Calcified Tissue International

, Volume 93, Issue 4, pp 307–315 | Cite as

Biomineralization Mechanisms: A New Paradigm for Crystal Nucleation in Organic Matrices

Original Research


There is substantial practical interest in the mechanism by which the carbonated apatite of bone mineral can be initiated specifically in a matrix. The current literature is replete with studies aimed at mimicking the properties of vertebrate bone, teeth, and other hard tissues by creating organic matrices that can be mineralized in vitro and either functionally substitute for bone on a permanent basis or serve as a temporary structure that can be replaced by normal remodeling processes. A key element in this is mineralization of an implant with the matrix and mineral arranged in the proper orientations and relationships. This review examines the pathway to crystallization from a supersaturated calcium phosphate solution in vitro, focusing on the basic mechanistic questions concerning mineral nucleation and growth. Since bone and dentin mineral forms within collagenous matrices, we consider how the in vitro crystallization mechanisms might or might not be applicable to understanding the in vivo processes of biomineralization in bone and dentin. We propose that the pathway to crystallization from the calcium phosphate–supersaturated tissue fluids involves the formation of a dense liquid phase of first-layer bound-water hydrated calcium and phosphate ions in which the crystallization is nucleated. SIBLING proteins and their in vitro analogs, such as polyaspartic acids, have similar dense liquid first-layer bound-water surfaces which interact with the dense liquid calcium phosphate nucleation clusters and modulate the rate of crystallization within the bone and dentin collagen fibril matrix.


Biomineralization mechanism Matrix protein Dental matrix biology Dense liquid Hydration 


  1. 1.
    Veis A (2003) Mineralization in organic matrix frameworks. In: Dove PM, DeYoreo JJ, Weiner S (eds) Biomineralization. Reviews in mineralogy and geochemistry, vol 54. Mineralogical Society of America, Washington, DC, pp 249–289Google Scholar
  2. 2.
    Landis WJ, Song MJ, Leith A, McEwen L, McEwen B (1993) Mineral and organic matrix interaction in normally calcifying tendon visualized in three dimensions by high voltage electron microscopic tomography and graphic image reconstruction. J Struct Biol 110:39–54PubMedCrossRefGoogle Scholar
  3. 3.
    Orgel JPRO, San Antonio JD, Antipova O (2011) Molecular and structural mapping of collagen fibril interactions. Connect Tissue Res 52:2–17PubMedCrossRefGoogle Scholar
  4. 4.
    Silver FH, Landis WJ (2011) Deposition of apatite in mineralizing vertebrate extracellular matrices: a model of possible nucleation sites on type I collagen. Connect Tissue Res 52:242–254PubMedCrossRefGoogle Scholar
  5. 5.
    Wang Y, Azaïs T, Robin M, Vallée A, Catania C, Legriel P, Pehau-Arnaudet G, Babonneau F, Giraud-Guille M-M, Nassif N (2012) The predominant role of collagen in the nucleation, growth, structure and orientation of bone apatite. Nat Mater 11:724–733PubMedCrossRefGoogle Scholar
  6. 6.
    Veis A, Schlueter R (1963) Presence of phosphate-mediated cross-linkages in hard tissue. Collagens Nat 197:1204CrossRefGoogle Scholar
  7. 7.
    Veis A, Schlueter RJ (1964) The macromolecular organization of dentine matrix collagen I. Characterization of dentine collagen. Biochemistry 3:1650–1656PubMedCrossRefGoogle Scholar
  8. 8.
    Schlueter RJ, Veis A (1964) The macromolecular organization of dentine matrix collagen II. Periodate degradation and carbohydrate cross-linking. Biochemistry 3:1657–1665PubMedCrossRefGoogle Scholar
  9. 9.
    Veis A, Perry A (1967) The phosphoprotein of the dentin matrix. Biochemistry 6:2409–2416PubMedCrossRefGoogle Scholar
  10. 10.
    Veis A, Spector A, Carmichael DJ (1969) The organization and polymerization of bone and dentin collagen. Clin Orthop Relat Res 66:188–211PubMedCrossRefGoogle Scholar
  11. 11.
    Dimuzio MT, Veis A (1978) The biosynthesis of phosphophoryns and dentin collagen in the continuously erupting rat incisor. J Biol Chem 253:6845–6852PubMedGoogle Scholar
  12. 12.
    Dimuzio MT, Veis A (1978) Phosphophoryns—major non-collagenous proteins of the rat incisor dentin. Calcif Tissue Res 25:169–178PubMedCrossRefGoogle Scholar
  13. 13.
    Lee SL, Veis A, Glonek T (1977) Dentin phosphoprotein: an extracellular calcium binding protein. Biochemistry 16:2971–2979PubMedCrossRefGoogle Scholar
  14. 14.
    Veis A, Sharkey M, Dickson IR (1977) Non-collagenous proteins of bone and dentin extracellular matrix and their role in organized mineral deposition. In: Wasserman RH, Corradino E, Carafoli RH, Kretsinger RH, MacLennan DH, Siegel FL (eds) Calcium binding proteins and calcium function: proceedings of the international symposium on calcium-binding proteins and calcium function in health and disease. Elsevier, Amsterdam, North-Holland, pp 409–418Google Scholar
  15. 15.
    Lee SL, Veis A (1980) Studies on the structure and chemistry of dentin collagen–phosphophoryn covalent complexes. Calcif Tissue Res 31:123–134CrossRefGoogle Scholar
  16. 16.
    Weinstock M, Leblond CP (1973) Radioautographic visualization of the deposition of a phosphoprotein at the mineralization front in the dentin of the rat incisor. J Cell Biol 56(3):838–845PubMedCrossRefGoogle Scholar
  17. 17.
    Weinstock M, Leblond CP (1974) Synthesis, migration, and release of precursor collagen by odontoblasts as visualized by radioautography after (3H) proline administration. J Cell Biol 60(1):92–127PubMedCrossRefGoogle Scholar
  18. 18.
    Rabie AM, Veis A (1995) An immunocytochemical study of the routes of secretion of collagen and phosphophoryn from odontoblasts. Connect Tissue Res 31:197–209PubMedCrossRefGoogle Scholar
  19. 19.
    Fisher LW, Torchia DA, Fohr B, Young MF, Fedarko NS (2001) Flexible structures of SIBLING proteins, bone sialoprotein, and osteopontin. Biochem Biophys Res Commun 280:460PubMedCrossRefGoogle Scholar
  20. 20.
    Fisher LW, Fedarko NS (2003) Six genes expressed in bones and teeth encode the current members of the SIBLING family of proteins. Connect Tissue Res 44:33PubMedGoogle Scholar
  21. 21.
    Feenstra TP (1980) The initial stages in the formation of calcium and strontium phosphates from supersaturated solutions: a study of induction times. Dissertation, University of Utrecht, The Netherlands, pp 32–33Google Scholar
  22. 22.
    Schmelzer JW, Boltachev GS, Baidakov VG (2006) Classical and generalized Gibbs’ approaches and the work of critical cluster formation in nucleation theory. J Chem Phys 124:194503PubMedCrossRefGoogle Scholar
  23. 23.
    ten Wolde PR, Frenkel D (1997) Enhancement of crystal nucleation by critical density fluctuations. Science 277:1975–1978PubMedCrossRefGoogle Scholar
  24. 24.
    Talanquer V, Oxtoby DW (1998) Crystal nucleation in the presence of a metastable critical point. J Chem Phys 109:223–227CrossRefGoogle Scholar
  25. 25.
    Vekilov PG (2004) Dense liquid precursor for the nucleation of ordered solid phases from solution. Cryst Growth Des 4:671–685CrossRefGoogle Scholar
  26. 26.
    Vekilov PG (2010) Nucleation. Cryst Growth Des 10:5007–5019PubMedCrossRefGoogle Scholar
  27. 27.
    Wolf S, Leiterer J, Kappl M, Emmerling F, Tremel W (2008) Early homogenous amorphous precursor stages of calcium carbonate and subsequent crystal growth in levitated droplets. J Am Chem Soc 130:12342–12347PubMedCrossRefGoogle Scholar
  28. 28.
    Mahamid J, Sharir A, Addadi L, Weiner S (2008) Amorphous calcium phosphate is a major component of the forming fin bones of zebrafish: indications for an amorphous precursor phase. Proc Natl Acad Sci 105:12748–12753PubMedCrossRefGoogle Scholar
  29. 29.
    Beniash E, Metzler RA, Lam RS, Gilbert PU (2009) Transient amorphous calcium phosphate in forming enamel. J Struct Biol 166:133–143PubMedCrossRefGoogle Scholar
  30. 30.
    Posner AS (1969) Crystal chemistry of bone mineral. Physiol Rev 49:760–792PubMedGoogle Scholar
  31. 31.
    Posner AS, Betts F (1975) Synthetic amorphous calcium phosphate and its relation to bone mineral structure. Acc Chem Res 8:273–281CrossRefGoogle Scholar
  32. 32.
    Dey A, Bomans PHH, Muller FA, Will J, Frederik PM, de With G, Sommerdijk NAJM (2010) The role of prenucleation clusters in surface-induced calcium phosphate crystallization. Nat Mater 9:1010–1014PubMedCrossRefGoogle Scholar
  33. 33.
    Müller L, Müller F (2006) Preparation of SBF with different HCO3 content and its influence on the composition of biomimetic apatites. Acta Biomater 2:181–189PubMedCrossRefGoogle Scholar
  34. 34.
    Nudelman F, Pieterse K, George A, Bomans PHH, Friedrich H, Brylka LJ, Hilbers PAJ, de With G, Sommerdijk NAJM (2010) The role of collagen in bone apatite formation in the presence of hydroxyapatite nucleation inhibitors. Nat Mater 9:1004–1009PubMedCrossRefGoogle Scholar
  35. 35.
    Gebauer D, Völkel A, Cölfen H (2008) Stable prenucleation calcium carbonate clusters. Science 322:1816–1822CrossRefGoogle Scholar
  36. 36.
    Gower L (2008) Biomimetic model systems for investigating the amorphous precursor pathway and its role in biomineralization. Chem Rev 108:4551–4627PubMedCrossRefGoogle Scholar
  37. 37.
    Uversky VN (2002) What does it mean to be natively unfolded? Eur J Biochem 269:2–12PubMedCrossRefGoogle Scholar
  38. 38.
    Traub W, Jodaikin A, Arad T, Veis A, Sabsay B (1992) Dentin phosphophoryn binding to collagen fibrils. Matrix 12:197–201PubMedCrossRefGoogle Scholar
  39. 39.
    Dahl T, Sabsay B, Veis A (1998) Type I collagen–phosphophoryn interactions: specificity of the monomer–monomer binding. J Struct Biol 123:162–168PubMedCrossRefGoogle Scholar
  40. 40.
    Veis A, Dahl T, Sabsay B (2000) The specificity of phosphophoryn–collagen I interactions. In: Goldberg M, Robinson C, Boskey A (eds) Proceedings of the 6th international conference on the chemistry and biology of mineralized tissues, Vittel, France, November 1–6, 1998. Orthopaedic Research Society, Illinois, pp 169–173Google Scholar
  41. 41.
    Dahl T, Veis A (2003) Electrostatic interactions lead to the formation of asymmetric collagen–phosphophoryn aggregates. Connect Tissue Res 44(Suppl 1):206–213PubMedGoogle Scholar
  42. 42.
    He G, Ramachandran A, Dahl T, George S, Schultz D, Cookson D, Veis A, George A (2005) Phosphorylation of phosphophoryn is crucial for its function as a mediator of biomineralization. J Biol Chem 280:33109–33114PubMedCrossRefGoogle Scholar
  43. 43.
    Sfeir C, Lee D, Li J, Zhang X, Boskey AL, Kumta PN (2011) Expression of phosphophoryn is sufficient for the induction of matrix mineralization by mammalian cells. J Biol Chem 286:20228–20238PubMedCrossRefGoogle Scholar
  44. 44.
    Bonucci E (1967) Fine structure of early cartilage calcification. J Ultrastruct Res 20:33–50PubMedCrossRefGoogle Scholar
  45. 45.
    Bonucci E (2005) Calcified tissue: from microstructures to nanoparticles to chemistry. Eur J Histochem 49:1–10PubMedGoogle Scholar
  46. 46.
    Anderson HC, Matsuzawa T, Sajdera SW, Ali SY (1970) Membranous particles in calcifying cartilage matrix. Trans N Y Acad Sci 32:619–630PubMedCrossRefGoogle Scholar
  47. 47.
    Anderson HC (1967) Electron microscopic studies of induced cartilage development and calcification. J Cell Biol 35:81–101PubMedCrossRefGoogle Scholar
  48. 48.
    Lehninger A (1970) Mitochondria and calcium ion transport. Biochem J 119:128–138Google Scholar
  49. 49.
    Greenwalt JW, Rossi CS, Lehninger AL (1964) Effect of active accumulation of calcium and phosphate ions on the structure of rat liver mitochondria. J Cell Biol 23:21–38CrossRefGoogle Scholar
  50. 50.
    Boonrungsiman S, Gentleman E, Carzaniga R, Evans ND, McComb DW, Porter E, Stevens MM (2012) The role of intracellular calcium phosphate in osteoblast-mediated bone apatite formation. Proc Natl Acad Sci USA 109:14170–14175PubMedCrossRefGoogle Scholar
  51. 51.
    Fulton AB (1982) How crowded is the cytoplasm? Cell 30:345–347PubMedCrossRefGoogle Scholar
  52. 52.
    Fayer MD (2012) Dynamics of water interacting with interfaces, molecules and ions. Acc Chem Res 45:3–14PubMedCrossRefGoogle Scholar
  53. 53.
    Pavlov M, Seigbahn PEM, Sandstrom M (1998) Hydration of beryllium, magnesium, calcium and zinc ions using density functional theory. J Phys Chem A 102:219–228CrossRefGoogle Scholar
  54. 54.
    Richens DT (1997) The chemistry of aqua ions: synthesis, structure, and reactivity: a tour through the periodic table of elements. John Wiley & Sons Ltd, Chichester, EnglandGoogle Scholar
  55. 55.
    Pribil AB, Hofer TS, Randolf BR, Rode BM (2008) Structure and dynamics of phosphate ion in aqueous solution: an ab initio QMCF study. J Comput Chem 29:2330–2334PubMedCrossRefGoogle Scholar
  56. 56.
    George A, Bannon L, Sabsay B, Dillon JW, Malone J, Veis A, Jenkins NA, Gilbert DJ, Copeland NG (1996) The carboxyl-terminal domain of phosphophoryn contains unique extended triplet amino acid repeat sequences forming ordered carboxyl–phosphate interaction ridges that may be essential in the biomineralization process. J Biol Chem 271:32869–32873PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  1. 1.Department of Cell and Molecular BiologyFeinberg School of Medicine, Northwestern UniversityChicagoUSA

Personalised recommendations