Registered Micro-Computed Tomography Data as a Four-Dimensional Imaging Biomarker of Bone Formation and Resorption

  • Annette I. Birkhold
  • Bettina M. WillieEmail author
Reference work entry
Part of the Biomarkers in Disease: Methods, Discoveries and Applications book series (BDMDA)


There are significant clinical reasons motivating scientists to better understand how loading conditions, diseases, synthetic implants, and drug treatments affect bone formation and resorption. Changes in bone turnover have enormous impact on the quality and mechanical competence of the skeleton. Until recently, bone formation and resorption were primarily measured using biochemical markers of bone turnover or histomorphometry. However, recent advances in computed tomography allow one to follow structural changes in the cortical and trabecular bone of living animals and human patients. The aim of this chapter is to describe recently developed methods that allow the monitoring of bone modeling and remodeling processes in vivo by using registered longitudinal micro-computed tomography data, which serves as an imaging biomarker of bone formation and resorption. The chapter provides an overview of bone modeling and remodeling processes and the standard methods that have been used in the past and present to assess bone formation and resorption. Micro-computed tomography-based imaging of the bone is then discussed. A detailed description is then given of recently developed computation methods that allow monitoring of bone modeling and remodeling using registered longitudinal micro-computed tomography data as an imaging biomarker of bone formation and resorption. The chapter ends with a discussion of how these imaging-based biomarkers of formation and resorption can be used to complement and in some cases replace conventional experimental and clinical methods of monitoring bone turnover.


Bone formation Bone resorption Remodeling Modeling Bone turnover Imaging biomarker Micro-computed tomography 

List of Abbreviations


Activation, formation, and resorption


Bone-specific alkaline phosphatase


Bone formation rate


Basic multicellular unit


Bone resorption rate


Bone surface


Bone sialoprotein


Bone volume


Carboxy-terminal cross-linked telopeptide of type I collagen




Eroded surface


Eroded volume


High-resolution peripheral quantitative computed tomography


Lipoprotein receptor-related protein 5 and 6


Mineral apposition rate


Micro-computed tomography


Mineral resorption rate


Mineralizing surface


Mineralized volume


Amino-terminal cross-linked telopeptide of type I collagen




Procollagen type I C-terminal propeptide


Procollagen type I N-terminal propeptide




5b isoenzyme of tartrate-resistant acid phosphatase


Tartrate-resistant acid phosphatase


  1. Birkhold AI, Razi H, Duda GN, et al. The influence of age on adaptive bone formation and bone resorption. Biomaterials. 2014a;35:9290–301.CrossRefPubMedGoogle Scholar
  2. Birkhold AI, Razi H, Duda GN, et al. Mineralizing surface is the main target of mechanical stimulation independent of age: 3D dynamic in vivo morphometry. Bone. 2014b;66:15–25.CrossRefPubMedGoogle Scholar
  3. Birkhold AI, Razi H, Weinkamer R, et al. Monitoring in vivo (re)modeling: a computational approach using 4D microCT data to quantify bone surface movements. Bone. 2015;75:210–21.CrossRefPubMedGoogle Scholar
  4. Boone JM, Velazquez O, Cherry SR. Small-animal X-ray dose from micro-CT. Mol Imaging. 2004;3:149–58.CrossRefPubMedGoogle Scholar
  5. Bouxsein ML, Boyd SK, Christiansen BA, et al. Guidelines for assessment of bone microstructure in rodents using micro-computed tomography. J Bone Miner Res. 2010;25:1468–86.CrossRefPubMedGoogle Scholar
  6. Boyd SK, Davison P, Mueller R, et al. Monitoring individual morphological changes over time in ovariectomized rats by in vivo micro-computed tomography. Bone. 2006;39:854–62.CrossRefPubMedGoogle Scholar
  7. Buie HR, Campbell GM, Klinck RJ, et al. Automatic segmentation of cortical and trabecular compartments based on a dual threshold technique for in vivo micro-CT bone analysis. Bone. 2007;41:505–15.CrossRefPubMedGoogle Scholar
  8. Buie HR, Moore CP, Boyd SK. Postpubertal architectural developmental patterns differ between the L3 vertebra and proximal tibia in three inbred strains of mice. J Bone Miner Res. 2008;23:2048–59.CrossRefPubMedGoogle Scholar
  9. Burghardt AJ, Kazakia GJ, Majumdar S. A local adaptive threshold strategy for high resolution peripheral quantitative computed tomography of trabecular bone. Ann Biomed Eng. 2007;35:1678–86.CrossRefPubMedGoogle Scholar
  10. Carlson SK, Classic KL, Bender CE, et al. Small animal absorbed radiation dose from serial micro-computed tomography imaging. Mol Imaging Biol. 2007;9:78–82.CrossRefPubMedGoogle Scholar
  11. Chappard C, Basillais A, Benhamou L, et al. Comparison of synchrotron radiation and conventional x-ray microCT for assessing trabecular bone microarchitecture of human femoral heads. Med Phys. 2006;33:3568–77.CrossRefPubMedGoogle Scholar
  12. Christen P, Ito K, Ellouz R, et al. Bone remodelling in humans is load-driven but not lazy. Nat Commun. 2014;5:4855.CrossRefPubMedGoogle Scholar
  13. Cormack AM. Representation of a function by its line integrals, with some radiological applications. New York: American Institute of Physics; 1963.Google Scholar
  14. David V, Laroche N, Boudignon B, et al. Noninvasive in vivo monitoring of bone architecture alterations in hindlimb-unloaded female rats using novel three-dimensional microcomputed tomography. J Bone Miner Res. 2003;18:1622–31.CrossRefPubMedGoogle Scholar
  15. Dempster DW, Compston JE, Drezner MK, et al. Standardized nomenclature, symbols, and units for bone histomorphometry. J Bone Miner Res. 2013;28:2–17.CrossRefPubMedPubMedCentralGoogle Scholar
  16. Deserno TM. Biomedical image processing. Berlin/Heidelberg: Springer; 2011.CrossRefGoogle Scholar
  17. Dufresne T. Segmentation techniques for analysis of bone by three-dimensional computed tomographic imaging. Technol Health Care. 1998;6:351–9.PubMedGoogle Scholar
  18. Duyar I, Pelin C. Body height estimation based on tibia length in different stature groups. Am J Phys Anthropol. 2003;122:23–7.CrossRefPubMedGoogle Scholar
  19. Elliott JC, Dover SD. X-ray microtomography. J Microsc. 1982;126:211–3.CrossRefPubMedGoogle Scholar
  20. Erben RG. Trabecular and endocortical bone surfaces in the rat: modeling or remodeling? Anat Rec. 1996;246:39–46.CrossRefPubMedGoogle Scholar
  21. Erben RG, Glösmann M. Chapter 19: Histomorphometry in rodents. In: Bone research protocols. Methods in molecular biology. 2nd ed. New York: Springer; 2012.Google Scholar
  22. Eriksen EF. Normal and pathological remodeling of human trabecular bone: three dimensional reconstruction of the remodeling sequence in normals and in metabolic bone disease. Endocr Rev. 1986;7:379–408.CrossRefPubMedGoogle Scholar
  23. Eriksen EF, Gundersen HJ, Melsen F, et al. Reconstruction of the formative site in iliac trabecular bone in 20 normal individuals employing a kinetic model for matrix and mineral apposition. Metab Bone Dis Relat Res. 1984a;5:243–52.CrossRefPubMedGoogle Scholar
  24. Eriksen EF, Melsen F, Mosekilde L. Reconstruction of the resorptive site in iliac trabecular bone: a kinetic model for bone resorption in 20 normal individuals. Metab Bone Dis Relat Res. 1984b;5:235–42.CrossRefPubMedGoogle Scholar
  25. Feldkamp LA, Goldstein SA, Parfitt AM, et al. The direct examination of 3D bone architecture in vitro by CT. J Bone Miner Res. 1989;4:3–11.CrossRefPubMedGoogle Scholar
  26. Ford NL, Thornton MM, Holdsworth DW. Fundamental image quality limits for microcomputed tomography in small animals. Med Phys. 2003;30:2869–77.CrossRefPubMedGoogle Scholar
  27. Franz-Odendaal TA, Hall BK, Witten PE. Buried alive: how osteoblasts become osteocytes. Dev Dyn. 2006;235:176–90.CrossRefPubMedGoogle Scholar
  28. Frost HM. Tetracycline-based histological analysis of bone remodeling. Calcif Tissue Res. 1969;3:211–37.CrossRefPubMedGoogle Scholar
  29. Gelaude F, Vander Sloten J, Lauwers B. Semi-automated segmentation and visualisation of outer bone cortex from medical images. Comput Methods Biomech Biomed Engin. 2006;9:65–77.CrossRefPubMedGoogle Scholar
  30. Gonzalez EA, Lund RJ, Martin KJ, et al. Treatment of a murine model of high-turnover renal osteodystrophy by exogenous BMP-7. Kidney Int. 2002;61:1322–31.CrossRefPubMedGoogle Scholar
  31. Halloran BP, Ferguson VL, Simske SJ, et al. Changes in bone structure and mass with advancing age in the male C57BL/6J mouse. J Bone Miner Res. 2002;17:1044–50.CrossRefPubMedGoogle Scholar
  32. Hattner R, Epker BN, Frost HM. Suggested sequential mode of control of changes in cell behaviour in adult bone remodelling. Nature. 1965;206:489–90.CrossRefPubMedGoogle Scholar
  33. Herman GT. Fundamentals of computerized tomography: image reconstruction from projections. Dordrecht/New York: Springer; 2009.CrossRefGoogle Scholar
  34. Hernandez CJ, Hazelwood SJ, Martin RB. The relationship between basic multicellular unit activation and origination in cancellous bone. Bone. 1999;25:585–7.CrossRefPubMedGoogle Scholar
  35. Hlaing TT, Compston JE. Biochemical markers of bone turnover – uses and limitations. Ann Clin Biochem. 2014;51:189–202.CrossRefPubMedGoogle Scholar
  36. Hounsfield GN. Computerized transverse axial scanning (tomography). Br J Radiol. 1973;46:1016–22.CrossRefPubMedGoogle Scholar
  37. Jaworski ZF, Lok E. The rate of osteoclastic bone erosion in Haversian remodeling sites of adult dog’s rib. Calcif Tissue Res. 1972;10:103–12.CrossRefPubMedGoogle Scholar
  38. Kettenberger U, Ston J, Thein E, et al. Does locally delivered Zoledronate influence peri-implant bone formation? - Spatio-temporal monitoring of bone remodeling in vivo. Biomaterials. 2014;35:9995–10006.CrossRefPubMedGoogle Scholar
  39. Klinck RJ, Campbell GM, Boyd SK. Radiation effects on bone architecture in mice and rats resulting from in vivo microCT scanning. Med Eng Phys. 2008;30:888–95.CrossRefPubMedGoogle Scholar
  40. Kohler T, Stauber M, Donahue LR, et al. Automated compartmental analysis for high-throughput skeletal phenotyping in femora of genetic mouse models. Bone. 2007;41:659–67.CrossRefPubMedGoogle Scholar
  41. Lukas C, Ruffoni D, Lambers FM, et al. Mineralization kinetics in murine trabecular bone quantified by time-lapsed in vivo microCT. Bone. 2013;56:55–60.CrossRefPubMedGoogle Scholar
  42. Manolagas SC. Birth and death of bone cells: basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis. Endocr Rev. 2000;21:115–37.PubMedGoogle Scholar
  43. Martin RB, Burr DB, Sharkey NA. Skeletal tissue mechanics. New York: Springer; 1998.CrossRefGoogle Scholar
  44. Meganck JA, Kozloff KM, Thornton MM, et al. Beam hardening artifacts in microCT scanning can be reduced by X-ray beam filtration and the resulting images can be used to accurately measure BMD. Bone. 2009;45:1104–16.CrossRefPubMedPubMedCentralGoogle Scholar
  45. Meijering E. Spline interpolation in medical imaging: comparison with other convolution-based approaches. Signal Processing X: Theories and Applications Proceedings of EUSIPCO; 2000. 2000:1989–1996Google Scholar
  46. Milch RA, Rall DP, Tobie JE. Fluorescence of tetracycline antibiotics in bone. J Bone Joint Surg Am. 1958;40-A:897–910.CrossRefPubMedGoogle Scholar
  47. Nishiyama KK, Campbell GM, Klinck RJ, et al. Reproducibility of bone micro-architecture measurements in rodents by in vivo microCT is maximized with 3D image registration. Bone. 2010;66:155–61.CrossRefGoogle Scholar
  48. Nuzzo S, Lafage-Proust MH, Martin-Badosa E, et al. Synchrotron radiation microtomography allows the analysis of three-dimensional microarchitecture and degree of mineralization of human iliac crest biopsy specimens: effects of etidronate treatment. J Bone Miner Res. 2002a;17:1372–82.CrossRefPubMedGoogle Scholar
  49. Nuzzo S, Peyrin F, Cloetens P, et al. Quantification of the degree of mineralization of bone in three dimensions using synchrotron radiation microtomography. Med Phys. 2002b;29:2672–81.CrossRefPubMedGoogle Scholar
  50. Oliveira F, Tavares J. Medical image registration: a review. Comput Methods Biomech Biomed Engin. 2014;17:73–93.CrossRefPubMedGoogle Scholar
  51. Parfitt AM. Age-related structural changes in trabecular and cortical bone: cellular mechanisms and biomechanical consequences. Calcif Tissue Int. 1984;36 Suppl 1:S123–8.CrossRefPubMedGoogle Scholar
  52. Parfitt AM. Osteonal and hemi-osteonal remodeling: the spatial and temporal framework for signal traffic in adult human bone. J Cell Biochem. 1994;55:273–86.CrossRefPubMedGoogle Scholar
  53. Parfitt AM. Osteoclast precursors as leukocytes: importance of the area code. Bone. 1998;23:491–4.CrossRefPubMedGoogle Scholar
  54. Parfitt AM, Mathews CH, Villanueva AR, et al. 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. 1983;72:1396–409.CrossRefPubMedPubMedCentralGoogle Scholar
  55. Parfitt AM, Drezner MK, Glorieux FH, et al. Bone histomorphometry: standardization of nomenclature, symbols, and units. J Bone Miner Res. 1987;2:595–610.CrossRefPubMedGoogle Scholar
  56. Pluim JPW, Maintz JBA, Viergever MA. Mutual-information-based registration of medical images: a survey. IEEE Trans Med Imaging. 2003;22:986–1004.CrossRefPubMedGoogle Scholar
  57. Radon J. Über die Bestimmung von Funktionen durch ihre Integralwerte längs gewisser Mannigfaltigkeiten. Akad Wiss. 1971;69:262–77.Google Scholar
  58. Razi H, Birkhold AI, Weinkamer R, et al. Aging leads to a dysregulation in mechanically driven bone formation and resorption. J Bone Miner Res. 2015;30:1864–1873.CrossRefPubMedGoogle Scholar
  59. Recker RR. Bone histomorphometry: techniques and interpretation. Boca Raton: CRC Press; 1983.Google Scholar
  60. Riggs BL, Melton LJ, Robb RA, et al. A population-based assessment of rates of bone loss at multiple skeletal sites: evidence for substantial trabecular bone loss in young adult women and men. J Bone Miner Res. 2008;23:205–14.CrossRefPubMedGoogle Scholar
  61. Rueegsegger P, Koller B, Mueller R. A microtomographic system for the nondestructive evaluation of bone architecture. Calcif Tissue Int. 1996;58:24–9.CrossRefGoogle Scholar
  62. Schulte FA, Lambers FM, Kuhn G, et al. In vivo microCT allows direct 3D quantification of both bone formation and bone resorption parameters using time-lapsed imaging. Bone. 2011;48:433–42.CrossRefPubMedGoogle Scholar
  63. Schulte FA, Ruffoni D, Lambers FM, et al. Local mechanical stimuli regulate bone formation and resorption in mice at the tissue level. PLoS One. 2013;8:e62172.CrossRefPubMedPubMedCentralGoogle Scholar
  64. Slyfield CR, Tkachenko EV, Wilson DL, et al. Three-dimensional dynamic bone histomorphometry. J Bone Miner Res. 2012;27:486–95.CrossRefPubMedPubMedCentralGoogle Scholar
  65. Somerville JM, Aspden RM, Armour KE, et al. Growth of C57BL/6 mice and the material and mechanical properties of cortical bone from the tibia. Calcif Tissue Int. 2004;74:469–75.CrossRefPubMedGoogle Scholar
  66. Thévenaz P, Ruttimann UE, Unser M. A pyramid approach to subpixel registration based on intensity. IEEE Trans Image Process. 1998;7:27–41.CrossRefPubMedGoogle Scholar
  67. Vasikaran S, Eastell R, Bruyere O, et al. Markers of bone turnover for the prediction of fracture risk and monitoring of osteoporosis treatment: a need for international reference standards. Osteoporos Int. 2011;22:391–420.CrossRefPubMedGoogle Scholar
  68. Waarsing JH, Day JS, van der Linden JC, et al. Detecting and tracking local changes in the tibiae of individual rats. Bone. 2004a;34:163–9.CrossRefPubMedGoogle Scholar
  69. Waarsing JH, Day JS, Weinans H. An improved segmentation method for in vivo microCT imaging. J Bone Miner Res. 2004b;19:1640–50.CrossRefPubMedGoogle Scholar
  70. Weinstein RS, Jilka RL, Parfitt A, et al. Inhibition of osteoblastogenesis and promotion of apoptosis of osteoblasts and osteocytes by glucocorticoids. J Clin Investig. 1998;102:274–82.CrossRefPubMedPubMedCentralGoogle Scholar
  71. Weinstein RS, Chen JR, Powers CC, et al. Promotion of osteoclast survival and antagonism of bisphosphonate-induced osteoclast apoptosis by glucocorticoids. J Clin Investig. 2002;109:1041–8.CrossRefPubMedPubMedCentralGoogle Scholar
  72. Wells WM, Viola P, Atsumi H, et al. Multi-modal volume registration by maximization of mutual information. Med Image Anal. 1996;1:35–51.CrossRefPubMedGoogle Scholar
  73. Willie BM, Birkhold AI, Razi H, et al. Diminished response to in vivo mechanical loading in trabecular and not cortical bone in adulthood of female C57Bl/6 mice coincides with a reduction in deformation to load. Bone. 2013;55:335–46.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Continuum Biomechanics and Mechanobiology Research GroupInstitute of Applied Mechanics, University of StuttgartStuttgartGermany
  2. 2.Department of Pediatric Surgery, McGill UniversityResearch Centre, Shriners Hospital for Children-CanadaMontrealCanada

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