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The Effect of Seed Crystals of Hydroxyapatite and Brushite on the Crystallization of Calcium Oxalate in Undiluted Human Urine In Vitro: Implications for Urinary Stone Pathogenesis

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Abstract

Background

The aim of this study was to determine whether crystals of hydroxyapatite (HA) or brushite (BR) formed in urine promote the epitaxial deposition of calcium oxalate (CaOx) from undiluted human urine in vitro and thereby explain the occurrence of phosphate in the core of urinary stones consisting predominantly of CaOx.

Materials and Methods

Crystals of HA, BR, and CaOx were generated from human urine and their identity confirmed by X-ray analysis. Standard quantities of each crystal were then added to separate aliquots of pooled undiluted human urine and CaOx crystallization was induced by the addition of identical loads of sodium oxalate. Crystallization was monitored by Coulter Counter and 14C-oxalate analysis and the precipitated crystals were examined by scanning electron microscopy.

Results

In comparison with the control to which no seeds were added, addition of CaOx crystals increased the deposition of 14C-oxalate by 23%. On the other hand, seeds of HA and BR had no effect. These findings were supported by Coulter Counter analysis, which showed that the average modal sizes of crystal particles precipitated in the presence of HA and BR seeds were indistinguishable from those in the control, whereas those deposited in the presence of CaOx were significantly larger. Scanning electron microscopy confirmed these results, demonstrating that large aggregates of CaOx dihydrates were formed in the presence of CaOx seeds, whereas BR and to a lesser extent HA seeds were scattered free on the filtration membrane and attached like barnacles on the surface of the freshly precipitated CaOx crystals.

Conclusion

Seed crystals of HA or BR do not promote CaOx deposition in urine in vitro and are therefore unlikely to influence CaOx crystal formation under physiologic conditions. However, binding of HA and BR crystals to, and their subsequent enclosure within, actively growing CaOx crystals might occur in vivo, thereby explaining the occurrence of mixed oxalate/phosphate stones.

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References

  1. Sierakowski R, Finlayson B, Landes RR, Finlayson CD, Sierakowski N. (1978) The frequency of urolithiasis in hospital discharge diagnoses in the United States. Invest. Urol. 15: 438–441.

    PubMed  CAS  Google Scholar 

  2. Robertson WG, Peacock M, Heyburn PJ, Hanes FA. (1980) Epidemiological risk factors in calcium stone disease. Scand. J. Urol. Nephrol. 53 (suppl): 15–28.

    CAS  Google Scholar 

  3. Soucie JM, Thun MJ, Coates RJ, McClellan W, Austin H. (1994) Demographic and geographic variability of kidney stones in the United States. Kidney Int. 46: 893–899.

    Article  CAS  PubMed  Google Scholar 

  4. Robertson WG. (2001) The changing pattern of urolithiasis in the UK and its causes. In Kok DJ, Romijn HC, Verhagen PCMS, Verkoelen CF (eds). Eurolithiasis. Maastricht, The Netherlands: Shaker Publishing; 9–11.

    Google Scholar 

  5. Clark JY, Thompson IM, Optenberg SA. (1995) Economic impact of urolithiasis in the United States. J. Urol. 154: 2020–2024.

    Article  CAS  PubMed  Google Scholar 

  6. Prien EL, Prien EL Jr. (1968) Composition and structure of urinary stone. Am. J. Med. 45: 654–672.

    Article  CAS  PubMed  Google Scholar 

  7. Elliot JS. (1973) Structure and composition of urinary calculi. J. Urol. 109: 82–83.

    Article  CAS  PubMed  Google Scholar 

  8. Modlin M. (1967) The aetiology of renal stones: a new concept arising from studies in a stone-free population. Ann. R. Coll. Surg. Engl. (Lond.) 40: 155–178.

    CAS  Google Scholar 

  9. Boyce WH, King JS Jr. (1963) Present concepts concerning the origin of matrix and stones. Ann. N. Y. Acad. Sci. 104: 563–578.

    Article  CAS  PubMed  Google Scholar 

  10. Abraham PA, Smith CL. (1987) Evaluation of factors involved in calcium stone formation. Miner. Electrolyte Metab. 13: 201–208.

    PubMed  CAS  Google Scholar 

  11. Royer L. (1928) Experimental research on parallel growth on mutual orientation of crystals of different species. Bull. Soc. Fr. Mineral 51: 7–159.

    CAS  Google Scholar 

  12. Lonsdale K. (1968) Epitaxy as a growth factor in urinary calculi and gallstones. Nature 217: 5658.

    Article  Google Scholar 

  13. Lonsdale K. (1968) Human stones: limited studies give some details of composition, rates of growth, distribution, and possible causes. Science 159: 1199–1207.

    Article  CAS  PubMed  Google Scholar 

  14. Robertson WG, Peacock M, Nordin BEC. (1968) Activity products in stone-forming and non-stone-forming urine. Clin. Sci. 34: 579–594.

    PubMed  CAS  Google Scholar 

  15. Pak CYC, Holt K. (1976) Nucleation and growth of brushite and calcium oxalate in urine of stone formers. Metabolism 25: 665–673.

    Article  CAS  PubMed  Google Scholar 

  16. Smith LH, Werness PG. (1983) Hydroxyapatite—the forgotten crystal in calcium urolithiasis. Trans. Am. Clin. Climatol. Assoc. 95: 183–190.

    Google Scholar 

  17. Berland Y, Boistelle R, Olmer M. (1990) Urinary supersaturation with respect to brushite in patients suffering from calcium oxalate lithiasis. Nephrol. Dial. Transplant. 5: 179–184.

    Article  CAS  PubMed  Google Scholar 

  18. Hallson PC, Rose GA. (1976) Crystalluria in normal subjects and in stone formers with and without thiazide and cellulose phosphate treatment. Br. J. Urol. 48: 515–524.

    Article  CAS  PubMed  Google Scholar 

  19. Werness PG, Wilson JWL, Smith LH. (1984) Hydroxyapatite and its role in calcium urolithiasis. In Ryall RL, Brockis JG, Marshall VR, Finlayson B (eds). Urinary Stones. Melbourne, London, New York: Churchill Livingstone; 273–277.

    Google Scholar 

  20. Tiselius HG. (1992) Recurrent stone formation in patients treated with ESWL. J. Stone Dis. 4: 152–157.

    Google Scholar 

  21. Tiselius HG, Larsson L. (1993) Calcium phosphate: an important crystal phase in patients with recurrent calcium stone formation? Urol. Res. 21: 175–180.

    Article  CAS  PubMed  Google Scholar 

  22. Pak CYC, Hayashi Y, Arnold LH. (1976) Heterogeneous nucleation with urate, calcium phosphate and calcium oxalate. Proc. Soc. Exp. Biol. Med. 153: 83–87.

    Article  CAS  PubMed  Google Scholar 

  23. Meyer JL, Bergert JH, Smith LH. (1975) Epitaxial relationships in urolithiasis: the calcium oxalate monohydratehydroxyapatite system. Clin. Sci. Mol. Med. 49: 369–374.

    PubMed  CAS  Google Scholar 

  24. Meyer JL, Bergert JH, Smith LH. (1977) Epitaxial relationships in urolithiasis: the brushite-whewellite system. Clin. Sci. Mol. Med. 52: 143–148.

    PubMed  CAS  Google Scholar 

  25. Koutsoukos PG, Sheehan ME, Nancollas GH. (1981) Epitaxial considerations in urinary stone formation. II. The oxalatephosphate system. Invest. Urol. 18: 358–363.

    PubMed  CAS  Google Scholar 

  26. Berg C, Tiselius HG. (1989) The effects of citrate on hydroxyapatite induced calcium oxalate crystallization and on the formation of calcium phosphate crystals. Urol. Res. 17: 167–172.

    Article  CAS  PubMed  Google Scholar 

  27. Baumann JM, Ackermann D, Affolter B. (1989) The influence of hydroxyapatite and pyrophosphate on the formation product of calcium oxalate at different pHs. Urol. Res. 17: 153–155.

    PubMed  CAS  Google Scholar 

  28. Mandel NS, Mandel GS. (1981) Epitaxis between stoneforming crystals at atomic level. In Smith LH, Robertson WG, Finlayson B (eds). Urolithiasis: Clinical and Basic Research. New York: Plenum; 469–480.

    Chapter  Google Scholar 

  29. Ebrahimpour A, Perez L, Nancollas GH. (1991) Induced crystal growth of calcium oxalate monohydrate at hydroxyapatite surfaces. The influence of human serum albumin, citrate and magnesium. Langmuir 7: 577–583.

    Article  CAS  Google Scholar 

  30. Achilles W, Jockel U, Schaper A, Burk B, Riedmiller H. (1995) In vitro formation of “urinary stones”: generation of spherulites of calcium phosphate in gel and overgrowth with calcium oxalate using a new flow model of crystallization. Scanning Microsc. 9: 577–586.

    PubMed  CAS  Google Scholar 

  31. Coe FL, Favus MJ, Pak CYC, Preminger GL. (1996) Kidney Stones: Medical and Surgical Management. Boston: Raven Press.

    Google Scholar 

  32. Hojgaard I, Fornander A-M, Nilsson M-A, Tiselius HG. (1998) The influence of hydroxyapatite seed on the crystallization induced by volume reduction of solutions with an ion composition corresponding to that in the distal tubule at different pH levels. Scand. J. Urol. Nephrol. 32: 311–319.

    Article  CAS  PubMed  Google Scholar 

  33. Beshensky AM, Wesson JA, Worcester EM, Snyder C, Kleinman JG. (2000) Urinary protein effects on heterogeneous nucleation of calcium oxalate on hydroxyapatite. In Rodgers AL, Hibbert BE, Hess B, Khan SR, Preminger GM (eds). Urolithiasis 2000. Cape Town, South Africa: University of Cape Town, Rondebosch; 116–118.

    Google Scholar 

  34. Stapleton AMF, Dawson CJ, Grover PK, et al. (1996) Further evidence linking urolithiasis and blood coagulation: urinary prothrombin fragment 1 is present in stone matrix. Kidney Int. 49: 880–888.

    Article  CAS  PubMed  Google Scholar 

  35. Nishio S, Hatanaka M, Takeda H, et al. (2000) Calcium phosphate crystal-associated proteins: alpha 2-HS-glycoprotein, prothrombin F1, and osteopontin. Mol. Urol. 4: 383–389.

    PubMed  CAS  Google Scholar 

  36. Doyle IR, Ryall RL, Marshall VR. (1991) Inclusion of proteins into calcium oxalate crystals precipitated from human urine: a highly selective phenomenon. Clin. Chem. 37: 1589–1594.

    PubMed  CAS  Google Scholar 

  37. Self PG, Pickering JG, Raven MD, Riley GG, Rosser H, Stone PA. (1987) Procedures for the control and manipulation of X-ray powder diffraction data from remote locations based on a Phillips PW1710 diffraction control system. I. Automatic control of the diffractometer—user’s manual. Glen Osmond, South Australia: CSIRO Division of Soils Technical Memo No. 75/1987.

    Google Scholar 

  38. Raven MD, Self PG. (1987) Procedures for the control and manipulation of X-ray powder diffraction data from remote locations based on a Phillips PW1710 diffraction control system. II. Handling procedures for data from the PW1710. Version 1.00. Glen Osmond, South Australia: CSIRO Division of Soils Technical Memo No. 91/1987.

    Google Scholar 

  39. Doyle IR, Ryall RL, Marshall VR. (1989) The effect of low-speed centrifugation and millipore filtration on the urinary protein content. In Walker VR, Sutton RAL, Cameron ECB, Pak CYC, Robertson WG (eds). Urolithiasis. New York: Plenum; 593–594.

    Chapter  Google Scholar 

  40. Ryall RL, Harnett RM, Hibberd CM, Edyvane KA, Marshall VR. (1991) Effects of chondroitin sulphate, human serum albumin and Tamm-Horsfall mucoprotein on calcium oxalate crystallization in undiluted human urine. Urol. Res. 19: 181–188.

    Article  CAS  PubMed  Google Scholar 

  41. National Institute of Health Consensus Development Conference Statement. (1989) Prevention and treatment of kidney stones. J. Urol. 141: 804–808.

    Article  Google Scholar 

  42. Ackermann D, Baumann JM. (1987) Chemical factors governing the state of saturation towards brushite and whewellite in urine of calcium stone formers. Urol. Res. 15: 63–65.

    Article  CAS  PubMed  Google Scholar 

  43. Robertson WG, Peacock M, Nordin BEC. (1969) Calcium crystalluria in recurrent renal-stone formers. Lancet 5: 21–24.

    Article  Google Scholar 

  44. Strates BS, Neuman WF, Levinskas GJ. (1957) The solubility of bone mineral. II. Precipitation of near neutral solutions of calcium and phosphate. J. Phys. Chem. 61: 279–282.

    Article  CAS  Google Scholar 

  45. Pak CYC, Eanes ED, Ruskin B. (1971) Spontaneous precipitation of brushite in urine: evidence that brushite is the nidus of renal stones originating as calcium phosphate. Proc. Natl. Acad. Sci. U.S.A. 68: 1456–1460.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Pak CYC. (1969) Physicochemical basis for formation of renal stones of calcium phosphate origin: calculation of the degree of saturation of urine with respect to brushite. J. Clin. Invest. 48: 1914–1922.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Pak CYC. (1981) Potential etiological role for brushite in the formation of calcium (renal) stones. J. Crystal Growth 53: 202–208.

    Article  CAS  Google Scholar 

  48. Pak CYC, Skinner HCW. (1968) Ionic interaction with bone mineral. IV. Varying affinity of synthetic calcium phosphates for Ca2+. Biochem. Biophys. Acta 165: 274–282.

    Article  CAS  PubMed  Google Scholar 

  49. Ryall RL. (1996) Glycosaminoglycans, proteins, and stone formation: adult themes and child’s play. Pediatr. Nephrol. 10: 656–666.

    Article  CAS  PubMed  Google Scholar 

  50. Worcester EM, Blumenthal SS, Beshensky AM, Lewand DL. (1992) The calcium oxalate growth inhibitor protein produced by mouse kidney cortical cells in culture is osteopontin. J. Bone Miner. Res. 7: 1029–1036.

    Article  CAS  PubMed  Google Scholar 

  51. Worcester EM, Beshensky AM. (1995) Osteopontin inhibits nucleation of calcium oxalate crystals. Ann. N. Y. Acad. Sci. 760: 375–377.

    Article  CAS  PubMed  Google Scholar 

  52. Grases F, Conte A, Gil JJ. (1988) Simple method for the study of heterogeneous nucleation in calcium oxalate urolithiasis. Br. J. Urol. 61: 468–473.

    Article  CAS  PubMed  Google Scholar 

  53. Grover PK, Ryall RL. (1997) Effect of seed crystals of uric acid and monosodium urate on the crystallization of calcium oxalate in undiluted human urine in vitro. Clin. Sci. 92: 205–213.

    Article  CAS  PubMed  Google Scholar 

  54. Khan SR. (1997) Calcium phosphate/calcium oxalate crystal association in urinary stones: implications for heterogeneous nucleation of calcium oxalate. J. Urol. 157: 376–383.

    Article  CAS  PubMed  Google Scholar 

  55. Burns JR, Finlayson B. (1980) The effect of seed crystals on calcium oxalate nucleation. Invest. Urol. 18: 133–136.

    PubMed  CAS  Google Scholar 

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Acknowledgments

The authors are indebted to Dr. A. Milnes, CSIRO, Division of Soils, Glen Osmond, South Australia, for performing the X-ray powder diffraction analyses of the seed crystals. Sincere thanks are also extended to Professor C.Y.C. Pak (University of Texas Southern Medical Center at Dallas, Dallas, Texas, USA) for providing the brushite seed crystals. This work was supported by grant 980366 from the National Health and Medical Research Council of Australia and grants from the Research Foundation of the Urological Society of Australasia, Flinders University of South Australia and Flinders 2000.

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  1. Dong-Sun Kim

    Present address: Department of Urology, Korea University Guro Hospital #80, Guro-Dong, Guro-Gu, Seoul, Korea

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  1. Department of Surgery, Flinders University School of Medicine, Flinders Medical Centre, Bedford Park, South Australia, 5042, Australia

    Phulwinder K. Grover, Dong-Sun Kim & Rosemary Lyons Ryall

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Grover, P.K., Kim, DS. & Ryall, R.L. The Effect of Seed Crystals of Hydroxyapatite and Brushite on the Crystallization of Calcium Oxalate in Undiluted Human Urine In Vitro: Implications for Urinary Stone Pathogenesis. Mol Med 8, 200–209 (2002). https://doi.org/10.1007/BF03402012

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