Journal of Bionic Engineering

, Volume 12, Issue 4, pp 604–612 | Cite as

Enhanced Osteogenesis of Nanosized Cobalt-substituted Hydroxyapatite

  • Nenad Ignjatović
  • Zorica Ajduković
  • Jelena Rajković
  • Stevo Najman
  • Dragan Mihailović
  • Dragan UskokovićEmail author


Hydroxyapatite (HAp) is an extensively studied material with known biocompatible and osteoconductive properties in bone tissue reconstruction. The improvement of the osteogenetic potential of HAp has been tested through modification of its structure, by replacing Ca2+ ions with Co2+ ions. In our study, we comparatively analyze the osteogenetic potential of the synthesized HAp and Co2+-substituted HAp (HAp/Co) designed on the nano-scale with the aim of specifically stimulating osteogenesis in vivo. We present a quantitative study of the microscopic organization and structure of the newly formed tissue in a bone defect after 12 weeks and 24 weeks. A quantitative analysis of the calcium, magnesium and phosphorus content in the defect and its close environment was used to determine the deposition of minerals after bone reconstruction. The defect reconstructed with HAp/Co nanoparticles (Co2+ content 12 wt%) was filled with a new tissue matrix composed of dense collagen fibers containing centers of mineralization after 24 weeks. The mineral deposition rate was also higher when the defect was reconstructed with HAp/Co than when it was filled with pure HAp. A histological analysis confirmed that the alveolar bone, in which osteoporosis-induced defects were repaired using HAp/Co nanoparticles, was recuperated.


advanced osteogenesis cobalt-substituted hydroxyapatite nanoparticles histomorphometry 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    Eriksen E, Keaveny T, Gallagher E, Krege J. Literature review: The effects of teriparatide therapy at the hip in patients with osteoporosis. Bone, 2014, 67, 246–256.CrossRefGoogle Scholar
  2. [2]
    Samavedi S, Whittington A, Goldstein A. Calcium phosphate ceramics in bone tissue engineering: A review of properties and their influence on cell behavior. Acta Bio-materialia, 2013, 9, 8037–8045.CrossRefGoogle Scholar
  3. [3]
    Parsons A J, Ahmed I, Han N, Felfel R, Rudd C D. Mimicking bone structure and function with structural composite materials. Journal of Bionic Engineering, 2010, 7, S1–S10.CrossRefGoogle Scholar
  4. [4]
    Dorozhkin S. Calcium orthophosphates in nature, biology and medicine. Materials, 2009, 2, 399–498.CrossRefGoogle Scholar
  5. [5]
    Lin K, Wu C, Chang J. Advances in synthesis of calcium phosphate crystals with controlled size and shape. Acta Biomaterialia, 2014, 10, 4071–4102.CrossRefGoogle Scholar
  6. [6]
    Uskoković V, Uskoković D. Nanosized hydroxyapatite and other calcium phosphates: Chemistry of formation and application as drug and gene delivery agents. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2011, 96, 152–191.CrossRefGoogle Scholar
  7. [7]
    Dorozhkin S. Nanosized and nanocrystalline calcium orthophosphates. Acta Biomaterialia, 2010, 6, 715–734.CrossRefGoogle Scholar
  8. [8]
    Thian E, Konishi T, Kawanobe Y, Lim P, Choong C, Ho B, Aizawa M. Zinc-substituted hydroxyapatite: A biomaterial with enhanced bioactivity and antibacterial properties. Journal of Materials Science: Materials in Medicine, 2013, 24, 437–445.Google Scholar
  9. [9]
    Cox S, Jamshidi P, Grover L, Mallick K. Preparation and characterisation of nanophase Sr, Mg, and Zn substituted hydroxyapatite by aqueous precipitation. Materials Science and Engineering: C, 2014, 35, 106–114.CrossRefGoogle Scholar
  10. [10]
    Stojanović Z, Veselinović Lj, Marković S, Ignjatović N. Uskoković D. Hydrothermal synthesis of nanosize pure and cobalt-exchanged hydroxyapatite. Materials and Manufacturing Processes, 2009, 24, 1096–1103.CrossRefGoogle Scholar
  11. [11]
    Kramer E, Itzkowitz E, Wei M. Synthesis and characterization of cobalt-substituted hydroxyapatite powders. Ceramics International, 2014, 40, 13471–13480.CrossRefGoogle Scholar
  12. [12]
    Tank K, Chudasama K, Thaker V, Joshi M. Cobalt-doped nanohydroxyapatite: Synthesis, characterization, antimicrobial and hemolytic studies. Journal of Nanoparticle Research, 2013, 15, 1644–1655.CrossRefGoogle Scholar
  13. [13]
    Darolles C, Sage N, Armengaud J, Malard V. In vitro assessment of cobalt oxide particle toxicity: Identifying and circumventing interference. Toxicology in Vitro, 2013, 27, 1699–1710.CrossRefGoogle Scholar
  14. [14]
    Smith L, Holmes A, Kumar Kandpal S, Mason M, Zheng T, Wise J Sr. The cytotoxicity and genotoxicity of soluble and particulate cobalt in human lung fibroblast cells. Toxicology and Applied Pharmacology, 2014, 278, 259–265.CrossRefGoogle Scholar
  15. [15]
    Fan W, Crawford R, Xiao Y. Enhancing in vivo vascularized bone formation by cobalt chloride-treated bone marrow stromal cells in a tissue engineered periosteum model. Biomaterials, 2010, 31, 3580–3589.CrossRefGoogle Scholar
  16. [16]
    Bose S, Fielding G, Tarafder S, Bandyopadhyay A. Understanding of dopant-induced osteogenesis and angiogenesis in calcium phosphate ceramics. Trends in Biotechnology, 2013, 31, 594–605.CrossRefGoogle Scholar
  17. [17]
    Wu C, Zhou Y, Fan W, Han P, Chang J, Yuen J, Zhang M, Xiao Y. Hypoxia-mimicking mesoporous bioactive glass scaffolds with controllable cobalt ion release for bone tissue engineering. Biomaterials, 2012, 33, 2076–2085.CrossRefGoogle Scholar
  18. [18]
    Veselinović Lj, Karanović Lj, Stojanović Z, Bracko I, Marković S, Ignjatović N, Uskoković D. Crystal structure of cobalt-substituted calcium hydroxyapatite nanopowders prepared by hydrothermal processing. Journal of Applied Crystallography, 2010, 43, 320–327.CrossRefGoogle Scholar
  19. [19]
    Ignjatović N, Ajduković Z, Savić V, Najman S, Mihailović D, Vasiljević P, Stojanović Z, Uskoković V, Uskoković D. Nanoparticles of cobalt-substituted hydroxyapatite in regeneration of mandibular osteoporotic bones. Journal of Materials Science: Materials in Medicine, 2013, 24, 343–354.Google Scholar
  20. [20]
    Ajduković Z, Najman S, Djordjević Lj, Savić V, Petrović D, Ignjatović N, Uskoković D. Repair of bone tissue affected by osteoporosis with hydroxyapatite-poly-L-(HAp/PLLA) with and without blood plasma. Journal of Biomaterials Application, 2005, 20, 179–190.CrossRefGoogle Scholar
  21. [21]
    Dempster D, Compston J, Drezner M, Glorieux F, Kanis J, Malluche H, Meunier P, Ott S, Recker R, Parfitt M. Standardized nomenclature, symbols, and units for bone histo-morphometry: A 2012 update of the report of the ASBMR histomorphometry nomenclature committee. Journal of Bone and Mineral Research, 2013, 28, 2–17.CrossRefGoogle Scholar
  22. [22]
    Drašler B, Drobne D, Novak S, Valant J, Boljte S, Otrin L, Rappolt M, Sartori B, Iglič A, Kralj-Iglič V, Šuštar V. Makovec D, Gyergyek S, Hočevar M, Godec M, Zupanc J. Effects of magnetic cobalt ferrite nanoparticles on biological and artificial lipid membranes. International Journal of Nanomedicine, 2014, 9, 1559–1581.CrossRefGoogle Scholar
  23. [23]
    Simonsen L, Harbak H, Bennekou P. Cobalt metabolism and toxicology–a brief update. Science of the Total Environment, 2012, 432, 210–215.CrossRefGoogle Scholar
  24. [24]
    Maier J, Bernardini D, Rayssiguier Y, Mazur A. High concentrations of magnesium modulate vascular endothelial cell behaviour in vitro. Biochimica et Biophysica Acta (BBA) -Molecular Basis of Disease, 2004, 1689, 6–12.CrossRefGoogle Scholar
  25. [25]
    Cooke J, Losordo D. Nitric oxide and angiogenesis. Circulation, 2002, 105, 2133–2135.CrossRefGoogle Scholar
  26. [26]
    Ripamonti U. Biomimetism, biomimetic matrices and the induction of bone formation. Journal of Cellular and Molecular Medicine, 2009, 13, 2953–2972.CrossRefGoogle Scholar
  27. [27]
    Nudelman F, Pieterse K, George A, Bomans P, Friedrich H, Brylka L, Hilbers P, de With G, Sommerdijk N. The role of collagen in bone apatite formation in the presence of hydroxyapatite nucleation inhibitors. Nature Materials, 2010, 9, 1004–1009.CrossRefGoogle Scholar
  28. [28]
    Mata A, Geng Y, Henrikson J, Aparicio C, Stock R, Satcher R, Stupp I. Bone regeneration mediated by biomimetic mineralization of a nanofiber matrix. Biomaterials, 2010, 31, 6004–6012.CrossRefGoogle Scholar
  29. [29]
    Kikuchi M, Ikoma T, Itoh S, Matsumoto N, Koyama Y, Takakuda K, Shinomiya K, Tanaka J. Biomimetic synthesis of bone-like nanocomposites using the self-organization mechanism of hydroxyapatite and collagen. Composites Science and Technology, 2004, 64, 819–825.CrossRefGoogle Scholar
  30. [30]
    Ryadnov M, Woolfson D. Engineering the morphology of a self-assembling protein fibre. Nature Materials, 2003, 2, 329–332.CrossRefGoogle Scholar

Copyright information

© Jilin University 2015

Authors and Affiliations

  • Nenad Ignjatović
    • 1
  • Zorica Ajduković
    • 2
  • Jelena Rajković
    • 3
  • Stevo Najman
    • 3
  • Dragan Mihailović
    • 4
  • Dragan Uskoković
    • 1
    Email author
  1. 1.Centre for Fine Particles Processing and NanotechnologiesInstitute of Technical Sciences of the Serbian Academy of Science and ArtsBelgradeSerbia
  2. 2.Faculty of Medicine, Department of Prosthodontics, Clinic of StomatologyUniversity of NišNišSerbia
  3. 3.Department of Biology and Ecology Faculty of Science and MathematicsUniversity of NišNišSerbia
  4. 4.Faculty of Medicine, Institute of PathologyUniversity of NišNišSerbia

Personalised recommendations