Skip to main content
SpringerLink
Log in

Anisotropic aspects of solubility behavior in the demineralization of cortical bone revealed by XRD analysis

  • Original Paper
  • Published:
Journal of Biological Physics Aims and scope Submit manuscript

Abstract

Dissolution of cortical bone mineral under demineralization in 0.1 M HCl and 0.1 M EDTA solutions is studied by X-ray diffraction (XRD). The bone specimens (in the form of planar oriented pieces) were cut from a diaphysial fragment of a mature mammal bone so that a cross-section surface and a longitudinal section surface could be analyzed individually. This permitted to compare the dissolution behavior of bone apatite of different morphologies: crystals having the c-axis of the hexagonal unit-cell generally parallel to the long axis of the bone (major morphology) and those having the c-axis almost perpendicular to the bone axis (minor morphology). For these two types of morphology, the crystallite sizes in two mutually perpendicular directions (namely, [002] and [310]) were estimated by Scherrer formula in the initial and the stepwise-demineralized specimens. The data obtained reveal that the crystals belonging to the minor morphology dissolve faster than the crystals of the major morphological type, despite the fact that the crystallites of the minor morphology seem to be only a little smaller than those of the major morphology; the apatite crystallites irrespective of the morphology type are elongated in the c-axis direction. We hypothesize that the revealed difference in solubility may be caused by diverse chemical modifications of apatite of these two morphological types, since the solubility of apatite is strictly regulated by anionic and cationic substitutions in the lattice. The anisotropy effect in solubility of bone mineral seems to be functionally predetermined and this should be a crucial factor in the resorption and remodeling behavior of a bone. Some challenges arising at XRD examination of partially decalcified cortical bone blocks are discussed, as well as the limitations of estimation of bone crystallite size by XRD line-broadening analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. LeGeros, R.Z.: Apatites in biological systems. Prog. Cryst. Growth Charact. Mater. 4(1), 1–45 (1981)

    Google Scholar 

  2. Rey, C., Combes, C., Drouet, C., Glimcher, M.J.: Bone mineral: update on chemical composition and structure. Osteoporos. Int. 20(6), 1013–1021 (2009)

    Article  Google Scholar 

  3. Combes, C., Cazalbou, S., Rey, C.: Apatite biominerals. Minerals 6(2), 34 (2016). https://doi.org/10.3390/min6020034

    Article  Google Scholar 

  4. Robinson, R.A.: An electron-microscopic study of the crystalline inorganic component of bone and its relationship to the organic matrix. J. Bone Joint Surg. Am. 34(2), 389–476 (1952)

    Article  Google Scholar 

  5. Eppell, S.J., Tong, W., Katz, J.L., Kuhn, L., Glimcher, M.J.: Shape and size of isolated bone mineralites measured using atomic force microscopy. J. Orthop. Res. 19(6), 1027–1034 (2001)

    Article  Google Scholar 

  6. Carlström, D., Glas, J.E.: The size and shape of the apatite crystallites in bone as determined from line-broadening measurements on oriented specimens. Biochim. Biophys. Acta 35, 46–53 (1959)

    Article  Google Scholar 

  7. Handschin, R.G., Stern, W.B.: X-ray diffraction studies on the lattice perfection of human bone apatite (Crista iliaca). Bone 16(4), S355–S363 (1995)

    Article  Google Scholar 

  8. Rindby, A., Voglis, P., Engström, P.: Microdiffraction studies of bone tissues using synchrotron radiation. Biomaterials 19(22), 2083–2090 (1998)

    Article  Google Scholar 

  9. Sakae, T., Kono, T., Okada, H., Nakada, H., Ogawa, H., Tsukioka, T., Kaneda, T.: X-ray micro-diffraction analysis revealed the crystallite size variation in the neighboring regions of a small bone mass. J. Hard Tissue Biol. 26(1), 103–107 (2017)

    Article  Google Scholar 

  10. Lees, S.: A model for the distribution of HAP crystallites in bone—an hypothesis. Calci. Tissue Int. 27(1), 53–56 (1979)

    Article  Google Scholar 

  11. Matsushima, N., Akiyama, M., Terayama, Y.: Quantitative analysis of the orientation of mineral in bone from small-angle X-ray scattering patterns. Jpn. J. Appl. Phys. 21(1), 186–189 (1982)

    Article  ADS  Google Scholar 

  12. Fratzl, P., Schreiber, S., Klaushofer, K.: Bone mineralization as studied by small-angle X-ray scattering. Connect. Tissue Res. 34(4), 247–254 (1996)

    Article  Google Scholar 

  13. Wenk, H.R., Heidelbach, F.: Crystal alignment of carbonated apatite in bone and calcified tendon: results from quantitative texture analysis. Bone 24(4), 361–369 (1999)

    Article  Google Scholar 

  14. Sasaki, N., Matsushima, N., Ikawa, T., Yamamura, H., Fukuda, A.: Orientation of bone mineral and its role in the anisotropic mechanical properties of bone: transverse anisotropy. J. Biomech. 22(2), 157–164 (1989)

    Article  Google Scholar 

  15. Sasaki, N., Sudoh, Y.: X-ray pole figure analysis of apatite crystals and collagen molecules in bone. Calcif. Tissue Int. 60(4), 361–367 (1997)

    Article  Google Scholar 

  16. Klug, H., Alexander, L.: X-Ray Diffraction Procedure for Polycrystallite and Amorphous Materials. Wiley, New York (1974)

    Google Scholar 

  17. Boskey, A.: Bone mineral crystal size. Osteoporos. Int. 14, S16–S21 (2003)

    Article  Google Scholar 

  18. Horvath, A.L.: Solubility of structurally complicated materials: II. Bone. J. Phys. Chem. Ref. Data 35(4), 1653–1668 (2006)

    Article  ADS  Google Scholar 

  19. Figueiredo, M., Cunha, S., Martins, G., Freitas, J., Judas, F., Figueiredo, H.: Influence of hydrochloric acid concentration on the demineralization of cortical bone. Chem. Eng. Res. Des. 89(1), 116–124 (2011)

    Article  Google Scholar 

  20. Castro-Ceseña, A.B., Novitskaya, E.E., Chen, P.Y., Hirata, G.A., McKittrick, J.: Kinetic studies of bone demineralization at different HCl concentrations and temperatures. Mat. Sci. Eng. C 31(3), 523–530 (2011)

    Article  Google Scholar 

  21. Danilchenko, S.N., Moseke, C., Sukhodub, L.F., Sulkio-Cleff, B.: X-ray diffraction studies of bone apatite under acid demineralization. Cryst. Res. Technol. 39(1), 71–77 (2004)

    Article  Google Scholar 

  22. El-Bassyouni, G.T., Guirguis, O.W., Abdel-Fattah, W.I.: Morphological and macrostructural studies of dog cranial bone demineralized with different acids. Curr. Appl. Phys. 13(5), 864–874 (2013)

    Article  ADS  Google Scholar 

  23. Lewandrowski, K.U., Tomford, W.W., Michaud, N.A., Schomacker, K.T., Deutsch, T.F.: An electron microscopic study on the process of acid demineralization of cortical bone. Calcif. Tissue Int. 61(4), 294–297 (1997)

    Article  Google Scholar 

  24. Walsh, W.R., Christiansen, D.L.: Demineralized bone matrix as a template for mineral-organic composites. Biomaterials 16(18), 1363–1371 (1995)

    Article  Google Scholar 

  25. Teitelbaum, S.L.: Bone resorption by osteoclasts. Science 289(5484), 1504–1508 (2000)

    Article  ADS  Google Scholar 

  26. Akbay, A., Bozkurt, G., Ilgaz, O., Palaoglu, S., Akalan, N., Benzel, E.C.: A demineralized calf vertebra model as an alternative to classic osteoporotic vertebra models for pedicle screw pullout studies. Eur. Spine J. 17(3), 468–473 (2008)

    Article  Google Scholar 

  27. Lee, C.Y., Chan, S.H., Lai, H.Y., Lee, S.T.: A method to develop an in vitro osteoporosis model of porcine vertebrae: histological and biomechanical study: laboratory investigation. J. Neurosurg. Spine 14(6), 789–798 (2011)

    Article  Google Scholar 

  28. Ehrlich, H., Koutsoukos, P.G., Demadis, K.D., Pokrovsky, O.S.: Principles of demineralization: modern strategies for the isolation of organic frameworks: part II. Decalcification. Micron 40(2), 169–193 (2009)

    Article  Google Scholar 

  29. Bish, D.L., Post, J.E. (eds.): Modern powder diffraction. In: Ribbe, P.H. (ed.) Reviews in Mineralogy, 20. Mineralogical Society of America, Washington, DC (1989)

  30. Langford, J.I., Wilson, A.J.C.: Scherrer after sixty years: a survey and some new results in the determination of crystallite size. J. Appl. Crystallogr. 11(2), 102–113 (1978)

    Article  Google Scholar 

  31. Danilchenko, S.N., Kukharenko, O.G., Moseke, C., Protsenko, I.Y., Sukhodub, L.F., Sulkio-Cleff, B.: Determination of the bone mineral crystallite size and lattice strain from diffraction line broadening. Cryst. Res. Technol. 37(11), 1234–1240 (2002)

    Article  Google Scholar 

  32. Doi, Y., Moriwaki, Y., Aoba, T., Takahashi, J., Joshin, K.: ESR and IR studies of carbonate-containing hydroxyapatites. Calcif. Tissue Int. 34(1), 178–181 (1982)

    Article  Google Scholar 

  33. Rogers, K., Beckett, S., Kuhn, S., Chamberlain, A., Clement, J.: Contrasting the crystallinity indicators of heated and diagenetically altered bone mineral. Palaeogeogr. Palaeoclimatol. Palaeoecol. 296(1), 125–129 (2010)

    Article  Google Scholar 

  34. Pan, H., Tao, J., Yu, X., Fu, L., Zhang, J., Zeng, X., Tang, R.: Anisotropic demineralization and oriented assembly of hydroxyapatite crystals in enamel: smart structures of biominerals. J. Phys. Chem. B 112(24), 7162–7165 (2008)

    Article  Google Scholar 

  35. Tseng, W.J., Lin, C.C., Shen, P.W., Shen, P.: Directional/acidic dissolution kinetics of (OH, F, Cl)-bearing apatite. J. Biomed. Mater. Res. A 76(4), 753–764 (2006)

    Article  Google Scholar 

  36. Turunen, M.J., Kaspersen, J.D., Olsson, U., Guizar-Sicairos, M., Bech, M., Schaff, F., Isaksson, H.: Bone mineral crystal size and organization vary across mature rat bone cortex. J. Struct. Biol. 195(3), 337–344 (2016)

    Article  Google Scholar 

  37. Kaspersen, J.D., Turunen, M.J., Mathavan, N., Lages, S., Pedersen, J.S., Olsson, U., Isaksson, H.: Small-angle X-ray scattering demonstrates similar nanostructure in cortical bone from young adult animals of different species. Calcif. Tissue Int. 99, 76–87 (2016)

    Article  Google Scholar 

  38. Weiner, S., Wagner, H.D.: The material bone: structure-mechanical function relations. Annu. Rev. Mater. Sci. 28(1), 271–298 (1998)

    Article  ADS  Google Scholar 

  39. Baig, A.A., Fox, J.L., Young, R.A., Wang, Z., Hsu, J., Higuchi, W.I., Otsuka, M.: Relationships among carbonated apatite solubility, crystallite size, and microstrain parameters. Calcif. Tissue Int. 64(5), 437–449 (1999)

    Article  Google Scholar 

Download references

Acknowledgements

This work was partially supported by grants from the International Science & Technology Cooperation Program of China (ISTCP, NO. 2015DFR30940), Special Program from Chinese Academy of Science in Cooperation with Russia, Ukraine and the Republic of Belarus (2015, 2017) and the Introduced Intelligence project from the State Administration of Foreign Experts Affairs P.R. China (2016).

Author information

Authors and Affiliations

  1. Institute for Applied Physics, NAS of Ukraine, Sumy, Ukraine

    Sergei Danilchenko, Aleksei Kalinkevich, Mykhailo Zhovner & Vladimir Kuznetsov

  2. Institute of Modern Physics, CAS, Lanzhou, China

    He Li & Jufang Wang

Authors
  1. Sergei Danilchenko
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aleksei Kalinkevich
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mykhailo Zhovner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vladimir Kuznetsov
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. He Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jufang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Aleksei Kalinkevich.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Danilchenko, S., Kalinkevich, A., Zhovner, M. et al. Anisotropic aspects of solubility behavior in the demineralization of cortical bone revealed by XRD analysis. J Biol Phys 45, 77–88 (2019). https://doi.org/10.1007/s10867-018-9516-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10867-018-9516-5

Keywords

Search

Navigation