Osteoporosis is a prevalent bone condition, characterised by low bone mineral density and increased fracture risk. Currently, the gold standard for identifying osteoporosis and increased fracture risk is through quantification of bone mineral density using dual energy X-ray absorption. However, many studies have shown that bone strength, and consequently the probability of fracture, is a combination of both bone mass and bone ‘quality’ (architecture and material chemistry). Although the microarchitecture of both non-fracture and osteoporotic bone has been previously investigated, many of the osteoporotic studies are constrained by factors such as limited sample number, use of ovariectomised animal models, and lack of male and female discrimination. This study reports significant differences in bone quality with respect to the microarchitecture between fractured and non-fractured human femur specimens. Micro-computed tomography was utilised to investigate the microarchitecture of femoral head trabecular bone from a relatively large cohort of non-fracture and fracture human donors. Various microarchitectural parameters have been determined for both groups, providing an understanding of the differences between fracture and non -fracture material. The microarchitecture of non-fracture and fracture bone tissue is shown to be significantly different for many parameters. Differences between sexes also exist, suggesting differences in remodelling between males and females in the fracture group. The results from this study will, in the future, be applied to develop a fracture model which encompasses bone density, architecture and material chemical properties for both female and male tissues.
Table 1 Population characteristics for fracture and non-fracture groups, differentiated according to sex.
Non - Fracture
0.18 ± 0.01
0.18 ± 0.01
0.30 ± 0.01
0.32 ± 0.01
16.06 ± 0.40
17.84 ± 0.53
10.83 ± 0.24
10.10 ± 0.22
0.13 ± 0.004
0.11 ± 0.003
0.19 ± 0.004
0.20 ± 0.005
1.42 ± 0.03
1.60 ± 0.05
1.60 ± 0.02
1.57 ± 0.03
0.60 ± 0.02
0.52 ± 0.02
0.44 ± 0.01
0.45 ± 0.01
1.81 ± 0.06
1.85 ± 0.07
1.14 ± 0.05
1.15 ± 0.06
BMD (g cm-3)
0.30 ± 0.01
0.31 ± 0.02
0.50 ± 0.02
0.52 ± 0.02
TMD (g HA cm-3)
1.61 ± 0.01
1.65 ± 0.01
1.64 ± 0.01
1.62 ± 0.01
Table 2 Average values (in bold) and the associated errors (SEM) for the microarchitectural parameters for fracture and non-fracture groups.
Figure 1. Micro-CT (µ-CT) three-dimensional rendered images from non-fracture (left) and fracture (right) female specimens of the same age (84 yrs).
Figure 2. Relationship between BMD values and age, comparing fracture and non-fracture groups, female specimens (left) and male specimens (right). With age, a significant correlation was observed for both non-fracture males and females. The linear trend correlation coefficients for non-fracture males and females were p < 0.01, R2 = 0.19 and p < 0.05, R2 = 0.14 respectively. Errors have been excluded from the graphs for clarity. For the non-fracture group, each data point represents one donor. For the fracture group each data point represents an individual specimen several of which may arise from a single donor.
Non - Fracture vs Fracture (Age Matched)
Male n = 21 (NF); n = 21 (F)
Female n = 22 (NF); n = 47 (F)
p - value
p - value
0.10 ± 0.02
0.10 ± 0.01
-7.15 ± 0.63
0.07 ± 0.01
0.05 ± 0.01
-0.12 ± 0.06
-0.15 ± 0.05
0.03 ± 0.03
-0.14 ± 0.03
-0.60 ± 0.12
-0.60 ± 0.10
BMD (g cm-3)
0.15 ± 0.03
0.18 ± 0.02
TMD (g HA cm-3)
-0.04 ± 0.02
0.05 ± 0.01
Table 3 P-values for age matched ANOVA analysis of fracture (F) and non-fracture (NF) groups differentiated according to sex, for each microarchitectural parameter.
Linear Regression Analysis
Non - Fracture Correlations with Age
Δ (per 5 yrs)
Δ (per 5 yrs)
-0.007 ± 0.002
-0.007 ± 0.002
0.136 ± 0.065
-0.003 ± 0.001
-0.028 ± 0.006
-0.016 ± 0.006
0.012 ± 0.002
0.009 ± 0.003
BMD (g cm-3)
-0.012 ± 0.004
-0.010 ± 0.004
TMD (g HA cm-3)
Table 4 P-values and R2 calculated from linear regression statistical analysis when comparing the various microarchitecture parameters and age for non-fracture males and females.
Figure 3. Relationship between TbTh values and age, comparing fracture and non-fracture groups, female specimens (left) and male specimens (right). There was no significant correlation with age for non-fracture males, whereas a significant correlation for non-fracture females was observed. The linear trend correlation coefficients for non-fracture females were p < 0.05, R2 = 0.13. Errors have been excluded from the graphs for clarity. For the non-fracture group, each data point represents one donor. For the fracture group each data point represents an individual specimen several of which may arise from a single donor.
Figure 4. Relationship between TbN values and age, comparing fracture and non-fracture groups, female specimens (left) and male specimens (right). With age, a significant correlation was observed for both non-fracture males and females. The linear trend correlation coefficients for non-fracture males and females were p < 0.01, R2 = 0.19 and p < 0.05, R2 = 0.15 respectively. Errors have been excluded from the graphs for clarity. For the non-fracture group, each data point represents one donor. For the fracture group each data point represents an individual specimen several of which may arise from a single donor.
Figure 5. Relationship between TMD values and age, comparing fracture and non-fracture groups, female specimens (left) and male specimens (right). With age, no significant correlation was observed for non-fracture males and females (p > 0.05). Errors have been excluded from the graphs for clarity. For the non-fracture group, each data point represents one donor. For the fracture group each data point represents an individual specimen several of which may arise from a single donor.
Keene GS, Parker MJ, Pryor GA (1993). Mortality and morbidity after hip fractures. BMJ, 307: 1248–1250.
Kanis JA, Johnell O (1999). The burden of osteoporosis. J Endocrinol Invest, 22: 583–588.
Pisani P, Renna MD, Conversano F, Casciaro E, Muratore M, Quarta Eet al. (2013). Screening and early diagnosis of osteoporosis through X-ray and ultrasound-based techniques. World J Radiol, 5: 398–410.
Wilson HC, Abel PD, Shah SIA (2015). Repeated vertebral augmentation for new vertebral compression fractures of postvertebral augmentation patients: a nationwide cohort study-how useful is the current clinical gold standard for fracture risk? Clin Interv Aging, 10: 1653–1655.
Wainwright SA, Marshall LM, Ensrud KE, Cauley JA, Black DM, Hillier TAet al. (2005). Hip fracture in women without osteoporosis. J Clin Endocrinol Metab, 90: 2787–2793.
Faibish D, Ott SM, Boskey AL (2006). Mineral changes in osteoporosis a review. Clin Orthop Relat Res, 443: 28–38.
Teo JC, Si-Hoe KM, Keh JE, Teoh SH (2006). Relationship between CT intensity, microarchitecture and mechanical properties of porcine vertebral cancellous bone. Clin Biomech, 21: 235–244.
Seeman E (2008). Bone quality: the material and structural basis of bone strength. J Bone Mineral Metab, 26: 1–8.
Yerramshetty J, Akkus O (2013) Changes in cortical bone mineral and microstructure with aging and osteoporosis. In: Silva MJ, editor. Skeletal Aging and Osteoporosis. Biomechanics and Mechanobiology. Heidelberg: Springer, 105–131.
Ding M, Hvid I (2000). Quantification of age-related changes in the structure model type and trabecular thickness of human tibial cancellous bone. Bone, 26: 291–295.
Eckstein F, Matsuura M, Kuhn V, Priemel M, Müller R, Link TMet al. (2007). Sex Differences of Human Trabecular Bone Microstructure in Aging Are Site-Dependent. J Bone Miner Res, 22: 817–824.
Thomsen JS, Jensen MV, Niklassen AS, Ebbesen EN, Brüel A (2015). Age-related changes in vertebral and iliac crest 3D bone microstructure-differences and similarities. Osteoporosis Int, 26: 219–228.
Shanbhogue VV, Brixen K, Hansen S (2016). Age-and Sex-Related Changes in Bone Microarchitecture and Estimated Strength: A Three-Year Prospective Study Using HRpQCT. J Bone Miner Res, 31: 1541–1549.
Kijowski R, Tuite M, Kruger D, Munoz Del Rio A, Kleerekoper M, Binkley N (2012). Evaluation of trabecular microarchitecture in non-osteoporotic postmenopausal women with and without fracture. J Bone Miner Res, 27: 1494–1500.
Milovanovic P, Djonic D, Marshall RP, Hahn M, Nikolic S, Zivkovic Vet al. (2012). Micro-structural basis for particular vulnerability of the superolateral neck trabecular bone in the postmenopausal women with hip fractures. Bone, 50: 63–68.
Djuric M, Zagorac S, Milovanovic P, Djonic D, Nikolic S, Hahn Met al. (2013). Enhanced trabecular micro-architecture of the femoral neck in hip osteoarthritis vs. healthy controls: a micro-computer tomography study in postmenopausal women. Int Orthop, 37: 21–26.
Thompson DD, Posner AS, Laughlin WS, Blumenthal NC (1983). Comparison of bone apatite in osteoporotic and normal Eskimos. Calcified Tissue Int, 35: 392–393.
Gadeleta SJ, Boskey AL, Paschalis E, Carlson C, Menschik F, Baldini Tet al. (2000). A physical, chemical, and mechanical study of lumbar vertebrae from normal, ovariectomized, and nandrolone decanoate-treated cynomolgus monkeys (Macaca fascicularis). Bone, 27: 541–550.
Boskey A (2003). Bone mineral crystal size. Osteoporosis Int, 14: 16–21.
Rubin MA, Jasiuk I, Taylor J, Rubin J, Ganey T, Apkarian RP (2003). TEM analysis of the nanostructure of normal and osteoporotic human trabecular bone. Bone, 33: 270–282.
Boskey AL, Donnelly E, Boskey E, Spevak L, Ma Y, Zhang Wet al. (2016). Examining the Relationships Between Bone Tissue Composition, Compositional Heterogeneity, and Fragility Fracture: A Matched Case-Controlled FTIRI Study. J Bone Miner Res, 31: 1070–1081.
Zhang ZM, Li ZC, Jiang LS, Jiang SD, Dai LY (2010). Micro-CT and mechanical evaluation of subchondral trabecular bone structure between postmenopausal women with osteoarthritis and osteoporosis. Osteoporosis Int, 21: 1383–1390.
Li ZC, Dai LY, Jiang LS, Qiu S (2012). Difference in subchondral cancellous bone between postmenopausal women with hip osteoarthritis and osteoporotic fracture: implication for fatigue microdamage, bone microarchitecture, and biomechanical properties. Arthritis Rheum, 64: 3955–3962.
Milovanovic P, Rakocevic Z, Djonic D, Zivkovic V, Hahn M, Nikolic Set al. (2014). Nano-structural, compositional and micro-architectural signs of cortical bone fragility at the superolateral femoral neck in elderly hip fracture patients vs. healthy aged controls. Exp Gerontol, 55: 19–28.
Chiba K, Burghardt AJ, Osaki M, Majumdar S (2013). Heterogeneity of bone microstructure in the femoral head in patients with osteoporosis: an ex vivo HR-pQCT study. Bone, 56: 139–146.
Okazaki N, Chiba K, Taguchi K, Nango N, Kubota S, Ito Met al. (2014). Trabecular microfractures in the femoral head with osteoporosis: Analysis of microcallus formations by synchrotron radiation micro-CT. Bone, 64: 82–87.
Ulrich D, Van Rietbergen B, Laib A, Ruegsegger P (1999). The ability of three-dimensional structural indices to reflect mechanical aspects of trabecular bone. Bone, 25: 55–60.
Bobinac D, Marinovic M, Bazdulj E, Cvijanovic O, Celic T, Maric Iet al. (2013). Microstructural alterations of femoral head articular cartilage and subchondral bone in osteoarthritis and osteoporosis. Osteoarthr Cartilage, 21: 1724–1730.
Manolagas SC (2000). Birth and death of bone cells: basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis. Endocr Rev, 21: 115–137.
Seeman E (1999). The structural basis of bone fragility in men. Bone, 25: 143–147.
Seeman E (2007). Bone’s material and structural strength. Curr Opin Orthop, 18: 494–498.
Gagnon C, Li V, Ebeling PR (2008). Osteoporosis in men: its pathophysiology and the role of teriparatide in its treatment. Clin Interv Aging, 3: 635–45.
Perilli E, Baleani M, Öhman C, Fognani R, Baruffaldi F, Viceconti M (2008). Dependence of mechanical compressive strength on local variations in microarchitecture in cancellous bone of proximal human femur. J Biomech, 41: 438–446.
Sornay-Rendu E, Boutroy S, Duboeuf F, Chapurlat RD (2017). Bone Microarchitecture Assessed by HR-pQCT as Predictor of Fracture Risk in Postmenopausal Women: The OFELY Study. J Bone Miner Res, 32: 1243–1251.
Kreipke TC, Rivera NC, Garrison JG, Easley JT, Turner AS, Niebur GL (2014). Alterations in trabecular bone microarchitecture in the ovine spine and distal femur following ovariectomy. J Biomech, 47: 1918–1921.
Hsu PY, Tsai MT, Wang SP, Chen YJ, Wu J, Hsu JT (2016). Cortical bone morphological and trabecular bone microarchitectural changes in the mandible and femoral neck of ovariectomized rats. PloS One, 11: e154367.
Liu H, Li W, Liu YS, Zhou YS (2016). Bone micro-architectural analysis of mandible and tibia in ovariectomised rats. Bone Joint Res, 5: 253–62.
Vale AC, Pereira MF, Maurício A, Amaral P, Rosa LG, Lopes Aet al. (2013). Micro-computed tomography and compressive characterization of trabecular bone. Colloids Surf A Physicochem Eng Asp, 438: 199–205.
Greenwood C, Clement J, Dicken A, Evans JP, Lyburn I, Martin RMet al. (2016). Towards new material biomarkers for fracture risk. Bone, 93:55–63.
Doube M, Kłosowski MM, Arganda-Carreras I, Cordeliéres F, Dougherty RP, Jackson Jet al. (2010). BoneJ: free and extensible bone image analysis in ImageJ. Bone, 47:1076–9.
Salmon PL, Ohlsson C, Shefelbine SJ, Doube M. (2015). Structure model index does not measure rods and plates in trabecular bone. Front Endocrinol, 6:1–10.
Nazarian A, Muller J, Zurakowski D, Müller R, Snyder BD (2007). Densitometric, morphometric and mechanical distributions in the human proximal femur. J Biomech, 40: 2573–2579.
Müller, R, Rüegsegger P (1997). Micro-tomographic imaging of non-destructive evaluation of trabecular bone architecture In: Lowet G, Rüegsegger P, Weinans H, Meunier A, editors. Bone Research in Biomechanics. Amsterdam: IOS Press, 61–80.
Hildebrand T, Laib A, Müller R, Dequeker J, Rüegsegger P (1999). Direct three-dimensional morphometric analysis of human cancellous bone: Microstructural data from spine, femur, iliac crest, and calcaneus. J Bone Miner Res, 14: 1167–1174.
Öhman C, Baleani M, Perilli E, Dall’Ara E, Tassani S, Baruffaldi, F et al. (2007). Mechanical testing of cancellous bone from the femoral head: experimental errors due to off-axis measurements. J Biomech, 40: 2426–2433.
Tassani S, Particelli F, Perilli E, Traina F, Baruffaldi F, Viceconti M (2011). Dependence of trabecular structure on bone quantity: a comparison between osteoarthritic and non-pathological bone. Clin Biomech, 26: 632–639.
Nikodem, A (2012). Correlations between structural and mechanical properties of human trabecular femur bone. Acta Bioeng Biomech, 14: 37–47.
Macdonald HM, Nishiyama KK, Kang J, Hanley DA, Boyd SK (2011). Age-related patterns of trabecular and cortical bone loss differ between sexes and skeletal sites: A population-based HR-pQCT study. J Bone Miner Res, 26: 50–62.
Humbert L, Whitmarsh T, Craene MD, Del Río Barquero LM, Frangi AF (2012). Technical Note: Comparison between single and multiview simulated DXA configurations for reconstructing the 3D shape and bone mineral density distribution of the proximal femur. Med Phys, 39: 5272–5276.
Warming L, Hassager C, Christiansen C (2002). Changes in bone mineral density with age in men and women: A longitudinal study. Osteoporosis Int, 13: 105–112.
Greenwood C, Clement JG, Dicken AJ, Evans JPO, Lyburn ID, Martin RMet al. (2015). The micro-architecture of human cancellous bone from fracture neck of femur patients in relation to the structural integrity and fracture toughness of the tissue. Bone Reports, 3: 67–75.
Akkus O, Adar F, Schaffler MB (2004). Age-related changes in physicochemical properties of mineral crystals are related to impaired mechanical function of cortical bone. Bone, 34: 443–453.
Handschin RG, Stern WB (1995). X-ray diffraction studies on the lattice perfection of human bone apatite (Crista iliaca). Bone, 16: S355–S363
Acerbo AS, Kwaczala AT, Yang L, Judex S, Miller LM (2014). Alterations in collagen and mineral nanostructure observed in osteoporosis and pharmaceutical treatments using simultaneous small-and wide-angle X-ray scattering. Calcified Tissue Int, 95: 446–456.
Rey C, Renugopalakrishman V, Collins B, Glimcher MJ (1991). Fourier transform infrared spectroscopic study of the carbonate ions in bone mineral during aging. Calcified Tissue Int, 49: 251–258.
Posner AS, Perloff A, Diorio AF (1958). Refinement of the hydroxyapatite structure. Acta Crystallogr, 11: 308–309.
El Feki H, Savariault JM, Salah AB, Jemal M. (2000). Sodium and carbonate distribution in substituted calcium hydroxyapatite. Solid State Sci, 2: 577–586.
Wilson RM, Elliott JC, Dowker SE, Smith RI. (2004). Rietveld structure refinement of precipitated carbonate apatite using neutron diffraction data. Biomaterials, 25: 2205–2213.
Paschalis EP, Betts F, DiCarlo E, Mendelsohn R, Boskey AL (1997). FTIR microspectroscopic analysis of human iliac crest biopsies from untreated osteoporotic bone. Calcified Tissue Int, 61: 487–492.
Huang RY, Miller LM, Carlson CS, Chance MR (2003). In situ chemistry of osteoporosis revealed by synchrotron infrared microspectroscopy. Bone, 33: 514–521.