Applied Biochemistry and Biotechnology

, Volume 186, Issue 3, pp 779–788 | Cite as

Valorization of Bone Waste of Saudi Arabia by Synthesizing Hydroxyapatite

  • Touseef AmnaEmail author


At present, hydroxyapatite is being frequently used for diverse biomedical applications as it possesses excellent biocompatibility, osteoconductivity, and non-immunogenic characteristics. The aim of the present work was to recycle bone waste for synthesis of hydroxyapatite nanoparticles to be used as bone extracellular matrix. For this reason, we for the first time utilized bio-waste of cow bones of Albaha city. The residual bones were utilized for the extraction of natural bone precursor hydroxyapatite. A facile scientific technique has been used to synthesize hydroxyapatite nanoparticles through calcinations of wasted cow bones without further supplementation of chemicals/compounds. The obtained hydroxyapatite powder was ascertained using physicochemical techniques such as XRD, SEM, FTIR, and EDX. These analyses clearly show that hydroxyapatite from native cow bone wastes is biologically and physicochemically comparable to standard hydroxyapatite, commonly used for biomedical functions. The cell viability and proliferation over the prepared hydroxyapatite was confirmed with CCk-8 colorimetric assay. The morphology of the cells growing over the nano-hydroxyapatite shows that natural hydroxyapatite promotes cellular attachment and proliferation. Hence, the as-prepared nano-hydroxyapatite can be considered as cost-effective source of bone precursor hydroxyapatite for bone tissue engineering. Taking into account the projected demand for reliable bone implants, the present research work suggested using environment friendly methods to convert waste of Albaha city into nano-hydroxyapatite scaffolds. Therefore, besides being an initial step towards accomplishment of projected demands of bone implants in Saudi Arabia, our study will also help in reducing the environmental burden by recycling of bone wastes of Albaha city.


Implants Hydroxyapatite Native cow Recycling Bones 


Funding Information

This research (Proposal No. 54-1436) was supported by the Deanship for Scientific Research, University of Albaha, Albaha, Kingdom of Saudi Arabia (KSA), funded by the Ministry of Higher Education. Prof. Dr. Touseef Amna sincerely acknowledges the research grant.

Compliance with Ethical Standards

Conflict of Interest

The author declares that he has no conflict of interest.

Ethical Statement

In the present study, there is no use of experimental animals. All the experiments were done under in vitro conditions following standard scientific procedures and ethics.


  1. 1.
    Wu, S., Liu, X., Yeung, K. W. K., Liu, C., & Yang, X. (2014). Biomimetic porous scaffolds for bone tissue engineering. Materials Science and Engineering: R: Reports, 80, 1–36.CrossRefGoogle Scholar
  2. 2.
    Cai, Y., & Tang, R. (2008). Calcium phosphate nanoparticles in biomineralization and biomaterials. Journal of Materials Chemistry, 18(32), 3775–3787.CrossRefGoogle Scholar
  3. 3.
    Tampieri, A., Sprio, S., Ruffini, A., Celotti, G., Lesci, I. G., & Roveri, N. (2009). From wood to bone: multi-step process to convert wood hierarchical structures into biomimetic hydroxyapatite scaffolds for bone tissue engineering. Journal of Materials Chemistry, 19(28), 4973–4980.CrossRefGoogle Scholar
  4. 4.
    Zhang, C., Yang, J., Quan, Z., Yang, P., Li, C., Hou, Z., & Lin, J. (2009). Hydroxyapatite nano-and microcrystals with multiform morphologies: controllable synthesis and luminescence properties. Crystal Growth and Design, 9(6), 2725–2733.CrossRefGoogle Scholar
  5. 5.
    Hou, Z., Yang, P., Lian, H., Wang, L., Zhang, C., Li, C., Chai, R., Cheng, Z., & Lin, J. (2009). Multifunctional hydroxyapatite nanofibers and microbelts as drug carriers. Chemistry–A European Journal, 15(28), 6973–6982.CrossRefGoogle Scholar
  6. 6.
    Ma, M.-Y., Zhu, Y.-J., Li, L., & Cao, S.-W. (2008). Nanostructured porous hollow ellipsoidal capsules of hydroxyapatite and calcium silicate: preparation and application in drug delivery. Journal of Materials Chemistry, 18(23), 2722–2727.CrossRefGoogle Scholar
  7. 7.
    White, A. A., Best, S. M., & Kinloch, I. A. (2007). Hydroxyapatite–carbon nanotube composites for biomedical applications: a review. International Journal of Applied Ceramic Technology, 4(1), 1–13.CrossRefGoogle Scholar
  8. 8.
    Du, C., & Wang, Y.-J. (2009). Progress in the biomineralization study of bone and enamel and biomimetic synthesis of calcium phosphate. Journal of Inorganic Materials-Beijing, 24(5), 882–888.CrossRefGoogle Scholar
  9. 9.
    Palmer, L. C., Newcomb, C. J., Kaltz, S. R., Spoerke, E. D., & Stupp, S. I. (2008). Biomimetic systems for hydroxyapatite mineralization inspired by bone and enamel. Chemical Reviews, 108(11), 4754–4783.CrossRefGoogle Scholar
  10. 10.
    Xiao, J., Zhu, Y., Ruan, Q., Liu, Y., Zeng, Y., Xu, F., & Zhang, L. (2010). Biomacromolecule and surfactant complex matrix for oriented stack of 2-dimensional carbonated hydroxyapatite nanosheets as alignment in calcified tissues. Crystal Growth & Design, 10(4), 1492–1499.CrossRefGoogle Scholar
  11. 11.
    Cheng, X., Huang, Z., Li, J., Liu, Y., Chen, C., Chi, R.-A., & Hu, Y. (2010). Self-assembled growth and pore size control of the bubble-template porous carbonated hydroxyapatite microsphere. Crystal Growth & Design, 10(3), 1180–1188.CrossRefGoogle Scholar
  12. 12.
    Cheng, X., He, Q., Li, J., Huang, Z., & Chi, R.-A. (2009). Control of pore size of the bubble-template porous carbonated hydroxyapatite microsphere by adjustable pressure. Crystal Growth and Design, 9(6), 2770–2775.CrossRefGoogle Scholar
  13. 13.
    Zhang, C., Cheng, Z., Yang, P., Xu, Z., Peng, C., Li, G., & Lin, J. (2009). Architectures of strontium hydroxyapatite microspheres: solvothermal synthesis and luminescence properties. Langmuir, 25(23), 13591–13598.CrossRefGoogle Scholar
  14. 14.
    Tan, S.-H., Chen, X.-G., Ye, Y., Sun, J., Dai, L.-Q., & Ding, Q. (2010). Hydrothermal removal of Sr 2+ in aqueous solution via formation of Sr-substituted hydroxyapatite. Journal of Hazardous Materials, 179(1-3), 559–563.CrossRefGoogle Scholar
  15. 15.
    Hui, J., Xiang, G., Xu, X., Zhuang, J., & Wang, X. (2009). Monodisperse F-substituted hydroxyapatite single-crystal nanotubes with amphiphilic surface properties. Inorganic Chemistry, 48(13), 5614–5616.CrossRefGoogle Scholar
  16. 16.
    Liu, J.-K., Cao, T.-J., Lu, Y., & Luo, C.-X. (2009). Facile preparation of assembly hydroxyapatite spheres to produce nanocomposite. Materials Technology, 24(2), 88–91.CrossRefGoogle Scholar
  17. 17.
    Shum, H. C., Bandyopadhyay, A., Bose, S., & Weitz, D. A. (2009). Double emulsion droplets as microreactors for synthesis of mesoporous hydroxyapatite. Chemistry of Materials, 21(22), 5548–5555.CrossRefGoogle Scholar
  18. 18.
    Neira, I. S., Kolen’ko, Y. V., Lebedev, O. I., Van Tendeloo, G., Gupta, H. S., Guitián, F., & Yoshimura, M. (2008). An effective morphology control of hydroxyapatite crystals via hydrothermal synthesis. Crystal Growth and Design, 9, 466–474.CrossRefGoogle Scholar
  19. 19.
    Ma, M. G., & Zhu, J. F. (2009). Solvothermal synthesis and characterization of hierarchically nanostructured hydroxyapatite hollow spheres. European Journal of Inorganic Chemistry, 2009, 5522–5526.CrossRefGoogle Scholar
  20. 20.
    Bigi, A., Boanini, E., & Rubini, K. (2004). Hydroxyapatite gels and nanocrystals prepared through a sol–gel process. Journal of Solid State Chemistry, 177(9), 3092–3098.CrossRefGoogle Scholar
  21. 21.
    Kithva, P., Grøndahl, L., Martin, D., & Trau, M. (2010). Biomimetic synthesis and tensile properties of nanostructured high volume fraction hydroxyapatite and chitosan biocomposite films. Journal of Materials Chemistry, 20(2), 381–389.CrossRefGoogle Scholar
  22. 22.
    López-Macipe, A., Gómez-Morales, J., & Rodríguez-Clemente, R. (1998). Nanosized hydroxyapatite precipitation from homogeneous calcium/citrate/phosphate solutions using microwave and conventional heating. Advanced Materials, 10(1), 49–53.CrossRefGoogle Scholar
  23. 23.
    Wang, X., Qian, C., & Yu, X. (2014). Synthesis of nano-hydroxyapatite via microbial method and its characterization. Applied Biochemistry and Biotechnology, 173(4), 1003–1010.CrossRefGoogle Scholar
  24. 24.
    Barakat, N. A., Khalil, K., Sheikh, F. A., Omran, A., Gaihre, B., Khil, S. M., & Kim, H. Y. (2008). Physiochemical characterizations of hydroxyapatite extracted from bovine bones by three different methods: extraction of biologically desirable HAp. Materials Science and Engineering: C, 28(8), 1381–1387.CrossRefGoogle Scholar
  25. 25.
    Barakat, N. A., Khil, M. S., Omran, A., Sheikh, F. A., & Kim, H. Y. (2009). Extraction of pure natural hydroxyapatite from the bovine bones bio waste by three different methods. Journal of Materials Processing Technology, 209(7), 3408–3415.CrossRefGoogle Scholar
  26. 26.
    He, G., Dahl, T., Veis, A., & George, A. (2003). Nucleation of apatite crystals in vitro by self-assembled dentin matrix protein 1. Nature Materials, 2(8), 552–558.CrossRefGoogle Scholar
  27. 27.
    Antonakos, A., Liarokapis, E., & Leventouri, T. (2007). Micro-Raman and FTIR studies of synthetic and natural apatites. Biomaterials, 28(19), 3043–3054.CrossRefGoogle Scholar
  28. 28.
    Wei, G., Reichert, J. r., Bossert, J. r., & Jandt, K. D. (2008). Novel biopolymeric template for the nucleation and growth of hydroxyapatite crystals based on self-assembled fibrinogen fibrils. Biomacromolecules, 9(11), 3258–3267.CrossRefGoogle Scholar
  29. 29.
    Blakeslee, K., & Condrate, R. A. (1971). Vibrational spectra of hydrothermally prepared hydroxyapatites. Journal of the American Ceramic Society, 54(11), 559–563.CrossRefGoogle Scholar
  30. 30.
    Lee, Y., Hahm, Y. M., Matsuya, S., Nakagawa, M., & Ishikawa, K. (2007). Characterization of macroporous carbonate-substituted hydroxyapatite bodies prepared in different phosphate solutions. Journal of Materials Science, 42(18), 7843–7849.CrossRefGoogle Scholar
  31. 31.
    Lim, S.-R., Gooi, B.-H., Singh, M., & Gam, L.-H. (2011). Analysis of differentially expressed proteins in colorectal cancer using hydroxyapatite column and SDS-PAGE. Applied Biochemistry and Biotechnology, 165(5-6), 1211–1224.CrossRefGoogle Scholar
  32. 32.
    Maachou, H., Bal, K., Bal, Y., Chagnes, A., Cote, G., & Aliouche, D. (2012). In vitro biomineralization and bulk characterization of chitosan/hydroxyapatite composite microparticles prepared by emulsification cross-linking method: orthopedic use. Applied Biochemistry and Biotechnology, 168(6), 1459–1475.CrossRefGoogle Scholar
  33. 33.
    Lee, J. P., Lee, J. S., & Park, S. C. (1999). Two-phase methanization of food wastes in pilot scale. Twentieth Symposium on Biotechnology for Fuels and Chemicals (pp. 585–593). Berlin: Springer.CrossRefGoogle Scholar
  34. 34.
    Han, G., Shin, S. G., Lee, J., Lee, C., Jo, M., & Hwang, S. (2016). Mesophilic acidogenesis of food waste-recycling wastewater: effects of hydraulic retention time, pH, and temperature. Applied Biochemistry and Biotechnology, 180(5), 980–999.CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biology, Faculty of ScienceAlbaha UniversityAlbahaKingdom of Saudi Arabia

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