Skip to main content
SpringerLink
Log in

Partition of inorganic ions and phospholipids in isolated cell, membrane and matrix vesicle fractions: Evidence for Ca-Pi-acidic phospholipid complexes

  • Published:
Calcified Tissue Research Aims and scope Submit manuscript

Summary

Electrolytes and phospholipids of cartilage fractions were partitioned by extraction with organic and aqueous solvents into six solubility groups: Electrolytes I, II and III, and Lipids I, II and III.

Of the total Ca, only 4% was water soluble (Electrolytes I); 4–12% was complexed with lipids (Electrolytes II); while the majority (84–92%) was insoluble (Electrolytes III). In contrast, nearly half of the Mg and Pi were water soluble. Of theneutral phospholipid, 95% was not complexed with mineral ions (Lipids I), but 30–45% of theacidic phospholipid was (Lipids II). Ca/Pi ratios were extremely low in the water-soluble phase, but were in the range of amorphous calcium phosphate (ACP) in the insoluble. Molar ratios of the lipid-mineral complex were: Ca∶Mg∶Pi:acidic phospholipid, 4∶3∶2∶2. Mg/Ca ratios in the soluble fraction were high (5.5–8.9), sufficient to stabilize ACP.

Kinetic studies revealed rapid turnover of soluble Ca, insoluble turning over much more slowly. Labeling of lipid-complexed Ca was rapid in cells, but occurred later in matrix vesicles, suggesting transfer. While lipid-Ca-Pi complexes can nucleate apatite in vitro, those present in vivo inside matrix vesicles apparently do not because of the excess Mg. We conclude therefore, that in vesicle-mediated calcification, lysis of the membrane may be essential to allow release of internal Mg.

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

Similar content being viewed by others

References

  1. Ali, S.Y.: Analysis of matrix vesicles and their role in calcification of epiphyseal cartilage. Fed. Proc.35, 135–142 (1976)

    PubMed  Google Scholar 

  2. Anderson, H.C.: Vesicles associated with calcification in the matrix of epiphyseal cartilage. J. Cell Biol.41, 59–72 (1969)

    PubMed  Google Scholar 

  3. Anderson, H.C.: Calcium-accumulating vesicles in the intercellular matrix of bone. In: Hard tissue growth, repair and remineralization, p. 213–246. Amsterdam: CIBA Foundation Symposium II, 1973.

    Google Scholar 

  4. Anghileri, L.J.: Calcium binding to phospholipids from experimental tumors. Z. Krebsforsch.78, 337–344 (1972)

    Google Scholar 

  5. Anghileri, L.J., Dermietzel, R.: Calcium-phosphate-phospholipids complexes in experimental tumors: their possible relationship to tumor calcification. Z. Krebsforsch.79, 148–156 (1973)

    Google Scholar 

  6. Bernard, G.W., Pease, D.C.: An electron microscopic study of initial intramembranous osteogenesis. Am. J. Anat.125, 271–290 (1969)

    PubMed  Google Scholar 

  7. Bonucci, E.: Fine structure of early cartilage calcification. J. Ultrastruct. Res.20, 33–50 (1967)

    PubMed  Google Scholar 

  8. Bonucci, E.: Fine structure and histochemistry of “calcifying globules” in epiphyseal cartilage. Z. Zellforsch.103, 192–217 (1970)

    PubMed  Google Scholar 

  9. Bonucci, E., Dearden, L.C.: Matrix vesicles in aging cartilage. Fed. Proc.35, 163–168 (1976)

    PubMed  Google Scholar 

  10. Boskey, A.L., Posner, A.S.: Magnesium stabilization of amorphous calcium phosphate: A kinetic study. Mat. Res. Bull.9, 907–916 (1974)

    Google Scholar 

  11. Boskey, A.L., Posner, A.S.: Extraction of calcium-phospholipid-phosphate complex from bone. Calcif. Tiss. Res.19, 273–284 (1976)

    Google Scholar 

  12. Boskey, A.L., Posner, A.S.: The role of synthetic and bone extracted Ca-phospholipid-PO4 complexes on hydroxyapatite formation. Calcif. Tiss. Res. (In Press)

  13. Cotmore, J.M., Nichols, G., Wuthier, R.E.: Phospholipid-calcium-phosphate complex: Enhanced calcium migration in the presence of phosphate. Science172, 1339–1341 (1971)

    PubMed  Google Scholar 

  14. Eisenberg, E., Wuthier, R.E., Frank, R.B., Irving, J.T.: Time study of in vivo incorporation of32P orthophosphate into phospholipids of chicken epiphyseal tissues. Calcif. Tiss. Res.6, 32–48 (1970)

    Google Scholar 

  15. Eisemann, D.R., Glick, P.L.: Ultrastructure of initial crystal formation in dentin. J. Ultrastruct. Res.41, 18–28 (1972)

    PubMed  Google Scholar 

  16. Ennever, J., Vogel, J.J., Rider, L.J., Boyan-Salyers, B.: Nucleation of microbiological calcification by proteolipid. Proc. Soc. exp. Biol. (N.Y.)152, 148–150 (1976)

    Google Scholar 

  17. Felix, R., Fleisch, H.: Pyrophosphatase and ATPase of isolated cartilage matrix vesicles. Calcif. Tiss. Res.22, 1–7 (1976)

    Google Scholar 

  18. Folch, J., Lees, M., Sloane-Stanley, G.H.: A simple method for the isolation and purification of total lipid from animal tissues. J. Biol. Chem.226, 497–509 (1957)

    PubMed  Google Scholar 

  19. Majeska, R.J., Wuthier, R.E.: Studies on matrix vesicles isolated from chick epiphyseal cartilage: Association of pyrophosphatase and ATPase activities with alkaline phosphatase. Biochim. Biophys. Acta (Amst.)391, 51–60 (1975)

    Google Scholar 

  20. Majeska, R.J., Wuthier, R.E.: Localization of matrix vesicle phospholipids by labeling with trinitrobenzenesulfonate (TNBS). Intern. Assn. Dental Res., Preprinted Abstracts, 54th General Session, Abstract No. 205 (1976)

  21. Mech, J.J.: Metabolism of45Ca in density fractions of lightly and heavily mineralized growth cartilage of pigs and chicks. M.S. Thesis, University of Vermont, Burlington, p. 22 (1974)

    Google Scholar 

  22. Rouser, G., Siakotos, A.N., Fleisher, S.: Quantitative analysis of phospholipids by thin-layer chormatography and phosphate analysis of spots. Lipids1, 85–86 (1966)

    Google Scholar 

  23. Termine, J.D., Peckauskas, R.A., Posner, A.S.: Calcium phosphate formation in vitro II. Effect of environment on amorphous-crystalline transformation. Arch. Biochem. Biophys.140, 318–325 (1970)

    PubMed  Google Scholar 

  24. Vogel, J.J., Boyan-Salyers, B.D.: Acidic lipids associated with the local mechanism of calcification. Clin. Orthop. Rel. Res.118, 230–241 (1976)

    Google Scholar 

  25. Wuthier, R.E.: Lipids of mineralizing epiphyseal tissues in the bovine fetus. J. Lipid. Res.9, 68–78 (1968)

    PubMed  Google Scholar 

  26. Wuthier, R.E.: Zonal analysis of phospholipids in the epiphyseal cartilage and bone of normal and rachitic chickens and pigs. Calcif. Tiss. Res.8, 36–53 (1971)

    Google Scholar 

  27. Wuthier, R.E.: The role of phospholipids in biological calcification. Distribution of phospholipase activity in calcifying epiphyseal cartilage. Clin. Orthop. Rel. Res.90, 191–200 (1973)

    Google Scholar 

  28. Wuthier, R.E.: Lipid composition of isolated epiphyseal cartilage cells, membranes and matrix vesicles. Biochim. Biophys. Acta (Amst.)409, 128–143 (1975)

    Google Scholar 

  29. Wuthier, R.E.: Electrolytes of isolated epiphyseal chondrocytes, matrix vesicles and extracellular fluid. Calcif. Tiss. Res.23, 125–133 (1977)

    Google Scholar 

  30. Wuthier, R.E., Eanes, E.D.: Effect of phospholipids on the transformation of amorphous calcium phosphate to hydroxyapatite in vitro. Calcif. Tiss. Res.19, 197–210 (1975)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemistry, The University of South Carolina, School of Medicine, 29208, Columbia, South Carolina, USA

    R. E. Wuthier & S. T. Gore

Authors
  1. R. E. Wuthier
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. T. Gore
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wuthier, R.E., Gore, S.T. Partition of inorganic ions and phospholipids in isolated cell, membrane and matrix vesicle fractions: Evidence for Ca-Pi-acidic phospholipid complexes. Calc. Tis Res. 24, 163–171 (1977). https://doi.org/10.1007/BF02223311

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02223311

Key words

Search

Navigation