Calcified Tissue International

, Volume 76, Issue 2, pp 146–153 | Cite as

Localization of Pigment Epithelium-Derived Factor in Growing Mouse Bone

  • Gerald M. Y. Quan
  • Joseline Ojaimi
  • Yaping Li
  • Vicky Kartsogiannis
  • Hong Zhou
  • Peter F. M. Choong


Pigment epithelium-derived factor (PEDF) is a potent anti-angiogenic factor found in a wide range of fetal and adult tissues, where it is thought to play a role in the regulation of angiogenesis during development. The temporal expression of PEDF during endochondral bone formation has not previously been reported. In this study, we analysed the expression pattern of PEDF in growing mouse hindlimbs from newborn day one through to maturation at week 9, using immunohistochemistry and in situ hybridization. PEDF expression was demonstrated in chondrocytes within the resting, proliferative and upper hypertrophic zones of the epiphyseal growth plate. The pattern of expression was consistent throughout the developmental stages of the mouse. In addition, PEDF was expressed by osteoblasts lining the bone spicules in the ossification zone of metaphyseal bone, as well as by osteoblasts lining cortical periosteum. These novel results demonstrate that PEDF is developmentally expressed in both cartilage and bone cells during endochondral bone formation, and strongly suggest that it may play a regulatory role in the processes of chondrocyte and osteoblast differentiation, endochondral ossification, and bone remodelling during growth and development of long bones.


PEDF Chondrocyte Osteoblast Growth plate Cartilage Endochondral ossification Bone 



This study was supported by the Victorian Orthopaedic Research Trust, Australian Orthopaedic Association, and the Anti-Cancer Council of Victoria. The first author is a recipient of scholarships awarded by the Faculty of Medicine, University of Melbourne, and the National Health and Medical Research Council of Australia. The authors would like to thank Professor T. J. Martin for his helpful advice regarding the manuscript.


  1. 1.
    Tombran-Tink, J, Chader, GG, Johnson, LV 1991PEDF: a pigment epithelium-derived factor with potent neuronal differentiative activityExp Eye Res53411414CrossRefPubMedGoogle Scholar
  2. 2.
    Steele, FR, Chader, GJ, Johnson, LV, Tombran-Tink, J 1993Pigment epithelium-derived factor: neurotrophic activity and identification as a member of the serine protease inhibitor gene familyProc Natl Acad Sci USA9015261530PubMedGoogle Scholar
  3. 3.
    Araki, T, Taniwaki, T, Becerra, SP, Chader, GJ, Schwartz, JP 1998Pigment epithelium-derived factor (PEDF) differentially protects immature but not mature cerebellar granule cells against apoptotic cell deathJ Neurosci Res53715CrossRefPubMedGoogle Scholar
  4. 4.
    Houenou, LJ, D’Costa, AP, Li, L, Turgeon, VL, Enyadike, C, Alberdi, E, Becerra, SP 1999Pigment epithelium-derived factor promotes the survival and differentiation of developing spinal motor neuronsJ Comp Neurol412506514CrossRefPubMedGoogle Scholar
  5. 5.
    Pignolo, RJ, Cristofalo, VJ, Rotenberg, MO 1993Senescent WI-38 cells fail to express EPC-1, a gene induced in young cells upon entry into the G0 stateJ Biol Chem26889498957PubMedGoogle Scholar
  6. 6.
    Dawson, DW, Volpert, OV, Gillis, P, Crawford, SE, Xu, H, Benedict, W, Bouck, NP 1999Pigment epithelium-derived factor: a potent inhibitor of angiogenesisScience285245248CrossRefPubMedGoogle Scholar
  7. 7.
    Crawford, SE, Stellmach, V, Ranalli, M,  et al. 2001Pigment epithelium-derived factor (PEDF) in neuroblastoma: a multifunctional mediator of Schwann cell antitumor activityJ Cell Sci11444214428PubMedGoogle Scholar
  8. 8.
    Doll, JA, Stellmach, VM, Bouck, NP,  et al. 2003Pigment epithelium-derived factor regulates the vasculature and mass of the prostate and pancreasNat Med9774780CrossRefPubMedGoogle Scholar
  9. 9.
    Abramson, LP, Stellmach, V, Doll, JA, Cornwell, M, Arensman, RM, Crawford, SE 2003Wilms’ tumor growth is suppressed by antiangiogenic pigment epithelium-derived factor in a xenograft modelJ Pediatr Surg38336342CrossRefPubMedGoogle Scholar
  10. 10.
    Meyer, C, Notari, L, Becerra, SP 2002Mapping the type I collagen-binding site on pigment epithelium-derived factorImplications for its antiangiogenic activity. J Biol Chem2774540045407Google Scholar
  11. 11.
    Tombran-Tink, J, Mazuruk, K, Rodriguez, IR, Chung, D, Linker, T, Englander, E, Chader, GJ 1996Organization, evolutionary conservation, expression and unusual Alu density of the human gene for pigment epithelium-derived factor, a unique neurotrophic serpinMol Vis2111Google Scholar
  12. 12.
    Wu, YQ, Becerra, SP 1996Proteolytic activity directed toward pigment epithelium-derived factor in vitreous of bovine eyesImplications of proteolytic processing. Invest Ophthalmol Vis Sci3719841993Google Scholar
  13. 13.
    Gao, G, Li, Y, Zhang, D, Gee, S, Crosson, C, Ma, J 2001Unbalanced expression of VEGF and PEDF in ischemia-induced retinal neovascularizationFEES Lett489270276CrossRefGoogle Scholar
  14. 14.
    Ogata, N, Nishikawa, M, Nishimura, T, Mitsuma, Y, Matsumura, M 2002Inverse levels of pigment epithelium-derived factor and vascular endothelial growth factor in the vitreous of eyes with rhegmatogenous retinal detachment and proliferative vitreoretinopathyAm J Ophthalmol133851852CrossRefPubMedGoogle Scholar
  15. 15.
    Horner, A, Bishop, NJ, Bord, S, Beeton, C, Kelsall, AW, Coleman, N, Compston, JE 1999Immunolocalisation of vascular endothelial growth factor (VEGF) in human neonatal growth plate cartilageJ Anat194519524CrossRefPubMedGoogle Scholar
  16. 16.
    Jingushi, S, Scully, SP, Joyce, ME, Sugioka, Y, Bolander, ME 1995Transforming growth factor-beta 1 and fibroblast growth factors in rat growth plateJ Orthop Res13761768CrossRefPubMedGoogle Scholar
  17. 17.
    Carlevaro, MF, Albini, A, Ribatti, D,  et al. 1997Transferrin promotes endothelial cell migration and invasion: implication in cartilage neovascularizationJ Cell Biol13613751384CrossRefPubMedGoogle Scholar
  18. 18.
    Shukunami, C, Iyama, K, Inoue, H, Hiraki, Y 1999Spatiotemporal pattern of the mouse chondromodulin-I gene expression and its regulatory role in vascular invasion into cartilage during endochondral bone formationInt J Dev Biol433949PubMedGoogle Scholar
  19. 19.
    Cheung, WH, Lee, KM, Fung, KP, Leung, KS 2001Growth plate chondrocytes inhibit neo-angiogenesis — a possible mechanism for tumor controlCancer Lett1632532CrossRefPubMedGoogle Scholar
  20. 20.
    Quan, GMY, Ojaimi, J, Nadesapillai, APW, Zhou, H, Choong, PFM 2002Resistance of epiphyseal cartilage to invasion by osteosarcoma is likely to be due to the expression of anti-angiogenic factorsPathobiology70361367CrossRefPubMedGoogle Scholar
  21. 21.
    Fisher, JL, Field, CL, Zhou, H, Harris, TL, Henderson, MA, Choong, PFM 2000Urokinase plasminogen activator system gene expression is increased in human breast carcinoma and its bone metastases—a comparison of normal breast tissue, non-invasive and invasive carcinoma and osseous metastasesBreast Cancer Res Treat61112CrossRefPubMedGoogle Scholar
  22. 22.
    Hiraki, Y, Inoue, H, Iyama, K,  et al. 1997Identification of chondromodulin I as a novel endothelial cell growth inhibitorPurification and its localization in the avascular zone of epiphyseal cartilage, J Biol Chem2723241932426Google Scholar
  23. 23.
    Moses, MA, Wiederschain, D, Wu, I,  et al. 1999Troponin I is present in human cartilage and inhibits angiogenesisProc Natl Acad Sci USA9626452650CrossRefPubMedGoogle Scholar
  24. 24.
    Liu, N, Lapcevich, RK, Underhill, CB,  et al. 2001Metastatin: a hyaluronan-binding complex from cartilage that inhibits tumor growthCancer Res6110221028PubMedGoogle Scholar
  25. 25.
    Cheung, WH, Lee, KM, Fung, KP, Lui, PY, Leung, KS 2001TGF-beta 1 is the factor secreted by proliferative chondrocytes to inhibit neo-angiogenesisJ Cell Biochem (suppl)367988Google Scholar
  26. 26.
    Garcia-Ramirez, M, Toran, N, Andaluz, P, Carrascosa, A, Audi, L 2000Vascular endothelial growth factor is expressed in human fetal growth cartilageJ Bone Miner Res15534540PubMedGoogle Scholar
  27. 27.
    Carlevaro, MF, Cermelli, S, Cancedda, R, Descalzi Cancedda, F 2000Vascular endothelial growth factor (VEGF) in cartilage neovascularization and chondrocyte differentiation: auto-paracrine role during endochondral bone formationJ Cell Sci1135969PubMedGoogle Scholar
  28. 28.
    Alini, M, Marriott, A, Chen, T, Abe, S, Poole, AR 1996A novel angiogenic molecule produced at the time of chondrocyte hypertrophy during endochondral bone formationDev Biol176124132CrossRefPubMedGoogle Scholar
  29. 29.
    Gerber, HP 1999Vascular endothelial growth factor plays an important part in longitudinal bone growth by stimulating angiognesis into the growth plateNat Med5623628CrossRefPubMedGoogle Scholar
  30. 30.
    Yin, M, Gentili, C, Koyama, E, Zasloff, M, Pacifici, M 2002Antiangiogenic treatment delays chondrocyte maturation and bone formation during limb skeletogenesisJ Bone Miner Res175665PubMedGoogle Scholar
  31. 31.
    Shao, H, Schvartz, I, Shaltiel, S 2003Secretion of pigment epithelium-derived factorEur J Biochem270822831CrossRefPubMedGoogle Scholar
  32. 32.
    Petersen, SV, Valnickova, Z, Enghild, JJ 2003A physiologically relevant concentration of pigment epithelium-derived factor (PEDF) in human bloodPurification and characterization. Biochem J374199206Google Scholar
  33. 33.
    Simonovic, M, Gettins, PG, Volz, K 2001Crystal structure of human PEDF, a potent anti-angiogenic and neurite growth-promoting factorProc Natl Acad Sci USA981113111135CrossRefPubMedGoogle Scholar
  34. 34.
    Yasui, N, Mori, T, Morito, D, Matsushita, O, Kourai, H, Nagata, K, Koide, T 2003Dual-site recognition of different extracellular matrix components by anti-angiogenic/neurotrophic serpin, PEDFBiochemistry4231603167CrossRefPubMedGoogle Scholar
  35. 35.
    Kozaki, K, Miyaishi, O, Koiwai, O,  et al. 1998Isolation, purification, and characterization of a collagen-associated serpin, caspin, produced by murine colon adenocarcinoma cellsJ Biol Chem2731512515130CrossRefPubMedGoogle Scholar
  36. 36.
    Tombran-Tink, J, Barnstable, CJ 2004Osteoblasts and osteoclasts express PEDF, VEGF-A isoforms and VEGF receptors: possible mediators of angiogenesis and matrix remodeling in the boneBiochem Biophys Res Commun316573579CrossRefPubMedGoogle Scholar
  37. 37.
    Stellmach, V, Crawford, SE, Zhou, W, Bouck, N 2001Prevention of ischemia-induced retinopathy by the natural ocular antiangiogenic agent pigment epithelium-derived factorProc Natl Acad Sci USA9825932597CrossRefPubMedGoogle Scholar
  38. 38.
    Volpert, OV, Zaichuk, T, Zhou, W, Reiher, F, Ferguson, TA, Stuart, PM, Amin, M, Bouck, NP 2002Inducer-stimulated Fas targets activated endothelium for destruction by anti-angiogenic thrombospondin-1 and pigment epithelium-derived factorNat Med8349357CrossRefPubMedGoogle Scholar
  39. 39.
    Aizawa, T, Roach, HI, Kokubun, S, Tanaka, Y 1998Changes in the expression of fas, osteonectin and osteocalcin with age in the rabbit growth plateJ Bone Joint Surg Br80880887CrossRefPubMedGoogle Scholar
  40. 40.
    Hashimoto, S, Setareh, M, Ochs, RL, Lotz, M 1997Fas/Fas ligand expression and induction of apoptosis in chondrocytesArthritis Rheum4017491755PubMedGoogle Scholar
  41. 41.
    Tombran-Tink, J, Pawar, H, Swaroop, A, Rodriguez, I, Chader, GJ 1994Localization of the gene for pigment epithelium-derived factor (PEDF) to chromosome 17p13.1 and expression in cultured human retinoblastoma cellsGenomics19266272CrossRefPubMedGoogle Scholar
  42. 42.
    Seigel, GM, Tombran-Tink, J, Becerra, SP, Chader, GJ, Diloreto, DA,Jr., Cerro, C, Lazar, ES, Cerro, M 1994Differentiation of Y79 retinoblastoma cells with pigment epithelial- derived factor and interphotoreceptor matrix wash: effects on tumorigenicityGrowth Factors10289297PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2004

Authors and Affiliations

  • Gerald M. Y. Quan
    • 1
  • Joseline Ojaimi
    • 1
  • Yaping Li
    • 1
  • Vicky Kartsogiannis
    • 2
  • Hong Zhou
    • 2
  • Peter F. M. Choong
    • 1
    • 3
  1. 1.Department of OrthopaedicsUniversity of Melbourne, St. Vincent’s Hospital MelbourneFitzroyAustralia
  2. 2.Department of MedicineUniversity of Melbourne, St. Vincent’s Hospital MelbourneFitzroyAustralia
  3. 3.Division of Surgical OncologyPeter MacCallum Cancer InstituteEast MelbourneAustralia

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