Skip to main content

Advertisement

SpringerLink
Log in

Mitigation of Ectopic Calcification in Osteopontin-Deficient Mice by Exogenous Osteopontin

  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

Ectopic calcification is a major cause of bioprosthetic heart valve failure. New therapeutic opportunities are offered by the growing understanding that ectopic calcification is an actively regulated process involving several key gene products. One of these products, osteopontin (OPN), is a glycosylated phosphoprotein previously shown to inhibit apatite crystal formation, induce carbonic anhydrase II, and promote mineral resorption. In this study, OPN-deficient mice (OPN−/−) were utilized as an in vivo model to stimulate the ectopic calcification of glutaraldehyde-fixed bovine pericardium (GFBP) tissue and to examine OPN delivery and structure-function relationships with respect to its anti-calcific activity. Significant calcification of GFBP tissue was obtained within 7 days of subcutaneous implantation in OPN−/− mice. Direct rescue of the calcification phenotype was achieved by the administration of exogenous recombinant rat, histidine-fused OPN (rat His-OPN) to the implant site via soluble injection (up to 72% mitigation achieved) or adsorption onto the implant materials (up to 91% mitigation achieved). Effects were specific, since neither fibronectin nor polyhistidine alone could mitigate calcification of GFBP. The maximum anti-calcific effect was achieved only when rat His-OPN was adequately phosphorylated and contained a functional arginine-glycine-aspartate (RGD) cell adhesive domain. Furthermore, CAII levels in host cells surrounding GFBP were greatest when phosphorylated, RGD-containing rat His-OPN was adsorbed. These data suggest that both physical inhibition, mediated by phosphorylation sites in OPN, as well as the induction of CAII and mineral regression, mediated by the RGD domain, contribute to the unique ability of OPN to mitigate ectopic calcification of bioprosthetic valve tissue.

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. FJ Schoen RJ Levy (1999) ArticleTitleFounder’s Award, 25th Annual Meeting of the Society for Biomaterials, Providence, RI, April 28–May 2, 1999. Tissue heart valves: current challenges and future research perspectives J Biomed Mater Res 47 439–465 Occurrence Handle1:CAS:528:DyaK1MXmvVSks7g%3D Occurrence Handle10497280

    CAS  PubMed  Google Scholar 

  2. CM Giachelli (1999) ArticleTitleEctopic calcification: gathering hard facts about soft tissue mineralization Am J Pathol 154 671–675 Occurrence Handle1:STN:280:DyaK1M7otVOmsQ%3D%3D Occurrence Handle10079244 Occurrence Handle1866412

    CAS  PubMed  PubMed Central  Google Scholar 

  3. SA Steitz MY Speer MD McKee et al. (2002) ArticleTitleOsteopontin inhibits mineral deposition and promotes regression of ectopic calcification Am J Pathol 161 2035–2046 Occurrence Handle1:CAS:528:DC%2BD38Xpslemur8%3D Occurrence Handle12466120 Occurrence Handle1850905

    CAS  PubMed  PubMed Central  Google Scholar 

  4. G Luo P Ducy MD McKee et al. (1997) ArticleTitleSpontaneous calcification of arteries and cartilage in mice lacking matrix GLA protein Nature 386 78–81 Occurrence Handle1:CAS:528:DyaK2sXhslCnsLk%3D Occurrence Handle9052783

    CAS  PubMed  Google Scholar 

  5. N Vyavahare M Ogle FJ Schoen et al. (1999) ArticleTitleMechanisms of bioprosthetic heart valve failure: fatigue causes collagen denaturation and glycosaminoglycan loss J Biomed Mater Res 46 44–50 Occurrence Handle1:CAS:528:DyaK1MXjt12iurk%3D Occurrence Handle10357134

    CAS  PubMed  Google Scholar 

  6. J Lovekamp N Vyavahare (2001) ArticleTitlePeriodate-mediated glycosaminoglycan stabilization in bioprosthetic heart valves J Biomed Mater Res 56 478–486 Occurrence Handle1:CAS:528:DC%2BD3MXkvFSqtLk%3D Occurrence Handle11400125

    CAS  PubMed  Google Scholar 

  7. CM Giachelli S Steitz (2000) ArticleTitleOsteopontin: a versatile regulator of inflammation and biomineralization Matrix Biol 19 615–622 Occurrence Handle1:CAS:528:DC%2BD3MXhsFClsg%3D%3D Occurrence Handle11102750

    CAS  PubMed  Google Scholar 

  8. J Sodek B Ganss MD McKee (2000) ArticleTitleOsteopontin Crit Rev Oral Biol Med 11 279–303 Occurrence Handle1:STN:280:DC%2BD3cvmsFOjtg%3D%3D Occurrence Handle11021631

    CAS  PubMed  Google Scholar 

  9. SS Srivatsa PJ Harrity PB Maercklein et al. (1997) ArticleTitleIncreased cellular expression of matrix proteins that regulate mineralization is associated with calcification of native human and porcine xenograft bioprosthetic heart valves J Clin Invest 99 996–1009 Occurrence Handle1:STN:280:DyaK2s3hs1Slug%3D%3D Occurrence Handle9062358 Occurrence Handle507908

    CAS  PubMed  PubMed Central  Google Scholar 

  10. KD O’Brien J Kuusisto DD Reichenbach et al. (1995) ArticleTitleOsteopontin is expressed in human aortic valvular lesions Circulation 92 2163–2168 Occurrence Handle7554197

    PubMed  Google Scholar 

  11. H Shiraga W Min WJ VanDusen et al. (1992) ArticleTitleInhibition of calcium oxalate crystal growth in vitro by uropontin: another member of the aspartic acid-rich protein superfamily Proc Natl Acad Sci USA 89 426–430 Occurrence Handle1:CAS:528:DyaK3sXitFSks7o%3D Occurrence Handle1729712

    CAS  PubMed  Google Scholar 

  12. AL Boskey M Maresca W Ullrich et al. (1993) ArticleTitleOsteopontin-hydroxyapatite interactions in vitro: inhibition of hydroxyapatite formation and growth in a gelatin-gel Bone Miner 22 147–159 Occurrence Handle1:CAS:528:DyaK2cXnsVagtQ%3D%3D Occurrence Handle8251766

    CAS  PubMed  Google Scholar 

  13. GK Hunter CL Kyle HA Goldberg (1994) ArticleTitleModulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin-mediated inhibition of hydroxyapatite formation Biochem J 300 723–728 Occurrence Handle1:CAS:528:DyaK2cXkt12mt78%3D Occurrence Handle8010953 Occurrence Handle1138226

    CAS  PubMed  PubMed Central  Google Scholar 

  14. T Wada MD McKee S Steitz CM Giachelli (1999) ArticleTitleCalcification of vascular smooth muscle cell cultures: inhibition by osteopontin Circ Res 84 166–178 Occurrence Handle1:CAS:528:DyaK1MXpsVGntQ%3D%3D Occurrence Handle9933248

    CAS  PubMed  Google Scholar 

  15. S Jono C Peinado CM Giachelli (2000) ArticleTitlePhosphorylation of osteopontin is required for inhibition of vascular smooth muscle cell calcification J Biol Chem 275 20197–20203 Occurrence Handle1:CAS:528:DC%2BD3cXkslags70%3D Occurrence Handle10766759

    CAS  PubMed  Google Scholar 

  16. LL Smith HK Cheung LE Ling et al. (1996) ArticleTitleOsteopontin N-terminal domain contains a cryptic adhesive sequence recognized by alpha9beta1 integrin J Biol Chem 271 IssueID45 28485–28491 Occurrence Handle1:CAS:528:DyaK28XmvVegsbo%3D Occurrence Handle8910476

    CAS  PubMed  Google Scholar 

  17. L Liaw DE Birk CB Ballas JS Whitsitt JM Davidson BL Hogan (1998) ArticleTitleAltered wound healing in mice lacking a functional osteopontin gene (spp1) J Clin Invest 101 1468–1478 Occurrence Handle1:CAS:528:DyaK1cXitleksbw%3D Occurrence Handle9525990 Occurrence Handle508725

    CAS  PubMed  PubMed Central  Google Scholar 

  18. SN Meloan H Puchtler LS Valentine (1972) ArticleTitleAlkaline and acid alizarin red S stains for alkali-soluble and alkali-insoluble calcium deposits Arch Pathol 93 190–197 Occurrence Handle1:CAS:528:DyaE38Xht1eku7o%3D Occurrence Handle4110754

    CAS  PubMed  Google Scholar 

  19. CE Murry CM Giachelli SM Schwartz R Vracko (1994) ArticleTitleMacrophages express osteopontin during repair of myocardial necrosis Am J Pathol 145 1450–1462 Occurrence Handle1:CAS:528:DyaK2MXjtV2mtb4%3D Occurrence Handle7992848 Occurrence Handle1887495

    CAS  PubMed  PubMed Central  Google Scholar 

  20. TA Horbett (1981) ArticleTitleAdsorption of proteins from plasma to a series of hydrophilic-hydrophobic copolymers. II. Compositional analysis with the prelabeled protein technique J Biomed Mater Res 15 673–695 Occurrence Handle1:CAS:528:DyaL3MXlslGkt7k%3D Occurrence Handle12659135

    CAS  PubMed  Google Scholar 

  21. PJ, TE Schryer JA Starr JTM Wright (1986) Anti-calcification effect of glutaraldehyde-preserved valve tissue stored for increasing time in glutaraldehyde EYM Bodnar (Eds) Biologic and bioprosthetic valves. Proceedings of the Third International Symposium Yorke Medical Books New York 471–477

    Google Scholar 

  22. Lawford PV, KC, Rogers S, Black MM (1993) Inhibition of calcification of porcine bioprostheses by prolonged glutaraldehyde fixation. Cardiovascular Science and Technology Conference, AAMI/NIH.

  23. G Golomb FJ Schoen MS Smith J Linden M Dixon RJ Levy (1987) ArticleTitleThe role of glutaraldehyde-induced cross-links in calcification of bovine pericardium used in cardiac valve bioprostheses Am J Pathol 127 122–130 Occurrence Handle1:CAS:528:DyaL2sXksFKkuro%3D Occurrence Handle3105321 Occurrence Handle1899585

    CAS  PubMed  PubMed Central  Google Scholar 

  24. RH Clark AA Campbell LA Klumb CJ Long PS Stayton (1999) ArticleTitleProtein electrostatic surface distribution can determine whether calcium oxalate crystal growth is promoted or inhibited Calcif Tissue Int 64 IssueID6 516–521 Occurrence Handle1:CAS:528:DyaK1MXjs1Ogsbk%3D Occurrence Handle10341024

    CAS  PubMed  Google Scholar 

  25. PA Raj M Johnsson MJ Levine GH Nancollas (1992) ArticleTitleSalivary statherin. Dependence on sequence, charge, hydrogen-bonding potency, and helical conformation for adsorption to hydroxyapatite and inhibition of mineralization J Biol Chem 267 IssueID9 5968–5976 Occurrence Handle1:CAS:528:DyaK38XhvFWmu7c%3D Occurrence Handle1313424

    CAS  PubMed  Google Scholar 

  26. JA Wesson EM Worcester JG Kleinman (2000) ArticleTitleRole of anionic proteins in kidney stone formation: interaction between model anionic polypeptides and fit calcium oxalate crystals J Urol 163 IssueID4 1343–1348 Occurrence Handle1:CAS:528:DC%2BD3cXisVSrurY%3D Occurrence Handle10737541

    CAS  PubMed  Google Scholar 

  27. HA Goldberg KJ Warner MJ Stillman GK Hunter (1996) ArticleTitleDetermination of the hydroxyapatite-nucleating region of bone sialoprotein Connect Tissue Res 35 IssueID1–4 385–392 Occurrence Handle1:CAS:528:DyaK2sXhvFals70%3D Occurrence Handle9084679

    CAS  PubMed  Google Scholar 

  28. GK Hunter CL Kyle HA Goldberg (1994) ArticleTitleModulation of crystal formation by bone phosphoproteins: structural specificity of the osteopontin- mediated inhibition of hydroxyapatite formation Biochem J 300 . IssueIDPt 3 723–728

    Google Scholar 

  29. RJ Kok AJ Schraa EJ Bos et al. (2002) ArticleTitlePreparation and functional evaluation of RGD-modified proteins as alpha(v)beta(3) integrin-directed therapeutics Bioconjug Chem 13 128–135 Occurrence Handle1:CAS:528:DC%2BD3MXos12it7c%3D Occurrence Handle11792188

    CAS  PubMed  Google Scholar 

  30. Y Katayama CM House N Udagawa et al. (1998) ArticleTitleCasein kinase 2 phosphorylation of recombinant rat osteopontin enhances adhesion of osteoclasts but not osteoblasts J Cell Physiol 176 179–187 Occurrence Handle1:CAS:528:DyaK1cXjsFGrtLg%3D Occurrence Handle9618157

    CAS  PubMed  Google Scholar 

  31. A Linde A Lussi MA Crenshaw (1989) ArticleTitleMineral induction by immobilized polyanionic proteins Calcif Tissue Int 44 286–295 Occurrence Handle1:CAS:528:DyaL1MXktVakur8%3D Occurrence Handle2501010

    CAS  PubMed  Google Scholar 

  32. A Lussi A Linde (1993) ArticleTitleMineral induction in vivo by dentine proteins Caries Res 27 241–248 Occurrence Handle1:CAS:528:DyaK2cXhtVegsrw%3D Occurrence Handle8402795

    CAS  PubMed  Google Scholar 

  33. Y Doi T Horiguchi SH Kim et al. (1993) ArticleTitleImmobilized DPP and other proteins modify OCP formation Calcif Tissue Int 52 139–145 Occurrence Handle1:CAS:528:DyaK3sXitFSjur0%3D Occurrence Handle8443690

    CAS  PubMed  Google Scholar 

  34. AL Boskey (1995) ArticleTitleOsteopontin and related phosphorylated sialoproteins: effects on mineralization Ann N Y Acad Sci 760 249–256 Occurrence Handle1:CAS:528:DyaK2MXovVSitrY%3D Occurrence Handle7785899

    CAS  PubMed  Google Scholar 

  35. FP Reinholt K Hultenby A Oldberg D Heinegard (1990) ArticleTitleOsteopontin–a possible anchor of osteoclasts to bone Proc Natl Acad Sci USA 87 4473–4475 Occurrence Handle1:CAS:528:DyaK3cXksFKhtL0%3D Occurrence Handle1693772

    CAS  PubMed  Google Scholar 

  36. ME Flores M Norgard D Heinegard FP Reinholt G Andersson (1992) ArticleTitleRGD-directed attachment of isolated rat osteoclasts to osteopontin, bone sialoprotein, and fibronectin Exp Cell Res 201 526–530 Occurrence Handle1:CAS:528:DyaK38XlsVyjt70%3D Occurrence Handle1639145

    CAS  PubMed  Google Scholar 

  37. FP Ross J Chappel JI Alvarez et al. (1993) ArticleTitleInteractions between the bone matrix proteins osteopontin and bone sialoprotein and the osteoclast integrin alpha v beta 3 potentiate bone resorption J Biol Chem 268 9901–9907 Occurrence Handle1:CAS:528:DyaK3sXktVGrtb4%3D Occurrence Handle8486670

    CAS  PubMed  Google Scholar 

  38. M Sato MK Sardana WA Grasser VM Garsky JM Murray RJ Gould (1990) ArticleTitleEchistatin is a potent inhibitor of bone resorption in culture J Cell Biol 111 1713–1723 Occurrence Handle1:CAS:528:DyaK3cXlvVKgtbY%3D Occurrence Handle2211834

    CAS  PubMed  Google Scholar 

  39. MA Horton ML Taylor TR Arnett MH Helfrich (1991) ArticleTitleArg-Gly-Asp (RGD) peptides and the anti-vitronectin receptor antibody 23C6 inhibit dentine resorption and cell spreading by osteoclasts Exp Cell Res 195 368–375 Occurrence Handle1:CAS:528:DyaK38XhvVymsw%3D%3D Occurrence Handle1712731

    CAS  PubMed  Google Scholar 

  40. JE Fisher MP Caulfield M Sato et al. (1993) ArticleTitleInhibition of osteoclastic bone resorption in vivo by echistatin, an “arginyl-glycyl-aspartyl” (RGD)-containing protein Endocrinology 132 1411–1413 Occurrence Handle1:CAS:528:DyaK3sXksVSrtrg%3D Occurrence Handle8440195

    CAS  PubMed  Google Scholar 

  41. MW Lark GB Stroup RA Dodds et al. (2001) ArticleTitleAntagonism of the osteoclast vitronectin receptor with an orally active nonpeptide inhibitor prevents cancellous bone loss in the ovariectomized rat J Bone Miner Res 16 319–327 Occurrence Handle1:CAS:528:DC%2BD3MXptlOqtA%3D%3D Occurrence Handle11204432

    CAS  PubMed  Google Scholar 

  42. H Yoshitake SR Rittling DT Denhardt M Noda (1999) ArticleTitleOsteopontin-deficient mice are resistant to ovariectomy-induced bone resorption Proc Natl Acad USA 96 8156–8160 Occurrence Handle1:CAS:528:DyaK1MXltVOrsL8%3D

    CAS  Google Scholar 

  43. Y Asou SR Rittling H Yoshitake et al. (2001) ArticleTitleOsteopontin facilitates angiogenesis, accumulation of osteoclasts, and resorption in ectopic bone Endocrinology 142 1325–1332 Occurrence Handle1:CAS:528:DC%2BD3MXhsFemtbg%3D Occurrence Handle11181551

    CAS  PubMed  Google Scholar 

  44. N Tani-Ishii A Tsunoda T Umemoto (1997) ArticleTitleOsteopontin antisense deoxyoligonucleotides inhibit bone resorption by mouse osteoclasts in vitro J Periodontal Res 32 480–645 Occurrence Handle1:CAS:528:DyaK2sXmsVanu7c%3D Occurrence Handle9379315

    CAS  PubMed  Google Scholar 

  45. B Ek-Rylander M Flores M Wendel D Heinegard G Andersson (1994) ArticleTitleDephosphorylation of osteopontin and bone sialoprotein by osteoclastic tartrate-resistant acid phosphatase. Modulation of osteoclast adhesion in vitro J Biol Chem 269 14853–14856 Occurrence Handle1:CAS:528:DyaK2cXktVGrsrw%3D Occurrence Handle8195113

    CAS  PubMed  Google Scholar 

  46. S Razzouk JC Brunn C Qin CE Tye HA Goldberg WT Butler (2002) ArticleTitleOsteopontin posttranslational modifications, possibly phosphorylation, are required for in vitro bone resorption but not osteoclast adhesion Bone 30 40–47 Occurrence Handle1:CAS:528:DC%2BD38XktFeqsA%3D%3D Occurrence Handle11792563

    CAS  PubMed  Google Scholar 

  47. AV Rousselle D Heymann (2002) ArticleTitleOsteoclastic acidification pathways during bone resorption Bone 30 533–540 Occurrence Handle1:CAS:528:DC%2BD38XisV2ksL8%3D Occurrence Handle11934642

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

Studies were supported by NSF grant EEC-9529161 and NIH grant HL62329. The authors thank Edwards Lifesciences Inc. for their gift of glutaraldehyde-fixed bovine pericardium, and Kelly Simon and Abhi Banerjee for their technical help with histological staining and image analysis. The authors acknowledge use of the University of Washington Engineered Biomaterials (UWEB) Optical Microscopy and Image Analysis Shared Resource, funded by the National Science Foundation through grants EEC-9872882 and EEC-9529161.

Author information

Authors and Affiliations

  1. Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA

    Rachit Ohri, Elyse Tung, Rupak Rajachar & Cecilia M. Giachelli

Authors
  1. Rachit Ohri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elyse Tung
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rupak Rajachar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cecilia M. Giachelli
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cecilia M. Giachelli.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ohri, R., Tung, E., Rajachar, R. et al. Mitigation of Ectopic Calcification in Osteopontin-Deficient Mice by Exogenous Osteopontin. Calcif Tissue Int 76, 307–315 (2005). https://doi.org/10.1007/s00223-004-0071-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00223-004-0071-7

Keywords

Search

Navigation