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The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth

  • Laboratory Investigations
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Summary

The calcification of connective tissues, including cartilage, is under the control of many interacting systems. Proteoglycans are thought to retard the deposition of hydroxyapatite crystals, and modification of the proteoglycans presumably facilitates mineralization in those tissues that are actively calcifying. The mechanism underlying these regulations remains speculative. This study investigates this question by comparing the inhibitory effectiveness of several macromolecules at neutral pH and approximately physiological ionic strengths. Inhibitors tested include bovine nasal proteoglycan monomer A1D1D1 and aggregate-containing A1 fractions, glycosaminoglycan chains (chondroitin 4-sulfate), and neutral dextran (as an uncharged analog). Hydroxyapatite growth was assessed either by measuring the time-dependent decreases in solution calcium and phosphate concentrations, or by determining utilization of hydroxyl ion in a pH-Stat. All species studied inhibit hydroxyapatite growth, and the extent of inhibition for each class is concentration-dependent. The proteoglycan aggregate-containing A1 fraction is more effective than the proteoglycan monomer at the same concentration, and the proteoglycan monomer is more effective than chondroitin 4-sulfate. Neutral dextran inhibits hydroxyapatite growth less effectively than proteoglycans. These results suggest that inhibition of hydroxyapatite growth by proteoglycans critically depends on both status (aggregate, monomer, etc.) and hydrodynamic size of this macromolecule, supporting the hypothesis that modification of proteoglycansin vivo functions to modulate the effectiveness of proteoglycans as a hydroxyapatite growth inhibitor.

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References

  1. Blumenthal NC, Posner AS, Silverman LD, Rosenberg LC (1979) Effect of proteoglycans on in vitro hydroxyapatite formation, Calcif Tissue Int 27:75–82

    CAS  PubMed  Google Scholar 

  2. Howell DS, Pita JC, Marquez JF, Gatter RA (1969) Demonstration of macromolecules inhibitors of calcification and nucleational factors in fluid from calcifying sites in cartilage, J Clin Invest 48:630–641

    Article  CAS  PubMed  Google Scholar 

  3. Cuervo LA, Pita JC, Howell DS (1973) Inhibition of calcium phosphate mineral growth by proteoglycan aggregate fractions in a synthetic lymph. Calcif Tissue Res 13:1–10

    Article  CAS  PubMed  Google Scholar 

  4. Howell DS, Pita JC (1976) Calcification of growth plate cartilage with specific reference to studies on micropuncture fluids. Clin Orthop Rel Res 118:208–229

    CAS  Google Scholar 

  5. Muir H, Hardingham TE (1975) Structure of proteoglycans. In: Whelan WJ (ed) Biochemistry of carbohydrates. Vol. 5. Butterworths/University Park Press, Baltimore, pp 153–221

    Google Scholar 

  6. Rosenberg L (1975) Structure of cartilage proteoglycans. In: Burleigh PMC, Poole AR (eds) Dynamics of connective tissue macromolecules. North-Holland, New York, pp 105–127

    Google Scholar 

  7. Muir H (1980) The chemistry of the ground substance of joint cartilage. In: Sokoloff L (ed) The joints and synovial fluid. Vol. 2. Academic Press, New York, pp 27–94

    Google Scholar 

  8. Buckwalter JA (1983) Proteoglycan structure in calcifying cartilage. Clin Orthop Rel Res 172:207–232

    CAS  Google Scholar 

  9. Matsuzawo VJ, Kotkos GA (1968) Evidence for changes in protein polysaccharide associated with the onset of calcification in cartilage. J Cell Biol 39:43–48

    Article  Google Scholar 

  10. deBernard B (1982) Glycoproteins in the local mechanism of calcification. Clin Orthop Rel Res 162:233–244

    CAS  Google Scholar 

  11. Lindenbaum A, Kuettner KE (1967) Mucopolysaccharides and mucoproteins of calf scapula. Calcif Tissue Res 1:153–165

    Article  CAS  PubMed  Google Scholar 

  12. Hirschman A, Dziewiatkowski DD (1966) Protein-polysaccharide loss during endochondral ossification: immunochemical evidence. Science 154:393–395

    CAS  PubMed  Google Scholar 

  13. Reddi AH, Hascall VC, Hascall GK (1978) Changes in proteoglycan types during matrix-induced cartilage and bone development. J Biol Chem 253 (7):2429–2436

    CAS  PubMed  Google Scholar 

  14. Brighton CT, Sugioka Y, Hunt RM (1973) Cytoplasmic structures of epiphyseal plate chondrocytes. J Bone Joint Surg (Am) 55-A:771–784

    Google Scholar 

  15. Kuettner KE, Eisenstein R, Sorgente N (1975) Lysozyme in calcifying tissues. Clin Orthop Rel Res 112:316–339

    CAS  Google Scholar 

  16. Granda JL, Posner AS (1971) Distribution of four hydrolases in the epiphyseal plate. Clin Orthop Rel Res 74:269–272

    CAS  Google Scholar 

  17. Rath NC, Hand AR, Reddi AH (1981) Activity and distribution of lysosomal enzymes during collagenous matrix-induced cartilage, bone and bone marrow development. Dev Biol 85:89–98

    Article  CAS  PubMed  Google Scholar 

  18. Thyberg J, Friberg U (1978) The lysosomal system in endochondral growth. Progress in histochemistry and cytochemistry 10(4):1–45

    CAS  PubMed  Google Scholar 

  19. Woessner JF, Howell DS (1983) Hydrolytic enzymes in cartilage. In: Maroudas A, Holborow EJ (eds) Studies in joint disease. Vol. 2. Pittman books Ltd. London, pp 106–152

    Google Scholar 

  20. Heinegård D (1972) Extraction, fractionation and characterization of proteoglycans from bovine nasal tracheal cartilage. Biochim Biophys Acta 85:181–192

    Google Scholar 

  21. Rosenberg L, Wolfenstein-Todel C, Margolis R, Pal S, Strider W (1976) Proteoglycans from bovine proximal humeral articular cartilage. J Biol Chem 251:6439–6444

    CAS  PubMed  Google Scholar 

  22. Bitter T, Muir HM (1962) A modified uronic acid and carbazole reaction. Anal Biochem 4:330–334

    Article  CAS  PubMed  Google Scholar 

  23. Dodgson KS, Price RG (1962) A note on the determination of the ester sulphate content of sulphated polysaccharides. Biochem J 84:106–110

    CAS  PubMed  Google Scholar 

  24. Posner AS (1969) Crystal chemistry of bone mineral. Physiol Rev 49(4):760–792

    CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  26. Crouch SR, Malmstadt HU (1967) A mechanistic investigation of molybdenum-blue method for determination of phosphate. Anal Chem 39:1084–1089

    Article  CAS  Google Scholar 

  27. Mitchell N, Shepard N, Harrod J (1982) The measurement of proteoglycan in the mineralizing region of the rat growth plate: an electron microscopic and x-ray microanalytical study. J Bone Joint Surg (Am) 64A:32–38

    Google Scholar 

  28. Kantor TG, Schubert M (1957) The difference in permeability of cartilage to cationic and anionic dyes. J Histochem Cytochem 5:28–32

    CAS  PubMed  Google Scholar 

  29. Rosenberg LC (1981) Proteoglycan aggregate structure in mineralizing cartilage. Orthop Trans 5:220

    Google Scholar 

  30. Fisher LW, Termine JD, Dejter SW, Whitson SW, Yanagishita MY, Kimura JH, Hascall VC, Kleinman HK, Hascall JR, Nilsson B (1983) Proteoglycans of developing bone. J Biol Chem 258:6588–6594

    CAS  PubMed  Google Scholar 

  31. Ruggeri A, Dell'orbo C, Ovacli D (1975) Electron microscopic visualization of proteoglycans with alcian blue. Histochem J 7:187–197

    Article  CAS  PubMed  Google Scholar 

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Chen, CC., Boskey, A.L. & Rosenberg, L.C. The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth. Calcif Tissue Int 36, 285–290 (1984). https://doi.org/10.1007/BF02405332

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