Journal of the Australian Ceramic Society

, Volume 55, Issue 1, pp 269–279 | Cite as

Biocompatibility of a PLA-based composite containing hydroxyapatite derived from waste bones of dolphin Neophocaena asiaeorientalis

  • Mi Rim Lee
  • Gyung Won Lee
  • Ji Eun Kim
  • Woo Bin Yun
  • Jun Young Choi
  • Jin Ju Park
  • Hye Ryeong Kim
  • Bo Ram Song
  • Ji Won Park
  • Mi Ju Kang
  • Yong Rock Ann
  • Jung Youn Park
  • Seung Yun Yang
  • Dae Youn HwangEmail author


Natural hydroxyapatite (HA), derived from waste bones of several animal species, has received much attention as a material for bone grafts and fillers and has a role as a coating for metal implants because of its biocompatibility and non-toxicity. To investigate the applicability of HA derived from waste bones of novel animal sources, the biocompatibility and toxicity of a poly-l-lactic acid (PLA)-based composite containing HA derived from the backbone of the dolphin Neophocaena asiaeorientalis (HANA) were examined in Sprague-Dawley (SD) rats. HANA powder showed X-ray diffraction peak patterns that corresponded to those of standard HA. Among five composites prepared from different combinations of PLA and HANA (7:3, 6:4, 5:5, 4:6, and 3:7), a PLA/HANA composite manufactured with a 6:4 PLA:HANA ratio had high surface roughness (453 nm), 10.3 N of maximum load, and 451.9 MPa of module elasticity. After implantation in the subcutaneous region of SD rats for 8 weeks, the amount of confluent, aggregated structures of multilayered cells on the PLA/HANA implant surface was greater than that on the PLA surface, although both implants were completely covered with adhesive cells. During the implant period, the initial intact form of the PLA/HANA composite broke into small fragments with few inflammatory cells in the contact region and no indication of significant toxicity. Taken together, the results suggest that HANA may have good biocompatibility and be non-toxic as it did not induce an immune response in SD rats.


Hydroxyapatite Dolphin Neophocaena asiaeorientalis Waste bone PLA composite Biocompatibility 



We thank Jin Hyang Hwang, the animal technician, for directing the care and management of animals at the Laboratory Animal Resources Center in Pusan National University.


This study was supported by grants to Professor Dae Youn Hwang from the Korea Institute of Planning & Evaluation for Technology in Food, Agriculture and Forestry (116027-032-HD030) and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1D1A3B03032631).


  1. 1.
    Elvevoll, E.O., Sørensen, N.K., Osterud, B., Ofstad, R., Martinez, I.: Processing of marine foods. Meat Sci. (1996).
  2. 2.
    Gosho, M.E., Rice, D.W., Breiwick, J.M.: The sperm whale, Physeter microcephalus. Mar Fish Rev. 46, 54–64 (1984)Google Scholar
  3. 3.
    Zelko, F.: From blubber and baleen to buddha of the deep: The rise of the metaphysical whale. Soc Anim. (2012).
  4. 4.
    Ozawa, M., Suzuki, S.: Microstructural development of natural hydroxyapatite originated from fish-bone waste through heat treatment. J Am Ceram Soc. (2002).
  5. 5.
    Higgs, N.D., Little, C.T., Glover, A.G.: Bones as biofuel: a review of whale bone composition with implications for deep-sea biology and palaeoanthropology. Proc Biol Sci. (2011).
  6. 6.
    Nemliher, J.G., Baturin, G.N., Kallaste, T.E., Murdmaa, I.O.: Transformation of hydroxyapatite of bone phosphate from the ocean bottom during fossilization. Lithol Miner Resour. (2004).
  7. 7.
    Pandharipande, S., Sondawale, S.: Review on synthesis methods of hydroxyapatite and its biocomposites. Int J Sci Res Eng Technol. 17, 3410–3416 (2016)Google Scholar
  8. 8.
    Elliott, J.C.: Structure and chemistry of the apatites and other calcium orthophosphates, Studies in Inorganic Chemistry, pp. 191–301. Elsevier, Amsterdam (1994)Google Scholar
  9. 9.
    Hulber, S.F., Bokros, J.C., Hench, L.L., Wilson, J., Heimke, G.: Ceramics in clinical applications: past, present and future. In: Vincenzini, P. (ed.) High Tech Ceramics, pp. 189–213. Elsevier, Amsterdam (1987)Google Scholar
  10. 10.
    Joschek, S., Nies, B., Krotz, R., Göpferich, A.: Chemical and physicochemical characterization of porous hydroxyapatite ceramics made of natural bone. Biomaterials. (2000).
  11. 11.
    Lee, C.K., Choi, J.S., Jeon, Y.J., Byun, H.G., Kim, S.K.: The properties of natural hydroxyapatite isolated from tuna bone. J Kor Fish Soc. 30, 652–659 (1997)Google Scholar
  12. 12.
    Xiaoying, L., Yongbin, F., Dachun, G., Wei, C.: Preparation and characterization of natural hydroxyapatite from animal hard tissues. Key Eng Mater. (2007).
  13. 13.
    Barakata, N.A.M., Khila, M.S., Omrand, A.M., Sheikhd, F.A., Kim, H.Y.: Extraction of pure natural hydroxyapatite from the bovine bones bio waste by three different methods. J Mat Proc Technol. (2009).
  14. 14.
    Haberko, K., Bucko, M.M., Brzezinska-Miecznik, J., Haberko, M., Mozgawa, W., Panz, T., Pyda, A., Zarebski, J.: Natural hydroxyapatite—its behaviour during heat treatment. J Eur Cer Soci. (2006).
  15. 15.
    Hellmich, C., Ulm, F.J.: Average hydroxyapatite concentration is uniform in the extracollagenous ultrastructure of mineralized tissues: evidence at the 1-10­μm scale. Biomechan Model Mechanobiol. (2003).
  16. 16.
    Kim, J.W., Kim, H.S.: Synthesis and characteristics of poly(l-lactic acid-block-γ-aminobutyric acid). Text Sci Eng. (2015).
  17. 17.
    JCPDS Card No. 9–432, 1996Google Scholar
  18. 18.
    Song, S.H., Kim, J.E., Lee, Y.J., Kwak, M.H., Sung, G.Y., Kwon, S.H., Son, H.J., Lee, H.S., Jung, Y.J., Hwang, D.Y.: Cellulose film regenerated from Styela clava tunics have biodegradability, toxicity and biocompatibility in the skin of SD rats. J Mater Sci Mater Med. (2014).
  19. 19.
    Seong, K.Y., Koh, E.K., Lee, S.H., Kwak, M.H., Son, H.J., Lee, H.S., Hwang, D.Y., Jung, Y.J.: Preparation and characterization of high absorptive cellulose film derived from Styela Clava tunic for wound dressing. Text Coloration Finish. (2015).
  20. 20.
    Palacio, M.L.B., Bhushan, B.: Bioadhesion: a review of concepts and applications. Phil Trans R Soc A. (2018).
  21. 21.
    Alvarez-Barreto, J.F., Landy, B., VanGordon, S., Place, L., DeAngelis, P.L., Sikavitsas, V.I.: Enhanced osteoblastic differentiation of mesenchymal stem cells seeded in RGD-functionalized PLLA scaffolds and cultured in a flow perfusion bioreactor. J Tissue Eng Regen Med. (2011).
  22. 22.
    Macha, I.J., Ben-Nissan, B., Santos, J., Cazalbou, S., Stamboulis, A., Grossin, D., Giordano, G.: Biocompatibility of a new biodegradable polymer-hydroxyapatite composite for biomedical applications. J Drug Deliv Sci Technol. (2017).
  23. 23.
    Thevenot, P., Hu, W., Tang, L.: Surface chemistry influences implant biocompatibility. Curr Top Med Chem. (2008).
  24. 24.
    García-Gareta, E., Coathup, M.J., Blunn, G.W.: Osteoinduction of bone grafting materials for bone repair and regeneration. Bone. (2015).
  25. 25.
    Tayton, E., Purcell, M., Aarvold, A., Smith, J.O., Briscoe, A., Kanczler, J.M., Shakesheff, K.M., Howdle, S.M., Dunlop, D.G., Oreffo, R.O.: A comparison of polymer and polymer-hydroxyapatite composite tissue engineered scaffolds for use in bone regeneration. An in vitro and in vivo study. J Biomed Mater Res A. (2014).
  26. 26.
    Russias, J., Saiz, E., Nalla, R.K., Gryn, K., Ritchie, R.O., Tomsia, A.P.: Fabrication and mechanical properties of PLA/HA composites: a study of in vitro degradation. Mater Sci Eng C Biomim Supramol Syst. (2006).
  27. 27.
    Barbieri, D., Renard, A.J., de Bruijn, J.D., Yuan, H.: Heterotopic bone formation by nano-apatite containing poly(D,L-lactide) composites. Eur Cell Mater. 19, 252–261 (2010)CrossRefGoogle Scholar
  28. 28.
    Zong, C., Qian, X., Tang, Z., Hu, Q., Chen, J., Gao, C., Tang, R., Tong, X., Wang, J.: Biocompatibility and bone-repairing effects: comparison between porous poly-lactic-co-glycolic acid and nano-hydroxyapatite/poly(lactic acid) scaffolds. J Biomed Nanotechnol. (2014).
  29. 29.
    Canoux, C.B., Barbieri, D., Yuan, H., de Bruijn, J.D., van Blitterswijk, C.A., Habibovic, P.: In vitro and in vivo bioactivity assessment of a polylactic acid/hydroxyapatite composite for bone regeneration. Biomatter. (2014).
  30. 30.
    Rizzi, S.C., Heath, D.J., Coombes, A.G., Bock, N., Textor, M., Downes, S.: Biodegradable polymer/hydroxyapatite composites: surface analysis and initial attachment of human osteoblasts. J Biomed Mater Res. (2001).<475::AID-JBM1039>3.0.CO;2-Q
  31. 31.
    Wojcieszak, D., Mazur, M., Kalisz, M., Grobelny, M.: Influence of Cu, Au and Ag on structural and surface properties of bioactive coatings based on titanium. Master Sci Eng C. (2017).
  32. 32.
    Khan, S.P., Auner, G.G., Newaz, G.M.: Influence of nanoscale surface roughness on neural cell attachment on silicon. Nanomedicine. (2005).
  33. 33.
    Keshel, S.H., Azhdadi, S.N., Asefnejad, A., Sadraeian, M., Montazeri, M., Biazar, E.: The relationship between cellular adhesion and surface roughness for polyurethane modified by microwave plasma radiation. Int J Nanomedicine. (2011).

Copyright information

© Australian Ceramic Society 2018

Authors and Affiliations

  • Mi Rim Lee
    • 1
  • Gyung Won Lee
    • 1
  • Ji Eun Kim
    • 1
  • Woo Bin Yun
    • 1
  • Jun Young Choi
    • 1
  • Jin Ju Park
    • 1
  • Hye Ryeong Kim
    • 1
  • Bo Ram Song
    • 1
  • Ji Won Park
    • 1
  • Mi Ju Kang
    • 1
  • Yong Rock Ann
    • 2
  • Jung Youn Park
    • 3
  • Seung Yun Yang
    • 1
  • Dae Youn Hwang
    • 1
    Email author
  1. 1.Department of Biomaterials Science, College of Natural Resources & Life Science/Life and Industry Convergence Research InstitutePusan National UniversityMiryang-siSouth Korea
  2. 2.National Marine Biodiversity Institute of KoreaChungcheongnam-doSouth Korea
  3. 3.National Institute of Fisheries ScienceGijang-gunSouth Korea

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