Polymer Bulletin

, Volume 76, Issue 2, pp 725–745 | Cite as

Synthesis, characterization and biocompatible properties of novel silk fibroin/graphene oxide nanocomposite scaffolds for bone tissue engineering application

  • Mehdi Narimani
  • Abbas TeimouriEmail author
  • Zeinab Shahbazarab
Original Paper


Novel three-dimensional porous silk fibroin/graphene oxide (SF/GO) nanocomposite scaffolds with different graphene oxide (GO) concentrations were prepared by using the freeze-drying technique. The obtained SF/GO scaffolds were characterized by thermogravimetric analysis, X-ray diffraction, scanning electron microscopy, Brunauer–Emmett–Teller isotherm and Fourier transform infrared spectroscopy techniques. The water absorption, compressive properties, porosity, degradation, biomineralization capability, cell attachment and cell viability of the composite scaffolds were studied as well. Cytocompatibility of the scaffolds was studied in vitro by employing the methylthiazoletetrazolium assay. The results showed that the presence of graphene oxide nanoparticles throughout the fibroin matrix led to an increase in water uptake and mechanical properties; at the same time, the porosity of the scaffolds was decreased. The cell adhesion results also indicated that human osteoblast cells (MG-63) could adhere to the surface of SF/GO nanocomposites and develop on them. These suggest that SF/GO nanocomposite scaffolds may be a good candidate for bone tissue engineering applications.


Composite scaffold Graphene oxide Silk fibroin Tissue engineering 



The authors are thankful from Payame Noor University in Isfahan Research Council (Grant # 68424), and contributions from Isfahan University of Technology are gratefully acknowledged.


  1. 1.
    Jayakumar R, Chennazhi KP, Srinivasan S, Nair SV, Furuike T, Tamura H (2011) Chitin scaffolds in tissue engineering. Int J Mol Sci 15:1876–1887CrossRefGoogle Scholar
  2. 2.
    Rocchietta I, Fontana F, Simion M (2008) Clinical outcomes of vertical bone augmentation to enable dental implant placement: a systematic review. J Clin Periodontol 35:203–215CrossRefGoogle Scholar
  3. 3.
    Wang M (2003) Developing bioactive composite materials for tissue replacement. Biomaterials 24:2133–2151CrossRefGoogle Scholar
  4. 4.
    Hutmacher DW (2000) Scaffolds in tissue engineering bone and cartilage. Biomaterials 21:2529–2543CrossRefGoogle Scholar
  5. 5.
    Yang S, Leong KF, Du Z, Chua CK (2001) The design of scaffolds for use in tissue engineering. Part I. Traditional factors. Tissue Eng 7:679–689CrossRefGoogle Scholar
  6. 6.
    Ryan G, Pandit A, Apatsidis DP (2006) Fabrication methods of porous metals for use in orthopaedic applications. Biomaterials 27:2651–2670CrossRefGoogle Scholar
  7. 7.
    Lee CH, Singla A, Lee Y (2001) Biomedical applications of collagen. Int J Pharm 221:1–22CrossRefGoogle Scholar
  8. 8.
    Bensaıd W, Triffitt JT, Blanchat C, Oudina K, Sedel L, Petite H (2003) A biodegradable fibrin scaffold for mesenchymal stem cell transplantation. Biomaterials 24:2497–2502CrossRefGoogle Scholar
  9. 9.
    Kim UJ, Park J, Kim HJ, Wada M, Kaplan DL (2005) Three-dimensional aqueous-derived biomaterial scaffolds from silk fibroin. Biomaterials 26:2775–2785CrossRefGoogle Scholar
  10. 10.
    Singh BN, Panda NN, Mund R, Pramanik K (2016) Carboxymethyl cellulose enables silk fibroin nanofibrous scaffold with enhanced biomimetic potential for bone tissue engineering application. Carbohydr Polym 151:335–347CrossRefGoogle Scholar
  11. 11.
    Meinel L, Karageorgiou V, Hofmann S, Fajardo R, Snyder B, Li C, Zichner L, Langer R, Vunjak-Novakovic G, Kaplan DL (2004) Engineering bone-like tissue in vitro using human bone marrow stem cells and silk scaffolds. J Biomed Mater Res 71:25–34CrossRefGoogle Scholar
  12. 12.
    Luangbudnark W, Viyoch J, Laupattarakasem W, Surakunprapha P, Laupattarakasem P (2012) Properties and biocompatibility of chitosan and silk fibroin blend films for application in skin tissue engineering. World J Sci 2012:697201–697211CrossRefGoogle Scholar
  13. 13.
    Bhardwaj N, Kundu SC (2011) Silk fibroin protein and chitosan polyelectrolyte complex porous scaffolds for tissue engineering applications. Carbohydr Polym 85:325–333CrossRefGoogle Scholar
  14. 14.
    She Z, Liu W, Feng Q (2010) Silk fibroin/chitosan/heparin scaffold: preparation, antithrombogenicity and culture with hepatocytes. Polym Int 59:55–61CrossRefGoogle Scholar
  15. 15.
    Ruiz ON, Fernando KS, Wang B, Brown NA, Luo PG, McNamara ND, Vangsness M, Sun YP, Bunker CE (2011) Graphene oxide: a nonspecific enhancer of cellular growth. ACS Nano 5:8100–8107CrossRefGoogle Scholar
  16. 16.
    Mehrali M, Moghaddam E, Shirazi SF, Baradaran S, Mehrali M, Latibari ST, Metselaar HS, Kadri NA, Zandi K, Osman NA (2014) Synthesis, mechanical properties, and in vitro biocompatibility with osteoblasts of calcium silicate–reduced graphene oxide composites. ACS Appl Mater Interface 6:3947–3962CrossRefGoogle Scholar
  17. 17.
    Huang G, Chen S, Tang S, Gao J (2012) A novel intumescent flame retardant-functionalized graphene: Nanocomposite synthesis, characterization, and flammability properties. Mater Chem Phys 135:938–947CrossRefGoogle Scholar
  18. 18.
    Kalbacova M, Broz A, Kong J, Kalbac M (2010) Graphene substrates promote adherence of human osteoblasts and mesenchymal stromal cells. Carbon 48:4323–4329CrossRefGoogle Scholar
  19. 19.
    Nayak TR, Andersen H, Makam VS, Khaw C, Bae S, Xu X, Ee PL, Ahn JH, Hong BH, Pastorin G, Ozyilmaz B (2011) Graphene for controlled and accelerated osteogenic differentiation of human mesenchymal stem cells. ACS Nano 5:4670–4678CrossRefGoogle Scholar
  20. 20.
    Zhang J, Zhang F, Yang H, Huang X, Liu H, Zhang J, Guo S (2010) Graphene oxide as a matrix for enzyme immobilization. Langmuir 26:6083–6085CrossRefGoogle Scholar
  21. 21.
    Sui ZY, Cui Y, Zhu JH, Han BH (2013) Preparation of three-dimensional graphene oxide–polyethylenimine porous materials as dye and gas adsorbents. ACS Appl Mater Interface 5:9172–9179CrossRefGoogle Scholar
  22. 22.
    Ruiz ON, Fernando KS, Wang B, Brown NA, Luo PG, McNamara ND, Vangsness M, Sun YP, Bunker CE (2011) Graphene oxide: a nonspecific enhancer of cellular growth. ACS Nano 5:8100–8107CrossRefGoogle Scholar
  23. 23.
    Yoon OJ, Jung CY, Sohn IY, Kim HJ, Hong B, Jhon MS, Lee NE (2011) Nanocomposite nanofibers of poly(d, l-lactic-co-glycolic acid) and graphene oxide nanosheets. Compos Part A Appl Sci Manuf 42:1978–1984CrossRefGoogle Scholar
  24. 24.
    Yoon OJ, Sohn IY, Kim DJ, Lee NE (2012) Enhancement of thermomechanical properties of poly (d, l-lactic-co-glycolic acid) and graphene oxide composite films for scaffolds. Macromol Res 1:1–6Google Scholar
  25. 25.
    Wu S, Zhao X, Cui Z, Zhao C, Wang Y, Du L, Li Y (2014) Cytotoxicity of graphene oxide and graphene oxide loaded with doxorubicin on human multiple myeloma cells. Int J Nano Med 9:1413–1421Google Scholar
  26. 26.
    Teimouri A, Ebrahimi R, Chermahini AN, Emadi R (2015) Fabrication and characterization of silk fibroin/chitosan/nano γ-alumina composite scaffolds for tissue engineering applications. RSC Adv 5:27558–27570CrossRefGoogle Scholar
  27. 27.
    Teimouri A, Ebrahimi R, Emadi R, Beni BH, Chermahini AN (2015) Nano-composite of silk fibroin–chitosan/Nano ZrO2 for tissue engineering applications: Fabrication and morphology. Int J Biol Macromol 76:292–302CrossRefGoogle Scholar
  28. 28.
    Ghorbanian L, Emadi R, Razavi SM, Shin H, Teimouri A (2013) Fabrication and characterization of novel diopside/silk fibroin nanocomposite scaffolds for potential application in maxillofacial bone regeneration. Int J Biol Macromol 58:275–280CrossRefGoogle Scholar
  29. 29.
    Rockwood DN, Preda RC, Yücel T, Wang X, Lovett ML, Kaplan DL (2011) Materials fabrication from Bombyx mori silk fibroin. Nat Protoc 6:1612–1631CrossRefGoogle Scholar
  30. 30.
    Lim HN, Huang NM, Lim SS, Harrison I, Chia CH (2011) Fabrication and characterization of graphene hydrogel via hydrothermal approach as a scaffold for preliminary study of cell growth. Int J Nano Med 6:1817–1823CrossRefGoogle Scholar
  31. 31.
    Kokubo T, Takadama H (2006) How useful is SBF in predicting in vivo bone bioactivity? Biomaterials 27:2907–2915CrossRefGoogle Scholar
  32. 32.
    Escamilla-García M, Calderon-Dominguez G, Chanona-Perez JJ, Farrera-Rebollo RR, Andraca-Adame JA, Arzate-Vazquez I, Mendez-Mendez JV, Moreno-Ruiz LA (2013) Physical and structural characterisation of zein and chitosan edible films using nanotechnology tools. Int J Biol Macromol 61:196–203CrossRefGoogle Scholar
  33. 33.
    Zdravkov B, Čermák J, Šefara M, Janků J (2007) Pore classification in the characterization of porous materials: a perspective. Open Chem 5:385–395CrossRefGoogle Scholar
  34. 34.
    Alonso-Lemus I, Verde-Gómez Y, Álvarez-Contreras L (2011) Platinum nanoparticles synthesis supported in mesoporous silica and its effect in MCM-41 lattice. Int J Electrochem Sci 6:4176–4187Google Scholar
  35. 35.
    Ho MH, Kuo PY, Hsieh HJ, Hsien TY, Hou LT, Lai JY, Wang DM (2004) Preparation of porous scaffolds by using freeze-extraction and freeze-gelation methods. Biomaterials 25:129–138CrossRefGoogle Scholar
  36. 36.
    Zeltinger J, Sherwood JK, Graham DA, Müeller R, Griffith LG (2001) Effect of pore size and void fraction on cellular adhesion, proliferation, and matrix deposition. Tissue Eng 7:557–572CrossRefGoogle Scholar
  37. 37.
    Wan Y, Chen X, Xiong G, Guo R, Luo H (2014) Synthesis and characterization of three-dimensional porous graphene oxide/sodium alginate scaffolds with enhanced mechanical properties. Mater Express 4:429–434CrossRefGoogle Scholar
  38. 38.
    Li J, Dou Y, Yang J, Yin Y, Zhang H, Yao F, Wang H, Yao K (2009) Surface characterization and biocompatibility of micro-and nano-hydroxyapatite/chitosan-gelatin network films. Mater Sci Eng 29:1207–1215CrossRefGoogle Scholar
  39. 39.
    Vacanti JP, Langer R (1999) Tissue engineering: the design and fabrication of living replacement devices for surgical reconstruction and transplantation. Lancet 354:32–34CrossRefGoogle Scholar
  40. 40.
    Thein-Han WW, Misra RD (2009) Biomimetic chitosan–nanohydroxyapatite composite scaffolds for bone tissue engineering. Acta Biomater 5:1182–1197CrossRefGoogle Scholar
  41. 41.
    Kumar PS, Srinivasan S, Lakshmanan VK, Tamura H, Nair SV, Jayakumar R (2011) Synthesis, characterization and cytocompatibility studies of α-chitin hydrogel/nano hydroxyapatite composite scaffolds. Int J Biol Macromol 49:20–31CrossRefGoogle Scholar
  42. 42.
    Liao KH, Lin YS, Macosko CW, Haynes CL (2011) Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts. ACS Appl Mater interface 3:2607–2615CrossRefGoogle Scholar
  43. 43.
    Song J, Gao H, Zhu G, Cao X, Shi X, Wang Y (2015) The preparation and characterization of polycaprolactone/graphene oxide biocomposite nanofiber scaffolds and their application for directing cell behaviors. Carbon 95:1039–1050CrossRefGoogle Scholar
  44. 44.
    Anselme K (2000) Osteoblast adhesion on biomaterials. Biomaterials 21:667–681CrossRefGoogle Scholar
  45. 45.
    Aliramaji S, Zamanian A, Mozafari M (2017) Super-paramagnetic responsive silk fibroin/chitosan/magnetite scaffolds with tunable pore structures for bone tissue engineering applications. Mater Sci Eng 70:736–744CrossRefGoogle Scholar
  46. 46.
    Xie C, Lu X, Han L, Xu J, Wang Z, Jiang L, Wang K, Zhang H, Ren F, Tang Y (2016) Biomimetic mineralized hierarchical graphene oxide/chitosan scaffolds with adsorbability for immobilization of nanoparticles for biomedical applications. ACS Appl Mater Interface 8:1707–1717CrossRefGoogle Scholar
  47. 47.
    Azadi M, Teimouri A, Mehranzadeh G (2016) Preparation, characterization and biocompatible properties of β-chitin/silk fibroin/nanohydroxyapatite composite scaffolds prepared using a freeze-drying method. RSC Adv 6:7048–7060CrossRefGoogle Scholar
  48. 48.
    She Z, Zhang B, Jin C, Feng Q, Xu Y (2008) Preparation and in vitro degradation of porous three-dimensional silk fibroin/chitosan scaffold. Polym Degrad Stab 93:1316–1322CrossRefGoogle Scholar
  49. 49.
    Bhardwaj N, Kundu SC (2011) Silk fibroin protein and chitosan polyelectrolyte complex porous scaffolds for tissue engineering applications. Carbohydr Polym 85:325–333CrossRefGoogle Scholar
  50. 50.
    Dinescu S, Ionita M, Pandele AM, Galateanu B, Iovu H, Ardelean A, Costache M, Hermenean A (2014) In vitro cytocompatibility evaluation of chitosan/graphene oxide 3D scaffold composites designed for bone tissue engineering. Bio-Med Mater Eng 24:2249–2256Google Scholar
  51. 51.
    Wang L, Li C (2007) Preparation and physicochemical properties of a novel hydroxyapatite/chitosan–silk fibroin composite. Carbohydr Polym 68:740–745CrossRefGoogle Scholar
  52. 52.
    Kim HS, Kim JT, Jung YJ, Ryu SC, Son HJ, Kim YG (2007) Preparation of a porous chitosan/fibroin-hydroxyapatite composite matrix for tissue engineering. Macromol Res 15:65–73CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mehdi Narimani
    • 1
  • Abbas Teimouri
    • 1
    Email author
  • Zeinab Shahbazarab
    • 1
  1. 1.Department of ChemistryPayame Noor University (PNU)TehranIran

Personalised recommendations