Multiscale Quantitative Imaging of Human Femoral Heads Using X-Ray Microtomography

Clinical diagnostic tools provide limited information on the underlying structural and mechanical properties of bone-tissue affected by degenerative and bone metabolic diseases. In-vivo bone failure studies provide limited information due to constraints such as X-ray dosage, cost and various other practicalities. In-vitro studies are thus required to enhance understanding of this phenomenon. The aims of this study were to use quantitative high-definition X-ray Micro-Tomography (XMT) to assess factors contributing to pathological and non-pathological bone failure and repair in relation to the mechanics of whole human femoral heads.

XMT images of one normal and six pathological femoral heads were collected at 26 – 8.8 μm voxel resolution and evaluated to determine structural features; bone mineral concentration (BMC); and using image analysis, identify microcallus formations. In addition, in-vitro compression tests were carried out on specimens taken from regions with different anatomical loading. Bone quality was then related to the anatomical loading and BMC.

Results from non-pathological tissue where used to establish a baseline for measurements of structural features. Microcallus formations where identified and used as markers to map the occurrence of bone damage. In osteoarthritic (OA) heads, the damage was found to be concentrated in localised clusters. Conversely, in the osteoporotic head damage was distributed homogeneously throughout the entire specimen. No significant difference in the BMC was observed, however there was a significant difference in the bone quality values between the non-pathological and pathological heads, and also between the pathologies. In-vitro mechanical testing revealed a difference in the mechanical properties of OA trabecular bone in relation to bone quality measurements but the samples exhibited no significant correlation to anatomical loading.

X-ray Ultra Microscopy (XuM) at 200nm and 775nm voxel resolution was used to investigate the nano-morphology of individual trabeculae. The XuM images showed differences in bone structure and fewer osteocyte lacunae present close to fracture site. XuM also identified micro-cracks within trabeculae that were encased by microcallus formations.

The application of novel quantitative high definition X-ray imaging to clinically relevant tissue at multiple length scales has provided new metrological data on the distribution of damage within pathological tissue. Insight into the vulnerability of diseased tissue to damage could ultimately lead to improved diagnosis from clinical radiographs.

Year	Entry
1985	Gibson LJ. The mechanical behaviour of cancellous bone. J Biomech. 1985;18(5):317-328.
1973	Vernon-Roberts B, Pirie CJ. Healing trabecular microfractures in the bodies of lumbar vertebrae. Ann Rheum Dis. September 1973;32(5):406-412.
2002	Currey JD. Bones: Structure and Mechanics. Princeton, NJ: Princeton University Press; 2002.
2000	Timlin JA, Carden A, Morris MD, Rajachar RM, Kohn DH. Raman spectroscopic imaging markers for fatigue-related microdamage in bovine bone. Anal Chem. May 15, 2000;72(10):2229-2236.
1983	Brown TD, Shaw DT. In vitro contact stress distributions in the natural human hip. J Biomech. 1983;16(6):373-384.
1995	Landis WJ. The strength of a calcified tissue depends in part on the molecular structure and organization of its constituent mineral crystals in their organic matrix. Bone. May 1995;16(5):533-544.
1999	Zysset PK, Guo XE, Hoffler CE, Moore KE, Goldstein SA. Elastic modulus and hardness of cortical and trabecular bone lamellae measured by nanoindentation in the human femur. J Biomech. October 1999;32(10):1005-1012.
1965	Sedlin ED. A rheologic model for cortical bone: a study of physical properties of human femoral samples. Acta Orthop Scand. 1965;(suppl 83):1-77.
1991	Ciarelli MJ, Goldstein SA, Kuhn JL, Cody DD, Brown MB. Evaluation of orthogonal mechanical properties and density of human trabecular bone from the major metaphyseal regions with materials testing and computed tomography. J Orthop Res. May 1991;9(5):674-682.
1985	Beaupre GS, Hayes WC. Finite element analysis of a three-dimensional open-celled model for trabecular bone. J Biomech Eng. August 1985;107(3):249-256.
1997	Hildebrand T, Rüegsegger P. Quantification of bone microarchitecture with the structure model index. Comput Methods Prog Biomed. 1997;1(1):15-23.
2007	Perilli E, Baleani M, Öhman C, Baruffaldi F, Viceconti M. Structural parameters and mechanical strength of cancellous bone in the femoral head in osteoarthritis do not depend on age. Bone. November 2007;41(5):760-768.
2003	Ferguson VL, Bushby AJ, Boyde A. Nanomechanical properties and mineral concentration in articular calcified cartilage and subchondral bone. J Anat. August 2003;203(2):191-202.
1983	Parfitt AM, Mathews CH, Villanueva AR, Kleerekoper M, Frame B, Rao DS. Relationships between surface, volume, and thickness of iliac trabecular bone in aging and in osteoporosis: implications for the microanatomic and cellular mechanisms of bone loss. J Clin Invest. October 1983;72(4):1396-1409.
1995	Schaffler MB, Choi K, Milgrom C. Aging and matrix microdamage accumulation in human compact bone. Bone. December 1995;17(6):521-525.
1997	Gibson LJ, Ashby MF. Cellular Solids: Structure and Properties. 2nd ed. Cambridge, UK: Cambridge University Press; 1997. Cambridge Solid State Science Series.
2003	Boyde A. The real response of bone to exercise. J Anat. August 2003;203(2):173-189.
1987	Parfitt AM, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR. Bone histomorphometry: standardization of nomenclature, symbols, and units: report of the ASBMR histomorphometry nomenclature committee. J Bone Miner Res. December 1987;2(6):595-610.
2006	Stauber M, Rapillard L, van Lenthe GH, Zysset P, Müller R. Importance of individual rods and plates in the assessment of bone quality and their contribution to bone stiffness. J Bone Miner Res. April 2006;21(4):586-595.
1996	Wenzel TE, Schaffler MB, Fyhrie DP. In vivo trabecular microcracks in human vertebral bone. Bone. August 1996;19(2):89-95.
1993	Fazzalari NL. Trabecular microfracture. Calcif Tiss Int. February 1993;53(suppl 1):S143-S147.
1998	Bentolila V, Boyce TM, Fyhrie DP, Drumb R, Skerry TM, Schaffler MB. Intracortical remodeling in adult rat long bones after fatigue loading. Bone. September 1998;23(3):275-281.
1998	Fazzalari NL, Forwood MR, Smith K, Manthey BA, Herreen P. Assessment of cancellous bone quality in severe osteoarthrosis: bone mineral density, mechanics, and microdamage. Bone. 1998;22(4):381-388.
2006	Stauber M, Müller R. Age-related changes in trabecular bone microstructures: global and local morphometry. Osteoporos Int. April 2006;17(4):616-626.
1990	Mosekilde L. Consequences of the remodelling process for vertebral trabecular bone structure: a scanning electron microscopy study (uncoupling of unloaded structures). Bone Miner. July 1990;10(1):13-35.
1990	Jensen KS, Mosekilde L, Mosekilde L. A model of vertebral trabecular bone architecture and its mechanical properties. Bone. 1990;11(6):417-423.
1998	Mosekilde L. The effect of modelling and remodelling on human vertebral body architecture. Technol Health Care. December 1998;6(5-6):287-297.
2001	Jee WSS. Integrated bone tissue physiology: anatomy and physiology. In: Cowin SC, ed. Bone Mechanics Handbook. 2nd ed. Boca Raton, FL: CRC Press; 2001:chap 1.
1997	Odgaard A. Three-dimensional methods for quantification of cancellous bone architecture. Bone. 1997;20(4):315-328.
1993	Odgaard A, Gundersen HJG. Quantification of connectivity in cancellous bone, with special emphasis on 3-D reconstructions. Bone. March–April 1993;14(2):173-182.
2001	Cowin SC, ed. Bone Mechanics Handbook. 2nd ed. Boca Raton, FL: CRC Press; 2001.
1993	Goldstein SA, Goulet R, McCubbrey D. Measurement and significance of three-dimensional architecture to the mechanical integrity of trabecular bone. Calcif Tiss Int. February 1993;53(suppl 1):S127-S133.
1994	Zioupos P, Currey JD, Sedman AJ. An examination of the micromechanics of failure of bone and antler by acoustic emission tests and laser scanning confocal microscopy. Med Eng Phys. May 1994;16(3):203-212.
2006	Bembey AK, Bushby AJ, Boyde A, Ferguson VL, Oyen ML. Hydration effects on the micro-mechanical properties of bone. J Mater Res. August 2006;21(8):1962-1968.
2003	Martin RB. Fatigue microdamage as an essential element of bone mechanics and biology. Calcif Tiss Int. August 2003;73(2):101-107.
1986	Mosekilde L, Mosekilde L. Normal vertebral body size and compressive strength: relations to age and to vertebral and iliac trabecular bone compressive strength. Bone. 1986;7(3):207-212.
2001	Turner CH, Burr DB. Experimental techniques for bone mechanics. In: Cowin SC, ed. Bone Mechanics Handbook. 2nd ed. Boca Raton, FL: CRC Press; 2001:chap 7.
1992	Blackburn J, Hodgskinson R, Currey JD, Mason JE. Mechanical properties of microcallus in human cancellous bone. J Orthop Res. March 1992;10(2):237-246.
1997	Mori S, Harruff R, Ambrosius W, Burr DB. Trabecular bone volume and microdamage accumulation in the femoral heads of women with and without femoral neck fractures. Bone. December 1997;21(6):521-526.
2003	Currey JD. The many adaptations of bone. J Biomech. 2003;36(10):1487-1495.
1971	Whitehouse WJ, Dyson ED, Jackson CK. The scanning electron microscope in studies of trabecular bone from a human vertebral body. J Anat. April 1971;108(3):481-496.
1998	Müller R, Van Campenhout H, Van Damme B, Van der Perre G, Dequeker J, Hildebrand T, Rüegsegger P. Morphometric analysis of human bone biopsies: a quantitative structural comparison of histological sections and micro-computed tomography. Bone. July 1998;23(1):59-66.
1987	Mosekilde L, Mosekilde L, Danielsen CC. Biomechanical competence of vertebral trabecular bone in relation to ash density and age in normal individuals. Bone. 1987;8(2):79-85.
1985	Mosekilde L, Viidik A, Mosekilde L. Correlation between the compressive strength of iliac and vertebral trabecular bone in normal individuals. Bone. 1985;6(5):291-295.
1989	Mosekilde L. Sex differences in age-related loss of vertebral trabecular bone mass and structure—biomechanical consequences. Bone. 1989;10(6):425-432.
2007	Wang X, Masse DB, Leng H, Hess KP, Ross RD, Roeder RK, Niebur GL. Detection of trabecular bone microdamage by micro-computed tomography. J Biomech. 2007;40(15):3397-3403.
1987	Goldstein SA. The mechanical properties of trabecular bone: dependence on anatomic location and function. J Biomech. 1987;20(11-12):1055-1061.
2004	Nägele E, Kuhn V, Vogt H, Link TM, Müller R, Lochmüller E-M, Eckstein F. Technical considerations for microstructural analysis of human trabecular bone from specimens excised from various skeletal sites. Calcif Tiss Int. 2004;75(1):15-22.
2006	Diab T, Condo KW, Burr DB, Vashishth D. Age-related change in the damage morphology of human cortical bone and its role in bone fragility. Bone. March 2006;38(3):427-431.
2007	Diab T, Vashishth D. Morphology, localization and accumulation of in vivo microdamage in human cortical bone. Bone. March 2007;40(3):612-618.
2005	Diab T, Vashishth D. Effects of damage morphology on cortical bone fragility. Bone. July 2005;37(1):96-102.
1970	Galante J, Rostoker W, Ray RD. Physical properties of trabecular bone. Calcif Tiss Res. 1970;5(1):236-246.
2005	Shefelbine SJ, Simon U, Claes L, Gold A, Gabet Y, Bab I, Müller R, Augat P. Prediction of fracture callus mechanical properties using micro-CT images and voxel-based finite element analysis. Bone. March 2005;36(3):480-488.
2003	Van Rietbergen B, Huiskes R, Eckstein F, Rüegsegger P. Trabecular bone tissue strains in the healthy and osteoporotic human femur. J Bone Miner Res. October 2003;18(10):1781-1788.
1997	Hildebrand T, Rüegsegger P. A new method for the model-independent assessment of thickness in three-dimensional images. J Microsc (Oxford). 1997;185(1):67-75.
1997	McCalden RW, McGeough JA, Court-Brown CM. Age-related changes in the compressive strength of cancellous bone: the relative importance of changes in density and trabecular architecture. J Bone Joint Surg. March 1997;79A(3):421-427.
1992	Linde F, Hvid I, Madsen F. The effect of specimen geometry on the mechanical behaviour of trabecular bone specimens. J Biomech. 1992;25(4):359-368.
1991	Mellish RWE, Ferguson‐Pell MW, Cochran GVB, Lindsay R, Dempster DW. A new manual method for assessing two‐dimensional cancellous bone structure: comparison between iliac crest and lumbar vertebra. J Bone Miner Res. July 1991;6(7):689-696.
2006	Robling AG, Castillo AB, Turner CH. Biomechanical and molecular regulation of bone remodeling. Annu Rev Biomed Eng. 2006;8:455-498.
1983	Stone JL, Beaupre GS, Hayes WC. Multiaxial strength characteristics of trabecular bone. J Biomech. 1983;16(9):743-752.
1973	Pugh JW, Rose RM, Radin EL. A structural model for the mechanical behavior of trabecular bone. J Biomech. 1973;6(6):657-670.
1987	Lorensen WE, Cline HE. Marching cubes: a high resolution 3D surface construction algorithm. Comput Graph. 1987;21(4):163-169.
1960	Frost HM. Presence of microscopic cracks in vivo in bone. Henry Ford Hosp Med Bull. 1960;8(1):25-35.
1996	Ford CM, Keaveny TM. The dependence of shear failure properties of trabecular bone on apparent density and trabecular orientation. J Biomech. 1996;29(10):1309-1317.
2000	Vashishth D, Verborgt O, Divine G, Schaffler MB, Fyhrie DP. Decline in osteocyte lacunar density in human cortical bone is associated with accumulation of microcracks with age. Bone. April 2000;26(4):375-380.
1976	Hayes WC, Carter DR. Postyield behavior of subchondral trabecular bone. J Biomed Mater Res. July 1976;10(4):537-544.
2010	Doube M, Kłosowski MM, Arganda-Carreras I, Cordelières FP, Dougherty RP, Jackson JS, Schmid B, Hutchinson JR, Shefelbine SJ. BoneJ: free and extensible bone image analysis in ImageJ. Bone. December 2010;47(6):1076-1079.
2008	Chen H, Shoumura S, Emura S, Bunai Y. Regional variations of vertebral trabecular bone microstructure with age and gender. Osteoporos Int. October 2008;19(10):1473-1483.
1995	Hahn M, Vogel M, Amling M, Ritzel H, Delling G. Microcallus formations of the cancellous bone: a quantitative analysis of the human spine. J Bone Miner Res. September 1995;10(9):1410-1416.
1977	Carter DR, Hayes WC. The compressive behavior of bone as a two-phase porous structure. J Bone Joint Surg. 1977;59A(7):954-962.
1984	Currey JD. Mechanical Adaptations of Bone. Princeton, NJ: Princeton University Press; 1984.
1987	Reid SA, 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. February 1987;2(1):13-22.
1994	Keaveny TM, Wachtel EF, Guo XE, Hayes WC. Mechanical behavior of damaged trabecular bone. J Biomech. November 1994;27(11):1309-1318.
1957	Kellgren JH, Lawrence JS. Radiological assessment of osteo-arthrosis. Ann Rheum Dis. December 1957;16(4):494-502.
1990	Sharp DJ, Tanner KE, Bonfield W. Measurement of the density of trabecular bone. J Biomech. 1990;23(8):853-857.
1994	Ziv V, Weiner S. Bone crystal sizes: a comparison of transmission electron microscopic and x-ray diffraction line width broadening techniques. Connect Tissue Res. 1994;30(3):165-175.
1984	Feldkamp LA, Davis LC, Kress JW. Practical cone-beam algorithm. J Opt Soc Am. 1984;A1(6):612-619.
2001	Kaufman JJ, Siffert RS. Noninvasive measurement of bone integrity. In: Cowin SC, ed. Bone Mechanics Handbook. 2nd ed. Boca Raton, FL: CRC Press; 2001:chap 34.
1997	Wachtel EF, Keaveny TM. Dependence of trabecular damage on mechanical strain. J Orthop Res. September 1997;15(5):781-787.
2002	Pothuaud L, Van Rietbergen B, Mosekilde L, Beuf O, Levitz P, Benhamou CL, Majumdar S. Combination of topological parameters and bone volume fraction better predicts the mechanical properties of trabecular bone. J Biomech. August 2002;35(8):1091-1099.
2007	Vashishth D. Hierarchy of bone microdamage at multiple length scales. Int J Fatigue. June 2007;29(6):1024-1033.
1995	van Rietbergen B, Weinans H, Huiskes R, Odgaard A. A new method to determine trabecular bone elastic properties and loading using micromechanical finite-element models. J Biomech. January 1995;28(1):69-81.
2001	Kanis JA, Johnell O, Oden A, Dawson A, De Laet C, Jonsson B. Ten year probabilities of osteoporotic fractures according to BMD and diagnostic thresholds. Osteoporos Int. December 2001;12(12):989-995.
2002	Kanis JA. Diagnosis of osteoporosis and assessment of fracture risk. Lancet. June 1, 2002;359(9321):1929-1936.
2000	Vashishth D, Koontz J, Qiu SJ, Lundin-Cannon D, Yeni YN, Schaffler MB, Fyhrie DP. In vivo diffuse damage in human vertebral trabecular bone. Bone. February 2000;26(2):147-152.
1995	Burr DB, Hooser M. Alterations to the en bloc basic fuchsin staining protocol for the demonstration of microdamage produced in vivo. Bone. October 1995;17(4):431-433.
2004	Bushby AJ, Ferguson VL, Boyde A. Nanoindentation of bone: comparison of specimens tested in liquid and embedded in polymethylmethacrylate. J Bone Miner Res. January 2004;19(1):249-259.
1998	Burr DB, Turner CH, Naick P, Forwood MR, Ambrosius W, Hasan MS, Pidaparti R. Does microdamage accumulation affect the mechanical properties of bone? J Biomech. April 1998;31(4):337-345.
1996	Boyde A, Jones SJ. Scanning electron microscopy of bone: instrument, specimen, and issues. Microsc Res Tech. February 1, 1996;33(2):92-120.
1996	Rüegsegger P, Koller B, Müller R. A microtomographic system for the nondestructive evaluation of bone architecture. Calcif Tiss Int. 1996;58(1):24-29.
2007	Perilli E, Baruffaldi F, Visentin M, Bordini B, Traina F, Cappello A, Viceconti M. MicroCT examination of human bone specimens: effects of polymethylmethacrylate embedding on structural parameters. J Microsc (Oxford). 2007;225(pt 2):192-200.
2006	Nazarian A, Stauber M, Zurakowski D, Snyder BD, Müller R. The interaction of microstructure and volume fraction in predicting failure in cancellous bone. Bone. December 2006;39(6):1196-1202.
2007	Nazarian A, Muller J, Zurakowski D, Müller R, Snyder BD. Densitometric, morphometric and mechanical distributions in the human proximal femur. J Biomech. 2007;40(11):2573-2579.
1972	Todd RC, Freeman MAR, Pirie CJ. Isolated trabecular fatigue fractures in the femoral head. J Bone Joint Surg. November 1972;54B(4):723-728.
1985	Burr DB, Martin RB, Schaffler MB, Radin EL. Bone remodeling in response to in vivo fatigue microdamage. J Biomech. 1985;18(3):189-200.
1973	Evans FG. Mechanical Properties of Bone. Springfield, IL: Charles C. Thomas; 1973.
2001	Odgaard A. Quantification of cancellous bone architecture. In: Cowin SC, ed. Bone Mechanics Handbook. 2nd ed. Boca Raton, FL: CRC Press; 2001:chap 14.
2008	Nazarian A, von Stechow D, Zurakowski D, Müller R, Snyder BD. Bone volume fraction explains the variation in strength and stiffness of cancellous bone affected by metastatic cancer and osteoporosis. Calcif Tiss Int. December 2008;83(6):368-379.
1989	Caler WE, Carter DR. Bone creep-fatigue damage accumulation. J Biomech. 1989;22(6-7):625-635.
2004	Loveridge N, Power J, Reeve J, Boyde A. Bone mineralization density and femoral neck fragility. Bone. October 2004;35(4):929-941.
1998	Fazzalari NL, Forwood MR, Manthey BA, Smith K, Kolesik P. Three-dimensional confocal images of microdamage in cancellous bone. Bone. October 1998;23(4):373-378.
1988	Mosekilde L. Age-related changes in vertebral trabecular bone architecture—assessed by a new method. Bone. 1988;9(4):247-250.
1997	Norman TL, Wang Z. Microdamage of human cortical bone: incidence and morphology in long bones. Bone. April 1997;20(4):375-379.
2008	Gupta HS, Zioupos P. Fracture of bone tissue: the ‘hows’ and the ‘whys’. Med Eng Phys. December 2008;30(10):1209-1226.
1974	Whitehouse WJ. The quantitative morphology of anisotropic trabecular bone. J Microsc (Oxford). July 1974;101:153-168.
1977	Frasca P, Harper RA, Katz JL. Collagen fiber orientations in human secondary osteons. Acta Anat. 1977;98(1):1-13.
2004	Hassenkam T, Fantner G, Cutroni J, Weaver J, Morse D, Hansma P. High-resolution AFM imaging of intact and fractured trabecular bone. Bone. July 2004;35(1):4-10.

Year

Entry

1985

Gibson LJ. The mechanical behaviour of cancellous bone. J Biomech. 1985;18(5):317-328.

1973

Vernon-Roberts B, Pirie CJ. Healing trabecular microfractures in the bodies of lumbar vertebrae. Ann Rheum Dis. September 1973;32(5):406-412.

2002

Currey JD. Bones: Structure and Mechanics. Princeton, NJ: Princeton University Press; 2002.

2000

Timlin JA, Carden A, Morris MD, Rajachar RM, Kohn DH. Raman spectroscopic imaging markers for fatigue-related microdamage in bovine bone. Anal Chem. May 15, 2000;72(10):2229-2236.

1983

Brown TD, Shaw DT. In vitro contact stress distributions in the natural human hip. J Biomech. 1983;16(6):373-384.