Palmquist et al. Adapted with permission from John Wiley and Sons. Copyright 2011115. b BSE imaging of HAp-coated titanium implants. From Merolli et al. Adapted by permission from Springer Nature. Copyright 2000132. c Local bisphosphonate delivery (BP; 16 µg/implant. Ctrl; 0 µg/implant) from HAp-coated cp-Ti implants (3 mm diameter) promotes bone formation. From Peter et al. Adapted with permission from John Wiley and Sons. Copyright 2006135. d Ingrowth of mineralised tissue into 3D printed polycaprolactone + β-tricalcium phosphate (80:20) scaffolds with a repeating 0°/90° strut laydown pattern. From Paris et al. Adapted with permission from Elsevier. Copyright 2017153. e Resin cast etching for direct visualisation of osteocyte attachment to various implant surfaces. Example #1: Ti6Al4V. From Shah et al. Adapted with permission from Elsevier. Copyright 201674. Example #2: CoCr. From Shah et al. Adapted with permission from John Wiley and Sons. Copyright 201873
Fig. 4 Imaging bone around implant biomaterials. a BSE imaging of bone formed around laser-ablated titanium. From Palmquist et al. Adapted with permission from John Wiley and Sons. Copyright 2011115. b BSE imaging of HAp-coated titanium implants. From Merolli et al. Adapted by permission from Springer Nature. Copyright 2000132. c Local bisphosphonate delivery (BP; 16 µg/implant. Ctrl; 0 µg/implant) from HAp-coated cp-Ti implants (3 mm diameter) promotes bone formation. From Peter et al. Adapted with permission from John Wiley and Sons. Copyright 2006135. d Ingrowth of mineralised tissue into 3D printed polycaprolactone + β-tricalcium phosphate (80:20) scaffolds with a repeating 0°/90° strut laydown pattern. From Paris et al. Adapted with permission from Elsevier. Copyright 2017153. e Resin cast etching for direct visualisation of osteocyte attachment to various implant surfaces. Example #1: Ti6Al4V. From Shah et al. Adapted with permission from Elsevier. Copyright 201674. Example #2: CoCr. From Shah et al. Adapted with permission from John Wiley and Sons. Copyright 201873CC BY · PubMed Central
Fig. 5 Elemental analysis. a BSE image and Ca (magenta), P (yellow), and Sr (cyan) elemental maps demonstrate Sr incorporation into the fracture callus after therapeutic administration. From Brüel et al. Adapted with permission from Springer Nature. Copyright 2011166. b Colour-merge image C (red), Ca (green), and Ti (blue) elemental maps reveals highly mineralised bone around laser-ablated titanium implants. From Palmquist et al. Adapted with permission from John Wiley and Sons. Copyright 2011115
Fig. 5 Elemental analysis. a BSE image and Ca (magenta), P (yellow), and Sr (cyan) elemental maps demonstrate Sr incorporation into the fracture callus after therapeutic administration. From Brüel et al. Adapted with permission from Springer Nature. Copyright 2011166. b Colour-merge image C (red), Ca (green), and Ti (blue) elemental maps reveals highly mineralised bone around laser-ablated titanium implants. From Palmquist et al. Adapted with permission from John Wiley and Sons. Copyright 2011115CC BY · PubMed Central
Fig. 6 Selective removal and/or preservation of specific tissue components. a Osteonal lamellar pattern enhanced by etching with citric acid. From Congiu and Pazzaglia. Adapted with permission from John Wiley and Sons. Copyright 2011.176 b Osteoblastic-osteocyte lacunae on the surface of trabecular bone treated with NaOCl. From Shah et al. Adapted with permission from Springer Nature. Copyright 2016.60 c Heat-deproteinised and fractured surface. Ordered layout of mineral crystal aggregates arranged in a concentric sequence of crests and grooves. From Pazzaglia et al. Adapted with permission from John Wiley and Sons. Copyright 2016.180 d After OsO4 and K4Fe(CN)6 treatment, a portion of lining cells is detached by ultrasonication and the underlying surface is exposed. Two morphological types of cells are recognised here: (i) convex dome-shaped cells with a non-adhering border (denoted as “ostC1”; mean surface area of 52.5 µm2 per cell), and (ii) flattened cells on the bone surface with spreading equatorial, cytoplasmic processes (denoted as “ostC2”; mean surface area of 179 µm2 per cell). From Pazzaglia et al. Adapted with permission from John Wiley and Sons. Copyright 2014.59 e Multi-layered cast of the osteocyte network and vasculature obtained by prolonged, repeated exposure to HCl and KOH solutions. From Pazzaglia and Congiu. Adapted with permission from John Wiley and Sons. Copyright 2013.184 f Directly opposing an implant surface, collagen is exposed by etching with HCl after mechanically separating the implant from a resin embedded bone-implant specimen. From Traini et al. Adapted with permission from John Wiley and Sons. Copyright 2005.103 g Resin cast etching reveals osteocyte attachment to the surface of a laser-ablated cp-Ti implant. Fine topographical features of the implant surface remain intact after H3PO4 and NaOCl exposure. From Shah et al. Adapted with permission from the American Chemical Society. Copyright 2015.159
Fig. 6 Selective removal and/or preservation of specific tissue components. a Osteonal lamellar pattern enhanced by etching with citric acid. From Congiu and Pazzaglia. Adapted with permission from John Wiley and Sons. Copyright 2011.176 b Osteoblastic-osteocyte lacunae on the surface of trabecular bone treated with NaOCl. From Shah et al. Adapted with permission from Springer Nature. Copyright 2016.60 c Heat-deproteinised and fractured surface. Ordered layout of mineral crystal aggregates arranged in a concentric sequence of crests and grooves. From Pazzaglia et al. Adapted with permission from John Wiley and Sons. Copyright 2016.180 d After OsO4 and K4Fe(CN)6 treatment, a portion of lining cells is detached by ultrasonication and the underlying surface is exposed. Two morphological types of cells are recognised here: (i) convex dome-shaped cells with a non-adhering border (denoted as “ostC1”; mean surface area of 52.5 µm2 per cell), and (ii) flattened cells on the bone surface with spreading equatorial, cytoplasmic processes (denoted as “ostC2”; mean surface area of 179 µm2 per cell). From Pazzaglia et al. Adapted with permission from John Wiley and Sons. Copyright 2014.59 e Multi-layered cast of the osteocyte network and vasculature obtained by prolonged, repeated exposure to HCl and KOH solutions. From Pazzaglia and Congiu. Adapted with permission from John Wiley and Sons. Copyright 2013.184 f Directly opposing an implant surface, collagen is exposed by etching with HCl after mechanically separating the implant from a resin embedded bone-implant specimen. From Traini et al. Adapted with permission from John Wiley and Sons. Copyright 2005.103 g Resin cast etching reveals osteocyte attachment to the surface of a laser-ablated cp-Ti implant. Fine topographical features of the implant surface remain intact after H3PO4 and NaOCl exposure. From Shah et al. Adapted with permission from the American Chemical Society. Copyright 2015.159CC BY · PubMed Central
Fig. 7 SEM in paleoarchaeology. a Defects of active osteomalacia visible in archaeological bone from the adult rib. Multiple areas of incomplete mineralisation (IM) and defect cement lines (DCL) are noted. From Brickley et al. Adapted with permission from John Wiley and Sons. Copyright 2007.195 b Pyrite deposits within Haversian canals in human tibia from 2 000 years ago. From Tjelldén et al. Adapted with permission from John Wiley and Sons. Copyright 2018.202 c A number of high-density foci within a single secondary osteonal system in archaeological human tibia (left). Bacterial ingress seen extending from a single osteocyte lacuna. Other osteocytes exhibit demineralisation boundaries or enlargement (right). From Bell LS. 2012. Forensic Microscopy for Skeletal Tissues. Adapted with permission from Springer Nature. Copyright 2012.196 d Mycelia mineralised with Fe/Mn oxides and calcite. Here, the mycelia (white) are seen as sunflower-like aggregates and networks of hyphae filling the resorption canals inside the compact bone tissue. From Owocki et al. Reproduced under the terms of the Creative Commons Attribution License (CC BY 4.0)204
Fig. 7 SEM in paleoarchaeology. a Defects of active osteomalacia visible in archaeological bone from the adult rib. Multiple areas of incomplete mineralisation (IM) and defect cement lines (DCL) are noted. From Brickley et al. Adapted with permission from John Wiley and Sons. Copyright 2007.195 b Pyrite deposits within Haversian canals in human tibia from 2 000 years ago. From Tjelldén et al. Adapted with permission from John Wiley and Sons. Copyright 2018.202 c A number of high-density foci within a single secondary osteonal system in archaeological human tibia (left). Bacterial ingress seen extending from a single osteocyte lacuna. Other osteocytes exhibit demineralisation boundaries or enlargement (right). From Bell LS. 2012. Forensic Microscopy for Skeletal Tissues. Adapted with permission from Springer Nature. Copyright 2012.196 d Mycelia mineralised with Fe/Mn oxides and calcite. Here, the mycelia (white) are seen as sunflower-like aggregates and networks of hyphae filling the resorption canals inside the compact bone tissue. From Owocki et al. Reproduced under the terms of the Creative Commons Attribution License (CC BY 4.0)204CC BY · PubMed Central
Fig. 8 Focused ion beam (FIB) techniques for 3D imaging and sample preparation. a FIB tomography of collagen fibrils in bone. From Reznikov et al. Adapted with permission from the American Association for the Advancement of Science. Copyright 2018.211 b FIB tomography of implanted demineralised dentin matrix and surrounding new bone where osteocytes form an interconnected network of cellular processes. From Tanoue et al. Reproduced under the terms of the Creative Commons Attribution License (CC BY 4.0).216 c Sample preparation for transmission electron microscopy using the in situ lift-out technique, starting with deposition of a 30 µm long, 2 µm wide, and 1 µm-thick protective layer, followed by sequential milling, lift-out, and thinning to electron transparency. From Grandfield et al. Adapted with permission from John Wiley and Sons. Copyright 2012.217 d Sample preparation for ptychographic X-ray computed tomography. From Dierolf et al. Adapted with permission from Springer Nature. Copyright 2010.229 e Micropillars and nanopillars prepared for uniaxial compression testing. From Tertuliano and Greer. Adapted with permission from Springer Nature. Copyright 2016231
Fig. 8 Focused ion beam (FIB) techniques for 3D imaging and sample preparation. a FIB tomography of collagen fibrils in bone. From Reznikov et al. Adapted with permission from the American Association for the Advancement of Science. Copyright 2018.211 b FIB tomography of implanted demineralised dentin matrix and surrounding new bone where osteocytes form an interconnected network of cellular processes. From Tanoue et al. Reproduced under the terms of the Creative Commons Attribution License (CC BY 4.0).216 c Sample preparation for transmission electron microscopy using the in situ lift-out technique, starting with deposition of a 30 µm long, 2 µm wide, and 1 µm-thick protective layer, followed by sequential milling, lift-out, and thinning to electron transparency. From Grandfield et al. Adapted with permission from John Wiley and Sons. Copyright 2012.217 d Sample preparation for ptychographic X-ray computed tomography. From Dierolf et al. Adapted with permission from Springer Nature. Copyright 2010.229 e Micropillars and nanopillars prepared for uniaxial compression testing. From Tertuliano and Greer. Adapted with permission from Springer Nature. Copyright 2016231CC BY · PubMed Central

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Research works printed but not linked: 258

  1. Boyde, A. & Jones, S. J. Scanning electron microscopy of cementum and sharpey fibre bone. Z. Zellforsch. Mikros. Anat. 92, 536–548 (1968). · DOI
  2. Boyde, A. & Hobdell, M. H. Scanning electron microscopy of lamellar bone. Z. Zellforsch. Mikros. Anat. 93, 213–231 (1968). · DOI
  3. Georgiadis, M., Müller, R. & Schneider, P. Techniques to assess bone ultrastructure organization: orientation and arrangement of mineralized collagen fibrils. J. R. Soc. Interface 13, 20160088 (2016). pii. · DOI
  4. Palmquist, A. A multiscale analytical approach to evaluate osseointegration. J. Mater. Sci. Mater. Med. 29, 60 (2018). · DOI
  5. Shah, F. A., Thomsen, P. & Palmquist, A. Osseointegration and current interpretations of the bone-implant interface. Acta Biomater. 84, 1–15 (2019). · DOI
  6. Goldstein, J. I. et al., Scanning Electron Microscopy and X-ray Microanalysis, 1–550 (Springer-Verlag, New York, 2018).
  7. Bereshiem, A. C., Pfeiffer, S. K., Grynpas, M. D. & Alblas, A. Use of backscattered scanning electron microscopy to quantify the bone tissues of midthoracic human ribs. Am. J. Phys. Anthrpol. 168, 262–278 (2019). · DOI
  8. Granke, M., Makowski, A. J., Uppuganti, S. & Nyman, J. S. Prevalent role of porosity and osteonal area over mineralization heterogeneity in the fracture toughness of human cortical bone. J. Biomech. 49, 2748–2755 (2016). · DOI
  9. Zebaze, R. M. et al. Intracortical remodelling and porosity in the distal radius and post-mortem femurs of women: a cross-sectional study. Lancet 375, 1729–1736 (2010). · DOI
  10. Shah, F. A., Stoica, A., Cardemil, C. & Palmquist, A. Multiscale characterization of cortical bone composition, microstructure, and nanomechanical properties in experimentally induced osteoporosis. J. Biomed. Mater. Res A 106(4), 997–1007 (2018). · DOI
  11. Misof, B. M. et al. Intravenous treatment with ibandronate normalizes bone matrix mineralization and reduces cortical porosity after two years in male osteoporosis: a paired biopsy study. J. Bone Miner. Res. 29, 440–449 (2014). · DOI
  12. Bach-Gansmo, F. L. et al. Osteocyte lacunar properties in rat cortical bone: differences between lamellar and central bone. J. Struct. Biol. 191, 59–67 (2015). · DOI
  13. Reid, S. A. & Boyde, A. Changes in the mineral density distribution in human bone with age: Image analysis using backscattered electrons in the SEM. J. Bone Miner. Res. 2, 13–22 (1987). · DOI
  14. Bloebaum, R. D., Skedros, J. G., Vajda, E. G., Bachus, K. N. & Constantz, B. R. Determining mineral content variations in bone using backscattered electron imaging. Bone 20, 485–490 (1997). · DOI
  15. Hoerth, R. M. et al. Correlations between nanostructure and micromechanical properties of healing bone. J. Mech. Behav. Biomed. Mater. 77, 258–266 (2018). · DOI
  16. Cui, F. Z., Zhang, Y., Wen, H. B. & Zhu, X. D. Microstructural evolution in external callus of human long bone. Mater. Sci. Eng. C 11, 27–33 (2000). · DOI
  17. Banse, X. et al. Irreversible perforations in vertebral trabeculae? J. Bone Miner. Res. 18, 1247–1253 (2003). · DOI
  18. Banse, X., Devogelaer, J. P., Holmyard, D. & Grynpas, M. Vertebral cancellous bone turn-over: Microcallus and bridges in backscatter electron microscopy. Micron 36, 710–714 (2005). · DOI
  19. Blaskovic, M. et al. Bone healing following different types of osteotomy: scanning electron microscopy (SEM) and three-dimensional SEM analyses. Microsc. Microanal. 22, 1170–1178 (2016). · DOI
  20. Skedros, J. G., Holmes, J. L., Vajda, E. G. & Bloebaum, R. D. Cement lines of secondary osteons in human bone are not mineral-deficient: new data in a historical perspective. Anat. Rec. A 286A, 781–803 (2005). · DOI
  21. Milovanovic, P. et al. Bone tissue aging affects mineralization of cement lines. Bone 110, 187–193 (2018). · DOI
  22. Bach-Gansmo, F. L., Irvine, S. C., Bruel, A., Thomsen, J. S. & Birkedal, H. Calcified cartilage islands in rat cortical bone. Calcif. Tissue Int. 92, 330–338 (2013). · DOI
  23. Shipov, A. et al. Unremodeled endochondral bone is a major architectural component of the cortical bone of the rat (Rattus norvegicus). J. Struct. Biol. 183, 132–140 (2013). · DOI
  24. Kingsmill, V. J. & Boyde, A. Mineralisation density of human mandibular bone: quantitative backscattered electron image analysis. J. Anat. 192, 245–256 (1998). · DOI
  25. Trivellato, A. E. et al. Osteopetrosis complicated by osteomyelitis of the maxilla and mandible: light and electron microscopic findings. Head. Neck Pathol. 3, 320–326 (2009). · DOI
  26. Boyde, A. et al. A distinctive patchy osteomalacia characterises Phospho1-deficient mice. J. Anat. 231, 298–308 (2017). · DOI
  27. Shabestari, M. et al. Presence of pyrophosphate in bone from an atypical femoral fracture site: A case report. Bone Rep. 6, 81–86 (2017). · DOI
  28. Regelsberger, J. et al. Changes to the cell, tissue and architecture levels in cranial suture synostosis reveal a problem of timing in bone development. Eur. Cell Mater. 24, 441–458 (2012). · DOI
  29. Launey, M. E., Chen, P. Y., McKittrick, J. & Ritchie, R. O. Mechanistic aspects of the fracture toughness of elk antler bone. Acta Biomater. 6, 1505–1514 (2010). · DOI
  30. Kierdorf, U., Flohr, S., Gomez, S., Landete-Castillejos, T. & Kierdorf, H. The structure of pedicle and hard antler bone in the European roe deer (Capreolus capreolus): a light microscope and backscattered electron imaging study. J. Anat. 223, 364–384 (2013). · DOI
  31. Boyde, A. Improved digital SEM of cancellous bone: scanning direction of detection, through focus for in-focus and sample orientation. J. Anat. 202, 183–194 (2003). · DOI
  32. Marks, S. C. Jr, Cielinski, M. J. & Sundquist, K. T. Bone surface morphology reflects local skeletal metabolism. Microsc Res Tech. 33, 121–127 (1996). · DOI
  33. Jones, S. J., Boyde, A. & Pawley, J. B. Osteoblasts and collagen orientation. Cell Tissue Res 159, 73–80 (1975). · DOI
  34. Reid, S. A. Micromorphological characterisation of normal human bone surfaces as a function of age. Scanning Microsc 1, 579–597 (1987).
  35. Mosekilde, L. Consequences of the remodelling process for vertebral trabecular bone structure: a scanning electron microscopy study (uncoupling of unloaded structures). Bone Miner. 10, 13–35 (1990). · DOI
  36. Jayasinghe, J. A. P., Jones, S. J. & Boyde, A. Scanning electron microscopy of human lumbar vertebral trabecular bone surfaces. Virchows Arch. A 422, 25–34 (1993). · DOI
  37. Pazzaglia, U. E., Congiu, T., Raspanti, M., Ranchetti, F. & Quacci, D. Anatomy of the Intracortical Canal system: scanning electron microscopy study in Rabbit Femur. Clin. Orthop. Relat. Res 467, 2446–2456 (2009). · DOI
  38. Corondan, G. & Haworth, W. L. A fractographic study of human long bone. J. Biomech. 19, 207–218 (1986). · DOI
  39. Tomlin, J. L., Lawes, T. J., Blunn, G. W., Goodship, A. E. & Muir, P. Fractographic examination of racing greyhound central (navicular) tarsal bone failure surfaces using scanning electron microscopy. Calcif. Tissue Int 67, 260–266 (2000). · DOI
  40. Weiner, S., Arad, T. & Traub, W. Crystal organization in rat bone lamellae. FEBS Lett. 285, 49–54 (1991). · DOI
  41. Braidotti, P., Bemporad, E., D'Alessio, T., Sciuto, S. A. & Stagni, L. Tensile experiments and SEM fractography on bovine subchondral bone. J. Biomech. 33, 1153–1157 (2000). · DOI
  42. Zhang, W., Tekalur, S. A., Baumann, M. & McCabe, L. R. The effects of damage accumulation on the tensile strength and toughness of compact bovine bone. J. Biomech. 46, 964–972 (2013). · DOI
  43. Wise, L. M., Wang, Z. & Grynpas, M. D. The use of fractography to supplement analysis of bone mechanical properties in different strains of mice. Bone 41, 620–630 (2007). · DOI
  44. Braidotti, P., Branca, F. P. & Stagni, L. Scanning electron microscopy of human cortical bone failure surfaces. J. Biomech. 30, 155–162 (1997). · DOI
  45. Vashishth, D., Tanner, K. E. & Bonfield, W. Contribution, development and morphology of microcracking in cortical bone during crack propagation. J. Biomech. 33, 1169–1174 (2000). · DOI · Paper page
  46. Piekarski, K. Fracture of Bone. J. Appl. Phys. 41, 215–223 (1970). · DOI · Paper page
  47. Hiller, L. P. et al. Osteon pullout in the equine third metacarpal bone: effects of ex vivo fatigue. J. Orthop. Res. 21, 481–488 (2003). · DOI
  48. Li, S., Abdel-Wahab, A. & Silberschmidt, V. V. Analysis of fracture processes in cortical bone tissue. Eng. Fract. Mech. 110, 448–458 (2013). · DOI
  49. Fantner, G. E. et al. Sacrificial bonds and hidden length dissipate energy as mineralized fibrils separate during bone fracture. Nat. Mater. 4, 612–616 (2005). · DOI · Paper page
  50. Nalla, R. K., Kinney, J. H. & Ritchie, R. O. Mechanistic fracture criteria for the failure of human cortical bone. Nat. Mater. 2, 164–168 (2003). · DOI
  51. Tang, T. et al. Shear deformation and fracture of human cortical bone. Bone 71, 25–35 (2015). · DOI
  52. Utku, F. S., Klein, E., Saybasili, H., Yucesoy, C. A. & Weiner, S. Probing the role of water in lamellar bone by dehydration in the environmental scanning electron microscope. J. Struct. Biol. 162, 361–367 (2008). · DOI
  53. Jimenez-Palomar, I., Shipov, A., Shahar, R. & Barber, A. H. Influence of SEM vacuum on bone micromechanics using in situ AFM. J. Mech. Behav. Biomed. Mater. 5, 149–155 (2012). · DOI
  54. Moffat, K. L. et al. Characterization of the structure–function relationship at the ligament-to-bone interface. Proc. Natl Acad. Sci. USA 105, 7947 (2008). · DOI
  55. Cury, D. P., Dias, F. J., Miglino, M. A. & Watanabe, I. S. Structural and ultrastructural characteristics of bone–tendon junction of the calcaneal tendon of adult and elderly wistar rats. PLoS ONE 11, e0153568 (2016). · DOI
  56. Boyde, A. Scanning electron microscopy of bone. Methods Mol. Biol. 816, 365–400 (2012). · DOI
  57. Jones, S. J. Secretory territories and rate of matrix production of osteoblasts. Calcif. Tissue Res. 14, 309–315 (1974). · DOI
  58. Menton, D. N., Simmons, D. J., Chang, S. L. & Orr, B. Y. From bone lining cell to osteocyte—an SEM study. Anat. Rec. 209, 29–39 (1984). · DOI
  59. Pazzaglia, U. E., Congiu, T., Sibilia, V. & Quacci, D. Osteoblast-osteocyte transformation. A SEM densitometric analysis of endosteal apposition in rabbit femur. J. Anat. 224, 132–141 (2014). · DOI
  60. Shah, F. A., Zanghellini, E., Matic, A., Thomsen, P. & Palmquist, A. The orientation of nanoscale apatite platelets in relation to osteoblastic-osteocyte lacunae on trabecular bone surface. Calcif. Tissue Int 98, 193–205 (2016). · DOI
  61. Pazzaglia, U. E., Congiu, T., Marchese, M. & Dell'Orbo, C. The shape modulation of osteoblast-osteocyte transformation and its correlation with the fibrillar organization in secondary osteons: a SEM study employing the graded osmic maceration technique. Cell Tissue Res. 340, 533–540 (2010). · DOI
  62. Qing, H. et al. Demonstration of osteocytic perilacunar/canalicular remodeling in mice during lactation. J. Bone Miner. Res. 27, 1018–1029 (2012). · DOI · Paper page
  63. Kaya, S. et al. Lactation-induced changes in the volume of osteocyte lacunar-canalicular space alter mechanical properties in cortical bone tissue. J. Bone Miner. Res. 32, 688–697 (2017). · DOI
  64. Skedros, J. G. Osteocyte lacuna population densities in sheep, elk and horse calcanei. Cells Tissues Organs 181, 23–37 (2005). · DOI
  65. Busse, B. et al. Decrease in the osteocyte lacunar density accompanied by hypermineralized lacunar occlusion reveals failure and delay of remodeling in aged human bone. Aging cell 9, 1065–1075 (2010). · DOI
  66. Boyde, A. The real response of bone to exercise. J. Anat. 203, 173–189 (2003). · DOI
  67. Jones, S. J., Glorieux, F. H., Travers, R. & Boyde, A. The microscopic structure of bone in normal children and patients with osteogenesis imperfecta: a survey using backscattered electron imaging. Calcif. Tissue Int. 64, 8–17 (1999). · DOI
  68. Carpentier, V. T. et al. Increased proportion of hypermineralized osteocyte lacunae in osteoporotic and osteoarthritic human trabecular bone: implications for bone remodeling. Bone 50, 688–694 (2012). · DOI
  69. Shah, F. A. et al. Micrometer-sized magnesium whitlockite crystals in micropetrosis of bisphosphonate-exposed human alveolar bone. Nano Lett. 17, 6210–6216 (2017). · DOI
  70. Rolvien, T. et al. Early bone tissue aging in human auditory ossicles is accompanied by excessive hypermineralization, osteocyte death and micropetrosis. Sci. Rep. 8, 1920 (2018). · DOI
  71. Bell, L. S., Kayser, M. & Jones, C. The mineralized osteocyte: a living fossil. Am. J. Phys. Anthrpol. 137, 449–456 (2008). · DOI
  72. N. Fratzl-Zelman et al. Melorheostotic bone lesions caused by somatic mutations in MAP2K1 have deteriorated microarchitecture and periosteal reaction. J. Bone Miner. Res. https://doi.org/10.1002/jbmr.3656 (2019). · DOI
  73. Shah, F. A., Jergéus, E., Chiba, A. & Palmquist, A. Osseointegration of 3D printed microalloyed CoCr implants—addition of 0.04% Zr to CoCr does not alter bone material properties. J. Biomed. Mater. Res. A 106, 1655–1663 (2018). · DOI
  74. Shah, F. A., Snis, A., Matic, A., Thomsen, P. & Palmquist, A. 3D printed Ti6Al4V implant surface promotes bone maturation and retains a higher density of less aged osteocytes at the bone-implant interface. Acta Biomater. 30, 357–367 (2016). · DOI
  75. Shah, F. A. et al. Long-term osseointegration of 3D printed CoCr constructs with an interconnected open-pore architecture prepared by electron beam melting. Acta Biomater. 36, 296–309 (2016). · DOI
  76. Shah, F. A., Thomsen, P. & Palmquist, A. A review of the impact of implant biomaterials on osteocytes. J. Dent. Res. 97, 977–986 (2018). · DOI
  77. Atkins, A. et al. Remodeling in bone without osteocytes: billfish challenge bone structure-function paradigms. Proc. Natl Acad. Sci. USA 111, 16047–16052 (2014). · DOI
  78. Kubek, D. J., Gattone, V. H. & Allen, M. R. Methodological assessment of acid-etching for visualizing the osteocyte lacunar-canalicular networks using scanning electron microscopy. Microsc. Res. Tech. 73, 182–186 (2010). · DOI
  79. Rangiani, A. et al. Protective roles of DMP1 in high phosphate homeostasis. PLoS ONE 7, e42329 (2012). · DOI
  80. Zuo, G. L. et al. Activation of HIFa pathway in mature osteoblasts disrupts the integrity of the osteocyte/canalicular network. PLoS ONE 10, e0121266 (2015). · DOI
  81. Zhang, D. et al. Retention of osteocytic micromorphology by sclerostin antibody in a concurrent ovariectomy and functional disuse model. Ann. N. Y. Acad. Sci. 1442, 91–103 (2019). · DOI
  82. Shah, F. A. et al. Laser-modified surface enhances osseointegration and biomechanical anchorage of commercially pure titanium implants for bone-anchored hearing systems. PLoS ONE 11, e0157504 (2016). · DOI
  83. Gentzsch, C., Delling, G. & Kaiser, E. Microstructural classification of resorption lacunae and perforations in human proximal femora. Calcif. Tissue Int. 72, 698–709 (2003). · DOI
  84. Gentzsch, C., Pueschel, K., Deuretzbacher, G., Delling, G. & Kaiser, E. First inventory of resorption lacunae on rods and plates of trabecular bone as observed by scanning electron microscopy. Calcif. Tissue Int. 76, 154–162 (2005). · DOI
  85. Ren, S., Takano, H. & Abe, K. Two types of bone resorption lacunae in the mouse parietal bones as revealed by scanning electron microscopy and histochemistry. Arch. Histol. Cytol. 68, 103–113 (2005). · DOI
  86. Fuller, K., Thong, J. T., Breton, B. C. & Chambers, T. J. Automated three-dimensional characterization of osteoclastic resorption lacunae by stereoscopic scanning electron microscopy. J. Bone Miner. Res. 9, 17–23 (1994). · DOI
  87. Roschger, P., Fratzl, P., Eschberger, J. & Klaushofer, K. Validation of quantitative backscattered electron imaging for the measurement of mineral density distribution in human bone biopsies. Bone 23, 319–326 (1998). · DOI
  88. Roschger, P., Paschalis, E. P., Fratzl, P. & Klaushofer, K. Bone mineralization density distribution in health and disease. Bone 42, 456–466 (2008). · DOI
  89. Pritchard, J. M. et al. Bone mineralization is elevated and less heterogeneous in adults with type 2 diabetes and osteoarthritis compared to controls with osteoarthritis alone. Bone 54, 76–82 (2013). · DOI
  90. Busse, B. et al. Increased calcium content and inhomogeneity of mineralization render bone toughness in osteoporosis: mineralization, morphology and biomechanics of human single trabeculae. Bone 45, 1034–1043 (2009). · DOI
  91. Fratzl-Zelman, N. et al. Mineral particle size in children with osteogenesis imperfecta type I is not increased independently of specific collagen mutations. Bone 60, 122–128 (2014). · DOI
  92. Fratzl-Zelman, N. et al. Unique micro- and nano-scale mineralization pattern of human osteogenesis imperfecta type VI bone. Bone 73, 233–241 (2015). · DOI
  93. Roschger, P. et al. Alendronate increases degree and uniformity of mineralization in cancellous bone and decreases the porosity in cortical bone of osteoporotic women. Bone 29, 185–191 (2001). · DOI
  94. Koehne, T. et al. Trends in trabecular architecture and bone mineral density distribution in 152 individuals aged 30–90 years. Bone 66, 31–38 (2014). · DOI
  95. Stewart, T. L. et al. Association of COLIA1 Sp1 alleles with defective bone nodule formation in vitro and abnormal bone mineralization in vivo. Calcif. Tissue Int. 77, 113–118 (2005). · DOI
  96. Mahamid, J. et al. Mapping amorphous calcium phosphate transformation into crystalline mineral from the cell to the bone in zebrafish fin rays. Proc. Natl Acad. Sci. USA 107, 6316–6321 (2010). · DOI · Paper page
  97. Kerschnitzki, M. et al. Bone mineralization pathways during the rapid growth of embryonic chicken long bones. J. Struct. Biol. 195, 82–92 (2016). · DOI
  98. Karazisis, D. et al. The role of well-defined nanotopography of titanium implants on osseointegration: cellular and molecular events in vivo. Int. J. Nanomed. 11, 1367–1382 (2016).
  99. Davies, J. E., Ajami, E., Moineddin, R. & Mendes, V. C. The roles of different scale ranges of surface implant topography on the stability of the bone/implant interface. Biomaterials 34, 3535–3546 (2013). · DOI
  100. Davies, J. E., Mendes, V. C., Ko, J. C. & Ajami, E. Topographic scale-range synergy at the functional bone/implant interface. Biomaterials 35, 25–35 (2014). · DOI
  101. Traini, T., Degidi, M., Strocchi, R., Caputi, S. & Piattelli, A. Collagen fiber orientation near dental implants in human bone: do their organization reflect differences in loading? J. Biomed. Mater. Res. B 74B, 538–546 (2005). · DOI
  102. Davies, J. E. Understanding peri-implant endosseous healing. J. Dent. Educ. 67, 932–949 (2003). · DOI
  103. Hoerth, R. M. et al. Mechanical and structural properties of bone in non-critical and critical healing in rat. Acta Biomater. 10, 4009–4019 (2014). · DOI
  104. Pobloth, A.-M. et al. Mechanobiologically optimized 3D titanium-mesh scaffolds enhance bone regeneration in critical segmental defects in sheep. Sci. Transl. Med. 10, eaam8828 (2018). · DOI
  105. Kerschnitzki, M. et al. Poorly ordered bone as an endogenous scaffold for the deposition of highly oriented lamellar tissue in rapidly growing ovine bone. Cells Tissues Organs 194, 119–123 (2011). · DOI
  106. Franchi, M. et al. Early detachment of titanium particles from various different surfaces of endosseous dental implants. Biomaterials 25, 2239–2246 (2004). · DOI
  107. Franchi, M. et al. Biological fixation of endosseous implants. Micron 36, 665–671 (2005). · DOI
  108. Shah, F. A. & Palmquist, A. Evidence that osteocytes in autogenous bone fragments can repair disrupted canalicular networks and connect with osteocytes in de novo formed bone on the fragment surface. Calcif. Tissue Int. 101, 321–327 (2017). · DOI
  109. Sumner, D. R., Bryan, J. M., Urban, R. M. & Kuszak, J. R. Measuring the volume fraction of bone ingrowth: a comparison of three techniques. J. Orthop. Res. 8, 448–452 (1990). · DOI
  110. Shah, F. A., Nilson, B., Brånemark, R., Thomsen, P. & Palmquist, A. The bone–implant interface—nanoscale analysis of clinically retrieved dental implants. Nanomedicine 10, 1729–1737 (2014). · DOI
  111. Stenlund, P. et al. Osseointegration enhancement by Zr doping of Co–Cr–Mo implants fabricated by electron beam melting. Addit. Manuf. 6, 6–15 (2015). · DOI
  112. Omar, O. M. et al. The correlation between gene expression of proinflammatory markers and bone formation during osseointegration with titanium implants. Biomaterials 32, 374–386 (2011). · DOI
  113. Palmquist, A., Emanuelsson, L., Brånemark, R. & Thomsen, P. Biomechanical, histological and ultrastructural analyses of laser micro- and nano-structured titanium implant after 6 months in rabbit. J. Biomed. Mater. Res B 97, 289–298 (2011). · DOI
  114. Palmquist, A. et al. Biomechanical, histological, and ultrastructural analyses of laser micro- and nano-structured titanium alloy implants: a study in rabbit. J. Biomed. Mater. Res. A 92, 1476–1486 (2010). · DOI
  115. Karlsson, J. et al. Ex vivo alendronate localization at the mesoporous titania implant/bone interface. J. Mater. Sci. Mater. Med. 26, 11 (2015). · DOI
  116. Braem, A. et al. Peri- and intra-implant bone response to microporous Ti coatings with surface modification. Acta Biomater. 10, 986–995 (2014). · DOI
  117. Stenlund, P. et al. Bone response to a novel Ti–Ta–Nb–Zr alloy. Acta Biomater. 20, 165–175 (2015). · DOI
  118. Palmquist, A., Windahl, S. H., Norlindh, B., Brånemark, R. & Thomsen, P. Retrieved bone-anchored percutaneous amputation prosthesis showing maintained osseointegration after 11 years—a case report. Acta Orthop. 85, 442–445 (2014). · DOI
  119. Bloebaum, R. D., Koller, K. E., Willie, B. M. & Hofmann, A. A. Does using autograft bone chips achieve consistent bone ingrowth in primary TKA? Clin. Orthop. Relat. Res. 470, 1869–1878 (2012). · DOI
  120. Bobyn, J. D., Stackpool, G. J., Hacking, S. A., Tanzer, M. & Krygier, J. J. Characteristics of bone ingrowth and interface mechanics of a new porous tantalum biomaterial. J. Bone Jt. Surg. Br. 81, 907–914 (1999). · DOI
  121. Hofmann, A. A., Bloebaum, R. D. & Bachus, K. N. Progression of human bone ingrowth into porous-coated implants. Rate bone ingrowth in humans. Acta Orthop. Scand. 68, 161–166 (1997). · DOI
  122. Palmquist, A., Snis, A., Emanuelsson, L., Browne, M. & Thomsen, P. Long-term biocompatibility and osseointegration of electron beam melted, free-form-fabricated solid and porous titanium alloy: experimental studies in sheep. J. Biomater. Appl. 27, 1003–1016 (2013). · DOI
  123. Willie, B. M. et al. Osseointegration into a novel titanium foam implant in the distal femur of a rabbit. J. Biomed. Mater. Res B 92, 479–488 (2010). · DOI
  124. Willie, B. M. et al. Cancellous bone osseointegration is enhanced by in vivo loading. Tissue Eng. Part C 16, 1399–1406 (2010). · DOI
  125. Itälä, A. I., Ylänen, H. O., Ekholm, C., Karlsson, K. H. & Aro, H. T. Pore diameter of more than 100 microm is not requisite for bone ingrowth in rabbits. J. Biomed. Mater. Res. 58, 679–683 (2001). · DOI
  126. Mendes, V. C., Moineddin, R. & Davies, J. E. The effect of discrete calcium phosphate nanocrystals on bone-bonding to titanium surfaces. Biomaterials 28, 4748–4755 (2007). · DOI
  127. Omar, O. et al. In vivo gene expression in response to anodically oxidized versus machined titanium implants. J. Biomed. Mater. Res. A 92, 1552–1566 (2010). · DOI
  128. Luklinska, Z. B. & Bonfield, W. Morphology and ultrastructure of the interface between hydroxyapatite–polyhydroxybutyrate composite implant and bone. J. Mater. Sci. Mater. Med. 8, 379–383 (1997). · DOI
  129. Ma, R. et al. Osseointegration of nanohydroxyapatite- or nano-calcium silicate-incorporated polyetheretherketone bioactive composites in vivo. Int. J. Nanomed. 11, 6023–6033 (2016). · DOI
  130. Merolli, A., Tranquilli Leali, P. & De Santis, E. A back-scattered electron microscopy (BSEM) study of the tight apposition between bone and hydroxyapatite coating. J. Orthop. Trauma 1, 11–16 (2000). · DOI
  131. Stadelmann, V. A., Gauthier, O., Terrier, A., Bouler, J. M. & Pioletti, D. P. Implants delivering bisphosphonate locally increase periprosthetic bone density in an osteoporotic sheep model. A pilot study. Eur. Cells Mater. 16, 10–16 (2008). · DOI
  132. Peter, B. et al. Calcium phosphate drug delivery system: influence of local zoledronate release on bone implant osteointegration. Bone 36, 52–60 (2005). · DOI
  133. Peter, B. et al. Local delivery of bisphosphonate from coated orthopedic implants increases implants mechanical stability in osteoporotic rats. J. Biomed. Mater. Res. A 76A, 133–143 (2006). · DOI
  134. Stadelmann, V. A., Terrier, A., Gauthier, O., Bouler, J. M. & Pioletti, D. P. Prediction of bone density around orthopedic implants delivering bisphosphonate. J. Biomech. 42, 1206–1211 (2009). · DOI
  135. Verron, E. et al. In vivo bone augmentation in an osteoporotic environment using bisphosphonate-loaded calcium deficient apatite. Biomaterials 31, 7776–7784 (2010). · DOI
  136. Boyde, A., Corsi, A., Quarto, R., Cancedda, R. & Bianco, P. Osteoconduction in large macroporous hydroxyapatite ceramic implants: evidence for a complementary integration and disintegration mechanism. Bone 24, 579–589 (1999). · DOI
  137. Elgali, I. et al. Molecular and structural patterns of bone regeneration in surgically created defects containing bone substitutes. Biomaterials 35, 3229–3242 (2014). · DOI
  138. Dias, A. G. et al. In vivo performance of biodegradable calcium phosphate glass ceramics using the rabbit model: histological and SEM observation. J. Biomater. Appl. 20, 253–266 (2006). · DOI
  139. Le Nihouannen, D. et al. Ectopic bone formation by microporous calcium phosphate ceramic particles in sheep muscles. Bone 36, 1086–1093 (2005). · DOI
  140. Le Nihouannen, D. et al. Bone tissue formation in sheep muscles induced by a biphasic calcium phosphate ceramic and fibrin glue composite. J. Mater. Sci. Mater. Med. 19, 667–675 (2008). · DOI
  141. Le Nihouannen, D. et al. Hybrid composites of calcium phosphate granules, fibrin glue, and bone marrow for skeletal repair. J. Biomed. Mater. Res. A 81, 399–408 (2007). · DOI
  142. Bellucci, D., Anesi, A., Salvatori, R., Chiarini, L. & Cannillo, V. A comparative in vivo evaluation of bioactive glasses and bioactive glass-based composites for bone tissue repair. Mater. Sci. Eng. C 79, 286–295 (2017). · DOI
  143. Freeman, C. O. et al. Crystallization modifies osteoconductivity in an apatite-mullite glass-ceramic. J. Mater. Sci. Mater. Med. 14, 985–990 (2003). · DOI
  144. Goodridge, R. D., Wood, D. J., Ohtsuki, C. & Dalgarno, K. W. Biological evaluation of an apatite-mullite glass-ceramic produced via selective laser sintering. Acta Biomater. 3, 221–231 (2007). · DOI
  145. Liu, X., Rahaman, M. N. & Fu, Q. Bone regeneration in strong porous bioactive glass (13-93) scaffolds with an oriented microstructure implanted in rat calvarial defects. Acta Biomater. 9, 4889–4898 (2013). · DOI
  146. Liu, X., Rahaman, M. N., Liu, Y., Bal, B. S. & Bonewald, L. F. Enhanced bone regeneration in rat calvarial defects implanted with surface-modified and BMP-loaded bioactive glass (13-93) scaffolds. Acta Biomater. 9, 7506–7517 (2013). · DOI
  147. Müller-Mai, C., Voigt, C., de Almeida Reis, S. R., Herbst, H. & Gross, U. M. Substitution of natural coral by cortical bone and bone marrow in the rat femur. J. Mater. Sci. Mater. Med. 7, 479–488 (1996). · DOI
  148. Liao, H., Mutvei, H., Sjöström, M., Hammarström, L. & Li, J. Tissue responses to natural aragonite (Margaritifera shell) implants in vivo. Biomaterials 21, 457–468 (2000). · DOI
  149. Pascaretti-Grizon, F. et al. The interface between nacre and bone after implantation in the sheep: a nanotomographic and Raman study. J. Raman Spectrosc. 45, 558–564 (2014). · DOI
  150. Cipitria, A. et al. Porous scaffold architecture guides tissue formation. J. Bone Miner. Res. 27, 1275–1288 (2012). · DOI
  151. Paris, M. et al. Scaffold curvature-mediated novel biomineralization process originates a continuous soft tissue-to-bone interface. Acta Biomater. 60, 64–80 (2017). · DOI
  152. Woodruff, M. A., Lange, C., Chen, F., Fratzl, P. & Hutmacher, D. W. Nano- to macroscale remodeling of functional tissue-engineered bone. Adv. Healthc. Mater. 2, 546–551 (2013). · DOI
  153. Cipitria, A. et al. BMP delivery complements the guiding effect of scaffold architecture without altering bone microstructure in critical-sized long bone defects: a multiscale analysis. Acta Biomater. 23, 282–294 (2015). · DOI
  154. Reichert, J. C. et al. A tissue engineering solution for segmental defect regeneration in load-bearing long bones. Sci. Transl. Med. 4, 141ra193 (2012). · DOI
  155. Shah, F. A., Stenlund, P., Martinelli, A., Thomsen, P. & Palmquist, A. Direct communication between osteocytes and acid-etched titanium implants with a sub-micron topography. J. Mater. Sci. Mater. Med. 27, 167 (2016). · DOI
  156. Du, Z., Ivanovski, S., Hamlet, S. M., Feng, J. Q. & Xiao, Y. The ultrastructural relationship between osteocytes and dental implants following osseointegration. Clin. Implant Dent. Relat. Res. 18, 270–280 (2016). · DOI
  157. Shah, F. A., Wang, X., Thomsen, P., Grandfield, K. & Palmquist, A. High-resolution visualization of the osteocyte lacuno-canalicular network juxtaposed to the surface of nanotextured titanium implants in human. ACS Biomater. Sci. Eng. 1, 305–313 (2015). · DOI
  158. Gorustovich, A. A. Imaging resin-cast osteocyte lacuno-canalicular system at bone-bioactive glass interface by scanning electron microscopy. Microsc. Microanal. 16, 132–136 (2010). · DOI
  159. Åkesson, K., Grynpas, M. D., Hancock, R. G., Odselius, R. & Obrant, K. J. Energy-dispersive X-ray microanalysis of the bone mineral content in human trabecular bone: a comparison with ICPES and neutron activation analysis. Calcif. Tissue Int. 55, 236–239 (1994). · DOI
  160. Obrant, K. J. & Odselius, R. Electron microprobe investigation of calcium and phosphorus concentration in human bone trabeculae—both normal and in posttraumatic osteopenia. Calcif. Tissue Int. 37, 117–120 (1985). · DOI
  161. Wierzchos, J. et al. Advances in the ultrastructural study of the implant–bone interface by backscattered electron imaging. Micron 39, 1363–1370 (2008). · DOI
  162. Gupta, H. S. et al. Two different correlations between nanoindentation modulus and mineral content in the bone–cartilage interface. J. Struct. Biol. 149, 138–148 (2005). · DOI
  163. Milovanovic, P. et al. The formation of calcified nanospherites during micropetrosis represents a unique mineralization mechanism in aged human bone. Small 13, 1602215 (2017). · DOI
  164. Brüel, A. et al. Strontium is incorporated into the fracture callus but does not influence the mechanical strength of healing rat fractures. Calcif. Tissue Int. 88, 142–152 (2011). · DOI
  165. Ballo, A. M. et al. Bone tissue reactions to biomimetic ion-substituted apatite surfaces on titanium implants. J. R. Soc. Interface 9, 1615–1624 (2012). · DOI
  166. Ballo, A. M. et al. Osseointegration of fiber-reinforced composite implants: histological and ultrastructural observations. Dent. Mater. 30, e384–e395 (2014). · DOI
  167. Thomas, A. et al. Direct observation of bone coherence with dental implants. J. Eur. Ceram. Soc. 32, 2607–2612 (2012). · DOI
  168. Chen, P.-Y. & McKittrick, J. Compressive mechanical properties of demineralized and deproteinized cancellous bone. J. Mech. Behav. Biomed. Mater. 4, 961–973 (2011). · DOI
  169. Congiu, T., Pazzaglia, U. E., Basso, P. & Quacci, D. Chemical etching in processing cortical bone specimens for scanning electron microscopy. Microsc. Res. Tech. 77, 653–660 (2014). · DOI
  170. Chen, P.-Y., Toroian, D., Price, P. A. & McKittrick, J. Minerals form a continuum phase in mature cancellous bone. Calcif. Tissue Int. 88, 351–361 (2011). · DOI
  171. Yamamoto, T. et al. Structure and formation of the twisted plywood pattern of collagen fibrils in rat lamellar bone. J. Electron Microsc. 61, 113–121 (2012).
  172. Ardizzoni, A. Osteocyte lacunar size-lamellar thickness relationships in human secondary osteons. Bone 28, 215–219 (2001). · DOI
  173. Pazzaglia, U. E., Congiu, T., Marchese, M., Spagnuolo, F. & Quacci, D. Morphometry and patterns of lamellar bone in human Haversian systems. Anat. Rec. 295, 1421–1429 (2012). · DOI
  174. Congiu, T. & Pazzaglia, U. E. The sealed osteons of cortical diaphyseal bone. Early observations revisited with scanning electron microscopy. Anat. Rec. 294, 193–198 (2011). · DOI
  175. Mahamid, J., Sharir, A., Addadi, L. & Weiner, S. Amorphous calcium phosphate is a major component of the forming fin bones of zebrafish: Indications for an amorphous precursor phase. Proc. Natl Acad. Sci. USA 105, 12748–12753 (2008). · DOI · Paper page
  176. Midura, R. J. et al. Calcospherulites isolated from the mineralization front of bone induce the mineralization of type I collagen. Bone 41, 1005–1016 (2007). · DOI
  177. Midura, R. J., Vasanji, A., Su, X., Midura, S. B. & Gorski, J. P. Isolation of calcospherulites from the mineralization front of bone. Cells Tissues Organs 189, 75–79 (2009). · DOI
  178. Pazzaglia, U. E. et al. The application of heat-deproteinization to the morphological study of cortical bone: a contribution to the knowledge of the osteonal structure. Microsc. Res. Tech. 79, 691–699 (2016). · DOI
  179. Holden, J. L., Phakey, P. P. & Clement, J. G. Scanning electron microscope observations of heat-treated human bone. Forensic Sci. Int. 74, 29–45 (1995). · DOI
  180. Ejiri, S. & Ozawa, H. Scanning electron microscopic observations of rat tibia using the HCl-collagenase method. Arch. Histol. Jpn. 45, 399–404 (1982). · DOI
  181. Abe, K., Hashizume, H. & Ushiki, T. An EDTA-KOH method to expose bone cells for scanning electron microscopy. J. Electron Microsc. 41, 113–115 (1992).
  182. Pazzaglia, U. E. & Congiu, T. The cast imaging of the osteon lacunar-canalicular system and the implications with functional models of intracanalicular flow. J. Anat. 222, 193–202 (2013). · DOI
  183. Bohner, M. et al. Characterization and distribution of mechanically competent mineralized tissue in micropores of β-tricalcium phosphate bone substitutes. Mater. Today 20, 106–115 (2017). · DOI
  184. Du, Z., Xiao, Y., Hashimi, S., Hamlet, S. M. & Ivanovski, S. The effects of implant topography on osseointegration under estrogen deficiency induced osteoporotic conditions: Histomorphometric, transcriptional and ultrastructural analysis. Acta Biomater. 42, 351–363 (2016). · DOI
  185. Lin, T. C., Su, C. Y. & Chang, C. S. Stereomorphologic observation of bone tissue response to hydroxyapatite using SEM with the EDTA-KOH method. J. Biomed. Mater. Res. 36, 91–97 (1997). · DOI
  186. Gung, Y. W., Cheng, C. K. & Su, C. Y. A stereomorphologic study of bone matrix apposition in HA-implanted cavities observed with SEM, being prepared by a microvascular cast and freeze-fracture method. Med. Eng. Phys. 25, 565–571 (2003). · DOI
  187. Chang, C. S., Su, C. Y. & Lin, T. C. Scanning electron microscopy observation of vascularization around hydroxyapatite using vascular corrosion casts. J. Biomed. Mater. Res. 48, 411–416 (1999). · DOI
  188. Martinez-Maza, C., Rosas, A., García-Vargas, S., Estalrrich, A. & de la Rasilla, M. Bone remodelling in Neanderthal mandibles from the El Sidrón site (Asturias, Spain). Biol. Lett. 7, 593–596 (2011). · DOI
  189. Mansilla, J. et al. On porosity of archeological bones II. Textural characterization of Mesoamerican human bones. Palaeogeogr. Palaeoclim. Palaeoecol. 414, 493–499 (2014). · DOI
  190. Bosch, P. et al. On porosity of archeological bones I—textural characterization of pathological Spanish medieval human bones. Palaeogeogr. Palaeoclim. Palaeoecol. 414, 486–492 (2014). · DOI
  191. Brickley, M. & Ives, R. Skeletal manifestations of infantile scurvy. Am. J. Phys. Anthropol. 129, 163–172 (2006). · DOI
  192. Mays, S., Brickley, M. & Ives, R. Skeletal evidence for hyperparathyroidism in a 19th century child with rickets. Int. J. Osteoarchaeol. 17, 73–81 (2007). · DOI
  193. Brickley, M., Mays, S. & Ives, R. An investigation of skeletal indicators of vitamin D deficiency in adults: effective markers for interpreting past living conditions and pollution levels in 18th and 19th century Birmingham, England. Am. J. Phys. Anthropol. 132, 67–79 (2007). · DOI
  194. Bell, L. S. in Forensic Microscopy for Skeletal Tissues, Vol. 915, Ch. 911, 173–190 (ed. Bell, L. S.) (Humana Press Totowa, NJ, 2012). · DOI
  195. Turner-Walker, G. & Jans, M. Reconstructing taphonomic histories using histological analysis. Palaeogeogr. Palaeoclim. Palaeoecol. 266, 227–235 (2008). · DOI
  196. Fernández-Jalvo, Y. et al. Early bone diagenesis in temperate environments: Part I: Surface features and histology. Palaeogeogr. Palaeoclim. Palaeoecol. 288, 62–81 (2010). · DOI
  197. Keenan, S. W. & Engel, A. S. Reconstructing diagenetic conditions of bone at the Gray Fossil Site, Tennessee, USA. Palaeogeogr. Palaeoclim. Palaeoecol. 471, 48–57 (2017). · DOI
  198. Dal Sasso, G., Maritan, L., Usai, D., Angelini, I. & Artioli, G. Bone diagenesis at the micro-scale: Bone alteration patterns during multiple burial phases at Al Khiday (Khartoum, Sudan) between the Early Holocene and the II century AD. Palaeogeogr. Palaeoclim. Palaeoecol. 416, 30–42 (2014). · DOI
  199. Hollund, H. I., Blank, M. & Sjögren, K. G. Dead and buried? Variation in post-mortem histories revealed through histotaphonomic characterisation of human bone from megalithic graves in Sweden. PLoS One 13, e0204662 (2018). · DOI
  200. Tjelldén, A. K. E., Kristiansen, S. M., Birkedal, H. & Jans, M. M. E. The pattern of human bone dissolution—a histological study of Iron Age warriors from a Danish wetland site. Int J. Osteoarchaeol. 28, 407–418 (2018). · DOI

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Metadata

Authors
Shah, F. A., Ruscsák, K., & Palmquist, A.
Venue
Bone Research
Publisher
Springer Science and Business Media LLC
Published
2019-05-22
Volume
7
Issue
1
ISSN
2095-6231
DOI
10.1038/s41413-019-0053-z
PMC
PMC6531483
Cited by
115
References
3
Language
en
Licence

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

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How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: 50 years of scanning electron microscopy of bone—a comprehensive overview of the important discoveries made and insights gained into bone material properties in health, disease, and taphonomy. https://app.cochat.ai/papers/10.1038/s41413-019-0053-z
The paper
Shah, F. A., Ruscsák, K., & Palmquist, A. (2019). 50 years of scanning electron microscopy of bone—a comprehensive overview of the important discoveries made and insights gained into bone material properties in health, disease, and taphonomy. Bone Research. https://doi.org/10.1038/s41413-019-0053-z