Journal of Nanoparticle Research

, 14:1265

In vitro and in vivo investigations on bone regeneration potential of laminated hydroxyapatite/gelatin nanocomposite scaffold along with DBM

  • Shima Tavakol
  • Iraj Ragerdi Kashani
  • Mahmood Azami
  • Ahad Khoshzaban
  • Behnaz Tavakol
  • Sharmin Kharrazi
  • Somayeh Ebrahimi
  • Seyed Mahdi Rezayat Sorkhabadi
Research Paper

Abstract

Bone regeneration ability of a scaffold strongly depends on its structure and the size of its components. In this study, a nanostructured scaffold was designed for bone repair using nano hydroxyapatite (nHA) (8–16 nm × 50–80 nm) and gelatin (GEL) as main components. In vitro investigations of calcium matrix deposition and gene expression of the seeded cells for this scaffold, demineralized bone matrix (DBM), scaffold plus DBM, and the control group were carried out. Bone regeneration in rat calvarium with critical defect size after 1, 4, and 8 weeks post implantation was investigated. The calcium matrix depositions by the osteoblast and RUNX2, ALP, osteonectin, and osteocalcin gene expression in scaffold were more significant than in other groups. Histomorphometry analysis confirmed in vitro results. In vitro and in vivo bone regeneration were least in scaffold plus DBM group. Enhanced effects in scaffold could be attributed to the shape and size of nHA particles and good architecture of the scaffold. Reduction of bone regeneration might be due to tight bonding of BMPs and nHA particles in the third group. Results obtained from this study confirmed that nano-scale size of the main components and the scaffold architecture (pore diameter, interconnectivity pores, etc.) have significant effects on bone regeneration ability of the scaffold and are important parameters in designing a temporary bone substitute.

Keywords

Scaffold Nano hydroxyapatite Demineralized bone matrix Bone regeneration Gene expression Histomorphometry 

References

  1. Abdel-Gawad EI, Awwad S (2010) Biocompatibility of intravenous nano hydroxyapatite in male rats. Nat Sci 8:60–68Google Scholar
  2. Akay G, Birch MA et al (2004) Microcellular polyHIPE polymer supports osteoblast growth and bone formation in vitro. Biomaterials 25(18):3991–4000CrossRefGoogle Scholar
  3. Alper G, Bernick SOL et al (1989) Osteogenesis in bone defects in rats: the effects of hydroxyapatite and demineralized bone matrix. Am J Med Sci 298(6):371CrossRefGoogle Scholar
  4. Aronow MA, Gerstenfeld LC et al (1990) Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells. J Cell Physiol 143(2):213–221CrossRefGoogle Scholar
  5. Askarzadeh K, Orang F et al (2005) Fabrication and characterization of a porous composite scaffold based on gelatin and hydroxyapatite for bone tissue engineering. Iran Polym J 14(6):511–520Google Scholar
  6. Azami M, Samadikuchaksaraei A et al (2010) Synthesis and characterization of a laminated hydroxyapatite/gelatin nanocomposite scaffold with controlled pore structure for bone tissue engineering. Int J Artif Organs 33(2):86–95Google Scholar
  7. Catledge SA, Fries MD et al (2002) Nanostructured ceramics for biomedical implants. J Nanosci Nanotechnol 23(4):293–312CrossRefGoogle Scholar
  8. Chen L, Joseph MM, James C-ML, Hao L (2011) The role of surface charge on the uptake and biocompatibility of hydroxyapatite nanoparticles with osteoblast cells. Nanotechnology 22(10):105708CrossRefGoogle Scholar
  9. Dallari D, Savarino L et al (2012) A prospective, randomised, controlled trial using a Mg-hydroxyapatite-demineralized bone matrix nanocomposite in tibial osteotomy. Biomaterials 33(1):72–79CrossRefGoogle Scholar
  10. Dvorak MM, Riccardi D (2004) Ca2+ as an extracellular signal in bone. Cell Calcium 35(3):249–255CrossRefGoogle Scholar
  11. George J, Kuboki Y et al (2006) Differentiation of mesenchymal stem cells into osteoblasts on honeycomb collagen scaffolds. Biotechnol Bioeng 95(3):404–411CrossRefGoogle Scholar
  12. Götz W, Lenz S, Reichert C, Kai-Olaf et al (2010) A preliminary study in osteoinduction by a nano-crystalline hydroxyapatite in the mini pig. Folia Histochem Cytobiol 48:589–596CrossRefGoogle Scholar
  13. Hassenkam T, Fantner GE et al (2004) High-resolution AFM imaging of intact and fractured trabecular bone. Bone 35(1):4–10CrossRefGoogle Scholar
  14. Hu J, Liu ZS et al (2007) Effect of hydroxyapatite nanoparticles on the growth and p53/c-Myc protein expression of implanted hepatic VX ~ 2 tumor in rabbits by intravenous injection. World J Gastroenterol 13(20):2798Google Scholar
  15. Karageorgiou V, Kaplan D (2005) Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 26(27):5474–5491CrossRefGoogle Scholar
  16. Kartsogiannis V, Ng KW (2004) Cell lines and primary cell cultures in the study of bone cell biology. Mol Cell Endocrinol 228(1–2):79–102CrossRefGoogle Scholar
  17. Kim HW, Kim HE et al (2005) Stimulation of osteoblast responses to biomimetic nanocomposites of gelatin-hydroxyapatite for tissue engineering scaffolds. Biomaterials 26(25):5221–5230CrossRefGoogle Scholar
  18. Laurencin CT, Ambrosio AMA et al (1999) Tissue engineering: orthopedic applications. Annu Rev Biomed Eng 1(1):19–46CrossRefGoogle Scholar
  19. Liao S, Tamura K et al (2006) Human neutrophils reaction to the biodegraded nano hydroxyapatite/collagen and nano hydroxyapatite/collagen/poly (l-lactic acid) composites. J Biomed Mater Res A 76(4):820–825Google Scholar
  20. Liu Y, Lu Y et al (2009) Segmental bone regeneration using an rhBMP-2-loaded gelatin/nanohydroxyapatite/fibrin scaffold in a rabbit model. Biomaterials 30(31):6276–6285CrossRefGoogle Scholar
  21. Maeno S, Niki Y et al (2005) The effect of calcium ion concentration on osteoblast viability, proliferation and differentiation in monolayer and 3D culture. Biomaterials 26(23):4847–4855CrossRefGoogle Scholar
  22. Mobini S, Javadpour J et al (2008) Synthesis and characterisation of gelatinnano hydroxyapatite composite scaffolds for bone tissue engineering. Adv Appl Ceram 107(1):4–8CrossRefGoogle Scholar
  23. Okamoto Y, Horisaka Y et al (1991) Muscle tissue reactions to implantation of bone matrix gelatin. Clin Orthop Relat Res 263:242Google Scholar
  24. Olszta MJ, Cheng X et al (2007) Bone structure and formation: a new perspective. Mater Sci Eng, R 58(3–5):77–116CrossRefGoogle Scholar
  25. Otsuki B, Takemoto M, Fujibayashi S, Neo M, Kokubo T, Nakamura T (2006) Pore throat size and connectivity determine bone and tissue ingrowth into porous implants: three dimensional micro-CT based structural analyses of porous bioactive titanium implants. Biomaterials 27(35):5892–5900CrossRefGoogle Scholar
  26. Papay FA, Morales L Jr et al (1996) Comparison of ossification of demineralized bone, hydroxyapatite, Gelfoam, and bone wax in cranial defect repair. J Craniofac Surg 7(5):347CrossRefGoogle Scholar
  27. Petit R (1999) The use of hydroxyapatite in orthopaedic surgery: a ten-year review. Euro J Ortho Surg Trauma 9(2):71–74CrossRefGoogle Scholar
  28. Pezzatini S, Solito R et al (2006) The effect of hydroxyapatite nanocrystals on microvascular endothelial cell viability and functions. J Biomed Mater Res A 76(3):656–663Google Scholar
  29. Phillips MJ, Darr JA et al (2003) Synthesis and characterization of nano-biomaterials with potential osteological applications. J Mater Sci Mater Med 14(10):875–882CrossRefGoogle Scholar
  30. Pi M, Faber P et al (2005) Identification of a novel extracellular cation-sensing G-protein-coupled receptor. J Biol Chem 280(48):40201CrossRefGoogle Scholar
  31. Rabie ABM, Lie Ken Jie RKP (1996) Integration of endochondral bone grafts in the presence of demineralized bone matrix. Int J Oral Maxillofac Surg 25(4):311–318CrossRefGoogle Scholar
  32. Saunders R, Szymczyk KH et al (2007) Matrix regulation of skeletal cell apoptosis III: mechanism of ion pair induced apoptosis. J Cell Biochem 100(3):703–715CrossRefGoogle Scholar
  33. Sicchieri LG, Crippa GE et al (2012) Pore size regulates cell and tissue interactions with PLGA-CaP scaffolds used for bone engineering. J Tissue Eng Regen Med 6(2):155–162CrossRefGoogle Scholar
  34. Sikavitsas VI, Temenoff JS et al (2001) Biomaterials and bone mechanotransduction. Biomaterials 22(19):2581–2593CrossRefGoogle Scholar
  35. Takaoka K, Nakahara H et al (1988) Ectopic bone induction on and in porous hydroxyapatite combined with collagen and bone morphogenetic protein. Clin Orthop Relat Res 234:250Google Scholar
  36. Tang R, Wang L et al (2004) Dissolution at the nanoscale: self preservation of biominerals. Angew Chem 116(20):2751–2755CrossRefGoogle Scholar
  37. Thein-Han WW, Misra RDK (2009) Three-dimensional chitosan-nanohydroxyapatite composite scaffolds for bone tissue engineering. JOM 61(9):41–44CrossRefGoogle Scholar
  38. Webster TJ (2003) Nanophase ceramics as improved bone tissue engineering materials. Am Ceram Soc Bull 82(6):23–28Google Scholar
  39. Woodard JR, Hilldore AJ et al (2007) The mechanical properties and osteoconductivity of hydroxyapatite bone scaffolds with multi-scale porosity. Biomaterials 28(1):45–54CrossRefGoogle Scholar
  40. Zhao Y, Zhang Y et al (2007) Synthesis and cellular biocompatibility of two kinds of HAP with different nanocrystal morphology. J Biomed Mater Res B Appl Biomater 83(1):121–126Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Shima Tavakol
    • 1
  • Iraj Ragerdi Kashani
    • 2
  • Mahmood Azami
    • 3
  • Ahad Khoshzaban
    • 4
  • Behnaz Tavakol
    • 5
  • Sharmin Kharrazi
    • 1
  • Somayeh Ebrahimi
    • 6
  • Seyed Mahdi Rezayat Sorkhabadi
    • 1
    • 7
    • 8
  1. 1.Department of Medical NanotechnologySchool of Advanced Technologies in Medicine, Tehran University of Medical SciencesTehranIran
  2. 2.Department of AnatomySchool of Medicine, Tehran University of Medical SciencesTehranIran
  3. 3.Department of Tissue EngineeringSchool of Advanced Technologies in Medicine, Tehran University of Medical SciencesTehranIran
  4. 4.Iranian Tissue Bank Research & Preparation CenterTehran University of Medical SciencesTehranIran
  5. 5.Department of MedicineKashan University of Medical SciencesKashanIran
  6. 6.Department of Biology, Faculty of SciencesUniversity of Tarbiat MoallemTehranIran
  7. 7.Department of Toxicology & Pharmacology, Faculty of PharmacyPharmaceutical Sciences Branch, Islamic Azad University (IAUPS)TehranIran
  8. 8.Department of Pharmacology, Faculty of MedicineTehran University of Medical SciencesTehranIran

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