The ultrastructure of bone as revealed in electron microscopy of ion-milled sections

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Schwarcz, Henry P.¹

The ultrastructure of bone as revealed in electron microscopy of ion-milled sections

Semin Cell Dev Biol. October 2015;46:44-50

©2015 Elsevier Ltd. All rights reserved.

Affiliations

¹School of Geography and Earth Sciences, McMaster University, Hamilton, ON, Canada
Abstract

Mineral makes up more than half the volume of bone, but its spatial and structural relationship to collagen and other proteins is still a matter of debate. Due to the nanometer-size of bone crystals this matter can be resolved only with transmission electron microscope (TEM) images. Using sections cut with an ultramicrotome, previous investigators determined most mineral lies in the 40 nm wide gap zone in collagen fibrils. Using less invasive sectioning methods (ion milling and focused ion beam [FIB]) reveals that most mineral is extrafibrillar, occurring in the form of mineral lamellae, polycrystalline plates 300 nm or more long, packed around collagen fibrils in stacks of four or more lamellae <1 nm apart. While Ca and P also occur in the gap zone, they do not appear to be in the form of well-crystallized apatite. This new model for bone ultrastructure resolves outstanding problems presented by the previous model.
Links

DOI: 10.1016/j.semcdb.2015.06.008

PubMed: 26165821

WoS: 000366137400007
History

Received: 2015-03-4

Revised: 2015-05-22

Accepted: 2015-06-28

Online: 2015-07-9

Cited Works (19)

Year	Entry
1999	Rinnerthaler S, Roschger P, Jakob HF, Nader A, Klaushofer K, Fratzl P. Scanning small angle X-ray scattering analysis of human bone sections. Calcif Tiss Int. May 1999;64(5):422-429.
1986	Weiner S, Price PA. Disaggregation of bone into crystals. Calcif Tiss Int. November 1986;39(6):365-375.
2010	Nudelman F, Pieterse K, George A, Bomans PHH, Friedrich H, Brylka LJ, Hilbers PAJ, de With G, Sommerdijk NAJM. The role of collagen in bone apatite formation in the presence of hydroxyapatite nucleation inhibitors. Nat Mater. December 2010;9(12):1004-1009.
2014	Schwarcz HP, McNally EA, Botton GA. Dark-field transmission electron microscopy of cortical bone reveals details of extrafibrillar crystals. J Struct Biol. December 2014;188(3):240-248.
2003	Su X, Sun K, Cui FZ, Landis WJ. Organization of apatite crystals in human woven bone. Bone. February 2003;32(2):150-162.
2006	Orgel JPRO, Irving TC, Miller A, Wess TJ. Microfibrillar structure of type I collagen in situ. Proc Natl Acad Sci USA. June 13, 2006;103(24):9001-9005.
2012	McNally EA, Schwarcz HP, Botton GA, Arsenault AL. A model for the ultrastructure of bone based on electron microscopy of ion-milled sections. PLoS One. January 17, 2012;7(1):e29258.
1991	Weiner S, Arad T, Traub W. Crystal organization in rat bone lamellae. FEBS Lett. July 1991;285(1):49-54.
1993	Landis WJ, Song MJ, Leith A, McEwen L, McEwen BF. Mineral and organic matrix interaction in normally calcifying tendon visualized in three dimensions by high-voltage electron microscopic tomography and graphic image reconstruction. J Struct Biol. January 1993;110(1):39-54.
2012	Alexander B, Daulton TL, Genin GM, Lipner J, Pasteris JD, Wopenka B, Thomopoulos S. The nanometre-scale physiology of bone: steric modelling and scanning transmission electron microscopy of collagen-mineral structure. J R Soc Interface. August 7, 2012;9(73):1774-1786.
2002	Benezra Rosen V, Hobbs LW, Spector M. The ultrastructure of anorganic bovine bone and selected synthetic hyroxyapatites used as bone graft substitute materials. Biomaterials. February 2002;23(3):921-928.
1999	Wenk H-R, Heidelbach F. Crystal alignment of carbonated apatite in bone and calcified tendon: results from quantitative texture analysis. Bone. April 1999;24(4):361-369.
2000	Jäger I, Fratzl P. Mineralized collagen fibrils: a mechanical model with a staggered arrangement of mineral particles. Biophys J. October 2000;79(4):1737-1746.
2007	Olszta MJ, Cheng X, Jee SS, Kumar R, Kim Y-Y, Kaufman MJ, Douglas EP, Gower LB. Bone structure and formation: a new perspective. Mater Sci Eng R Rep. November 28, 2007;58(3-5):77-116.
2011	Chen P-Y, Toroian D, Price PA, McKittrick J. Minerals form a continuum phase in mature cancellous bone. Calcif Tiss Int. May 2011;88(5):351-361.
2001	Eppell SJ, Tong W, Katz JL, Kuhn L, Glimcher MJ. Shape and size of isolated bone mineralites measured using atomic force microscopy. J Orthop Res. November 2001;19(6):1027-1034.
2014	Reznikov N, Shahar R, Weiner S. Bone hierarchical structure in three dimensions. Acta Biomater. September 2014;10(8):3815-3826.
2003	Rubin MA, Jasiuk I, Taylor J, Rubin J, Ganey T, Apkarian RP. TEM analysis of the nanostructure of normal and osteoporotic human trabecular bone. Bone. March 2003;32(3):270-282.
1996	McKee MD, Nanci A. Osteopontin at mineralized tissue interfaces in bone, teeth, and osseointegrated implants: ultrastructural distribution and implications for mineralized tissue formation, turnover, and repair. Microsc Res Tech. February 1996;33(2):141-164.

Cited By (12)

Year	Entry
2020	Schwarcz HP, Binkley DM, Luo L, Grandfield K. A search for apatite crystals in the gap zone of collagen fibrils in bone using dark-field illumination. Bone. June 2020;135:115304.
2018	Grandfield K, Vuong V, Schwarcz HP. Ultrastructure of bone: hierarchical features from nanometer to micrometer scale revealed in focused ion beam sections in the TEM. Calcif Tiss Int. December 2018;103(6):606-616.
2020	Buss DJ, Reznikov N, McKee MD. Crossfibrillar mineral tessellation in normal and Hyp mouse bone as revealed by 3D FIB-SEM microscopy. J Struct Biol. November 1, 2020;212(2):107603.
2020	Binkley DM, Deering J, Yuan H, Gourrier A, Grandfield K. Ellipsoidal mesoscale mineralization pattern in human cortical bone revealed in 3D by plasma focused ion beam serial sectioning. J Struct Biol. November 1, 2020;212(2):107615.
2019	Bian T, Zhao K, Meng Q, Tang Y, Jiao H, Luo J. The construction and performance of multi-level hierarchical hydroxyapatite (HA)/collagen composite implant based on biomimetic bone Haversian motif. Mater Des. January 15, 2019;162:60-69.
2018	Reznikov N, Bilton M, Lari L, Stevens MM, Kröger R. Fractal-like hierarchical organization of bone begins at the nanoscale. Science. May 4, 2018;360(6388):eaao2189.
2023	Buss DJ. Mineralization of Biological Fiber Systems [PhD thesis]. McGill University; July 2023.
2017	Tedesco J. Multi-Scale Characterization of Biomaterials Using Complementary X-Ray and Electron Imaging Techniques [Master's thesis]. Hamilton, ON: McMaster University; September 2017.
2017	Wang X. Multi-Dimensional Characterization of Bone and Bone-Implant Interfaces [PhD thesis]. Hamilton, ON: McMaster University; December 2017.
2019	Binkley DM. Electron and Ion Beam Imaging of Human Bone Structure Across the Nano- and Mesoscale [Master's thesis]. Hamilton, ON: McMaster University; June 2019.
2019	Lee BEJ. Characterization Strategies for Bone Ultrastructure and Bone-Cell Interfacing Materials [PhD thesis]. Hamilton, ON: McMaster University; November 2019.
2019	Strakhov I. The Nanoscale Structure of Human Female Osteoporotic Bone Investigated by Transmission Electron Microscopy [Master's thesis]. Hamilton, ON: McMaster University; September 2019.