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. 2000 Oct;197(Pt 3):413–420. doi: 10.1046/j.1469-7580.2000.19730413.x

Visualisation of three-dimensional microcracks in compact bone

F J O'BRIEN 1 ,2 , D TAYLOR 2 , G R DICKSON 3 , T C LEE 1 ,2 ,✉
PMCID: PMC1468142  PMID: 11117627

Abstract

Microdamage in bone contributes to the loss of bone quality in osteoporosis and is thought to play a major role in both fragility and stress fractures (Schaffler et al. 1995). In this study, in vivo microcracks in human ribs were bulk-stained in basic fuchsin and viewed in longitudinal section and in 3 dimensions using 2 different computer-based methods of reconstruction: (1) serial sectioning of methylmethacrylate embedded sections using a sledge macrotome and identification of microcracks using UV epifluorescence followed by computerised reconstruction of microcracks using software and (2) laser scanning confocal microscopy of thick sections followed by reconstruction of microcracks into a 3-D image. The size and shape of microcracks were found to be similar using both techniques. Both techniques of reconstruction showed microcracks to be approximately elliptical in shape. From the serial sectioning reconstructions (n = 9), microcracks were found to have a mean length of 404±145 μm (mean± S.D.) (in the longitudinal direction) and mean width of 97±38 μm (in the transverse direction). Using epifluorescence microscopy, 92 microcracks were identified; mean microcrack length was 349±100 μm in the longitudinal direction. This was consistent with other results (Burr & Martin, 1993) and with the theoretical prediction of an elliptical crack shape with aspect ratio (longitudinal∶transverse) of 5∶1 deduced from analysis of random 2-D sections (Taylor & Lee, 1998). The results obtained provide new data on the nature of microcracks in bone and the method has the potential to become a useful tool in the calculation of stress intensity values which indicate the probability of an individual microcrack propagating to cause a stress or fragility fracture.

Keywords: Epifluorescence microscopy, laser scanning confocal microscopy, osteoporosis, stress fractures, fragility fractures

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Selected References

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  1. Bentolila V., Boyce T. M., Fyhrie D. P., Drumb R., Skerry T. M., Schaffler M. B. Intracortical remodeling in adult rat long bones after fatigue loading. Bone. 1998 Sep;23(3):275–281. doi: 10.1016/s8756-3282(98)00104-5. [DOI] [PubMed] [Google Scholar]
  2. Boyce T. M., Fyhrie D. P., Glotkowski M. C., Radin E. L., Schaffler M. B. Damage type and strain mode associations in human compact bone bending fatigue. J Orthop Res. 1998 May;16(3):322–329. doi: 10.1002/jor.1100160308. [DOI] [PubMed] [Google Scholar]
  3. Burr D. B., Hooser M. Alterations to the en bloc basic fuchsin staining protocol for the demonstration of microdamage produced in vivo. Bone. 1995 Oct;17(4):431–433. doi: 10.1016/s8756-3282(95)00241-3. [DOI] [PubMed] [Google Scholar]
  4. Burr D. B., Martin R. B. Calculating the probability that microcracks initiate resorption spaces. J Biomech. 1993 Apr-May;26(4-5):613–616. doi: 10.1016/0021-9290(93)90023-8. [DOI] [PubMed] [Google Scholar]
  5. Burr D. B., Martin R. B., Schaffler M. B., Radin E. L. Bone remodeling in response to in vivo fatigue microdamage. J Biomech. 1985;18(3):189–200. doi: 10.1016/0021-9290(85)90204-0. [DOI] [PubMed] [Google Scholar]
  6. Fazzalari N. L., Forwood M. R., Manthey B. A., Smith K., Kolesik P. Three-dimensional confocal images of microdamage in cancellous bone. Bone. 1998 Oct;23(4):373–378. doi: 10.1016/s8756-3282(98)00111-2. [DOI] [PubMed] [Google Scholar]
  7. Fyhrie D. P., Schaffler M. B. Failure mechanisms in human vertebral cancellous bone. Bone. 1994 Jan-Feb;15(1):105–109. doi: 10.1016/8756-3282(94)90900-8. [DOI] [PubMed] [Google Scholar]
  8. Lee T. C., Arthur T. L., Gibson L. J., Hayes W. C. Sequential labelling of microdamage in bone using chelating agents. J Orthop Res. 2000 Mar;18(2):322–325. doi: 10.1002/jor.1100180222. [DOI] [PubMed] [Google Scholar]
  9. Lee T. C., Myers E. R., Hayes W. C. Fluorescence-aided detection of microdamage in compact bone. J Anat. 1998 Aug;193(Pt 2):179–184. doi: 10.1046/j.1469-7580.1998.19320179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lee T. C., Taylor D. Bone remodelling: should we cry Wolff? Ir J Med Sci. 1999 Apr-Jun;168(2):102–105. doi: 10.1007/BF02946474. [DOI] [PubMed] [Google Scholar]
  11. Martin R. B., Burr D. B. A hypothetical mechanism for the stimulation of osteonal remodelling by fatigue damage. J Biomech. 1982;15(3):137–139. doi: 10.1016/s0021-9290(82)80001-8. [DOI] [PubMed] [Google Scholar]
  12. Martin R. B. Toward a unifying theory of bone remodeling. Bone. 2000 Jan;26(1):1–6. doi: 10.1016/s8756-3282(99)00241-0. [DOI] [PubMed] [Google Scholar]
  13. McBryde A. M., Jr Stress fractures in athletes. J Sports Med. 1975 Sep-Oct;3(5):212–217. doi: 10.1177/036354657500300502. [DOI] [PubMed] [Google Scholar]
  14. Prendergast P. J., Taylor D. Prediction of bone adaptation using damage accumulation. J Biomech. 1994 Aug;27(8):1067–1076. doi: 10.1016/0021-9290(94)90223-2. [DOI] [PubMed] [Google Scholar]
  15. Riggs B. L., Melton L. J., 3rd The worldwide problem of osteoporosis: insights afforded by epidemiology. Bone. 1995 Nov;17(5 Suppl):505S–511S. doi: 10.1016/8756-3282(95)00258-4. [DOI] [PubMed] [Google Scholar]
  16. Schaffler M. B., Choi K., Milgrom C. Aging and matrix microdamage accumulation in human compact bone. Bone. 1995 Dec;17(6):521–525. doi: 10.1016/8756-3282(95)00370-3. [DOI] [PubMed] [Google Scholar]
  17. Schaffler M. B., Radin E. L., Burr D. B. Mechanical and morphological effects of strain rate on fatigue of compact bone. Bone. 1989;10(3):207–214. doi: 10.1016/8756-3282(89)90055-0. [DOI] [PubMed] [Google Scholar]
  18. Sherman S., Hadley E. C. Aging and bone quality: an underexplored frontier. Calcif Tissue Int. 1993;53 (Suppl 1):S1–S1. doi: 10.1007/BF01673392. [DOI] [PubMed] [Google Scholar]
  19. Taylor D., Lee T. C. Measuring the shape and size of microcracks in bone. J Biomech. 1998 Dec;31(12):1177–1180. doi: 10.1016/s0021-9290(98)00133-x. [DOI] [PubMed] [Google Scholar]
  20. Taylor D. Microcrack growth parameters for compact bone deduced from stiffness variations. J Biomech. 1998 Jul;31(7):587–592. doi: 10.1016/s0021-9290(98)00050-5. [DOI] [PubMed] [Google Scholar]

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