The effect of phosphatidylserine onin vitro hydroxyapatite growth and proliferation


The acidic phospholipid phosphatidylserine (PS) has been reported to have variable effects onin vitro hydroxyapatite proliferation. PS promotesin vitro mineralization in systems in which calcium-PS-phosphate complexes are allowed to form, and inhibitsin vitro mineralization when incorporated into liposomes. To investigate these diverse effects, a Langmuir adsorption isotherm was used to determine the affinity of PS for hydroxyapatite crystals, based on binding of14C-PS to synthetic hydroxyapatite crystals of specific surface 54 m2/g. Using this model, PS was found to bind to hydroxyapatite crystals with an affinity comparable to that of the amino acid phosphoserine (K=3.33 ml/μmol). Coating the surface of hydroxyapatite seed crystals with PS reduced their rate of proliferation in a metastable calcium phosphate solution in which calcium-PS-phosphate complexes were previously shown to promote hydroxyapatite formation. The extent of inhibition of hydroxyapatite seeded growth was directly related to the proportion of the hydroxyapatite surface covered with PS. These data suggest that PS may have multiple effects on hydroxyapatite formationin situ, and that mineral-PS interactions can retard crystal proliferation.

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


  1. 1.

    Wuthier RE (1982) The role of phospholipid-calcium-phosphate in biological mineralization. In: Anghileri AJ, Tuffet-Anghileri AM (eds) The role of calcium in biological systems. I. CRC Press, Boca Raton, pp 41–70

    Google Scholar 

  2. 2.

    Boskey AL (1989) Phospholipids and calcification. In: Hukins DWL (ed) Calcified tissues. Macmillan Press, Ltd, London, pp 215–243

    Google Scholar 

  3. 3.

    Wuthier RE, Majeska RJ, Collins GM (1977) Biosynthesis of matrix vesicles in epiphyseal cartilage. I. In vivo incorporation of32P phosphate into phospholipids of chondrocyte, membrane and matrix vesicle fractions. Calcif Tissue Res 23:135–139

    PubMed  Article  CAS  Google Scholar 

  4. 4.

    Anderson HC (1980) Calcification processes. Pathol Ann 15:45–75

    Google Scholar 

  5. 5.

    Boskey AL, Posner AS (1982) Optimal conditions for the formation of Ca-phospholipid-phosphate complexes. Calcif Tissue Int 34:s1-s7

    PubMed  Article  Google Scholar 

  6. 6.

    Boskey AL, Posner AS (1977) The role of synthetic and bone extracted Ca-phospholipid-PO4 complexes in hydroxyapatite formation. Calcif Tissue Res 23:251–258

    PubMed  Article  CAS  Google Scholar 

  7. 7.

    Raggio CL, Boyan BD, Boskey AL (1986) In vivo hydroxyapatite formation induced by lipids. J Bone Min Res 1:409–415

    CAS  Article  Google Scholar 

  8. 8.

    Eanes ED, Hailer AW, Costa JL (1984) Calcium phosphate formation in aqueous suspensions of multilamellar liposomes. Calcif Tissue Int 36:421–430

    PubMed  Article  CAS  Google Scholar 

  9. 9.

    Eanes ED, Hailer AW (1987) Calcium phosphate precipitation in aqueous suspensions of phosphatidylserine-containing anionic liposomes. Calcif Tissue Int 40:43–48

    PubMed  Article  CAS  Google Scholar 

  10. 10.

    Heywood BR (1989) Calcium phosphate precipitation in liposomes. In: Recent advances in the study of dental calculus. IRL Press at Oxford University Press, London, pp 19–28

    Google Scholar 

  11. 11.

    Blumenthal NC, Betts F, Posner AS (1981) Formation and structure of Ca-deficient hydroxyapatite. Calcif Tissue Int 33:111–117

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    Willis JB (1960) The determination of metals in blood serum by atomic absorption spectroscopy. I. Calcium. Spectrochim Acta 16:259–272

    Article  CAS  Google Scholar 

  13. 13.

    Heinonen JL, Lahti RJ (1981) A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatases. Anal Biochem 113:313–317

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    Moreno EC, Kresak M, Hay DL (1982) Adsorption thermodynamics of acidic proline-rich human salivary proteins onto calcium apatites. J Biol Chem 257:2981–2989

    PubMed  CAS  Google Scholar 

  15. 15.

    Aoba T, Moreno EC (1985) Adsorption of phosphoserine onto hydroxyapatite and its inhibitory activity on crystal growth. J Colloid Interface Sci 106:110–121

    Article  CAS  Google Scholar 

  16. 16.

    Fiske CV, Subbarow Y (1925) The colorimetric determination of phosphorus. J Phys Chem 66:375–400

    CAS  Google Scholar 

  17. 17.

    Boskey AL, Wians FH, Hauschka PV (1985) The effect of osteocalcin on in vitro lipid-induced hydroxyapatite formation and seeded hydroxyapatite growth. Calcif Tissue Int 37:57–62

    PubMed  CAS  Google Scholar 

  18. 18.

    Aoba T, Moreno EC, Hay DI (1984) Inhibition of apatite crystal growth by the amino-terminal segment of human salivary acidic proline-rich proteins. Calcif Tissue Int 36:651–658

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    Meyers HM (1987) Structure-activity relationships (SAR) of hydroxyapatite-binding molecules. Calcif Tissue Int 40:344–348

    Article  Google Scholar 

  20. 20.

    Odutuga AA, Prout RES, Hoare RJ (1975) Hydroxyapatite precipitation in vitro by lipids extracted from mammalian tissues. Arch Oral Biol 20:311–315

    PubMed  Article  Google Scholar 

  21. 21.

    Vogel JJ, Boyan-Salyers BD (1976) Acidic lipids associated with the local mechanism of calcification. Clin Orthop 118:230–241

    CAS  Google Scholar 

  22. 22.

    Boyan B, Boskey AL (1984) Co-isolation of proteolipids and calcium-phospholipid-phosphate complexes. Calcif Tissue Int 36:214–218

    PubMed  Article  CAS  Google Scholar 

  23. 23.

    Boskey AL, Reddi AH (1983) Changes in lipids during matrix-induced endochondral bone formation. Calcif Tissue Int 35:549–554

    PubMed  Article  CAS  Google Scholar 

  24. 24.

    Boyan BD, Howell DS, Pita JC, Blanco L, Cieslak S (1988) Characterization of a calcification induction factor in epiphyseal cartilage extracellular fluid. Bone 9:185–194

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    Jung A, Bisaz S, Fleisch H (1973) The binding of pyrophosphate and two diphosphonates by hydroxyapatite crystals. Calcif Tissue Res 11:269–280

    PubMed  Article  CAS  Google Scholar 

  26. 26.

    Fleisch H, Bisaz S (1962) Isolation from urine of pyrophosphate, a calcification inhibitor. Am J Physiol 203:671–675

    PubMed  CAS  Google Scholar 

  27. 27.

    Francis MD, Russell RGG, Fleisch H (1969) Diphosphonates inhibit formation of calcium phosphate crystals in vitro and pathologic calcifications in vivo. Science 165:1264–1266

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    Boskey AL, Maresca M, Doty S, Sabsay B, Veis A (1990) Concentration-dependent effects of dentin phosphophoryn in the regulation of in vitro hydroxyapatite formation and growth. Bone Miner 10:55–65

    Article  Google Scholar 

Download references

Author information



Corresponding author

Correspondence to Adele L. Boskey.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Boskey, A.L., Dick, B.L. The effect of phosphatidylserine onin vitro hydroxyapatite growth and proliferation. Calcif Tissue Int 49, 193–196 (1991).

Download citation

Key words

  • Hydroxyapatite
  • Phosphatidylserine
  • Adsorption
  • Acidic phospholipids
  • Mineralization