Electrophoretic-deposited novel ternary silk fibroin/graphene oxide/hydroxyapatite nanocomposite coatings on titanium substrate for orthopedic applications

Abstract

The combination of graphene oxide (GO) with robust mechanical property, silk fibroin (SF) with fascinating biological effects and hydroxyapatite (HA) with superior osteogenic activity is a competitive approach to make novel coatings for orthopedic applications. Herein, the feasibility of depositing ternary SF/GO/HA nanocomposite coatings on Ti substrate was firstly verified by exploiting electrophoretic nanotechnology, with SF being used as both a charging additive and a dispersion agent. The surface morphology, microstructure and composition, in vitro hemocompatibility and in vitro cytocompatibility of the resulting coatings were investigated by SEM, Raman, FTIR spectra and biocompatibility tests. Results demonstrated that GO, HA and SF could be co-deposited with a uniform, smooth thin-film morphology. The hemolysis rate analysis and the platelet adhesion test indicated good blood compatibility of the coatings. The human osteosarcoma MG63 cells displayed well adhesion and proliferation behaviors on the prepared coatings, with enhanced ALP activities. The present study suggested that SF/GO/HA nanocomposite coatings could be a promising candidate for the surface functionalization of biomaterials, especially as orthopedic implant coating.

Keywords

graphene oxide / silk fibroin / hydroxyapatite / nano-biocomposite coatings / electrodeposition

References

[1] Kyu L S, Sup S B. Graphene: an emerging material for biological tissue engineering. Carbon Letters, 2013, 14(2): 63–75

[2] Goenka S, Sant V, Sant S. Graphene-based nanomaterials for drug delivery and tissue engineering. Journal of Controlled Release, 2014, 173: 75–88

[3] Wang X M, Zhang W H. Application of graphene derivatives in cancer therapy: a review. New Carbon Materials, 2013, 28(5): 321–326

[4] Kuila T, Bose S, Khanra P, . Recent advances in graphene-based biosensors. Biosensors & Bioelectronics, 2011, 26(12): 4637–4648

[5] Shen J, Zhu Y, Yang X, . Graphene quantum dots: emergent nanolights for bioimaging, sensors, catalysis and photovoltaic devices. Chemical Communications, 2012, 48(31): 3686–3699

[6] Nayak T R, Andersen H, Makam V S, . Graphene for controlled and accelerated osteogenic differentiation of human mesenchymal stem cells. ACS Nano, 2011, 5(6): 4670–4678

[7] Crowder S W, Prasai D, Rath R, . Three-dimensional graphene foams promote osteogenic differentiation of human mesenchymal stem cells. Nanoscale, 2013, 5(10): 4171–4176

[8] Lee W C, Lim C H, Shi H, . Origin of enhanced stem cell growth and differentiation on graphene and graphene oxide. ACS Nano, 2011, 5(9): 7334–7341

[9] Yoon H H, Bhang S H, Kim T, . Dual roles of graphene oxide in chondrogenic differentiation of adult stem cells: cell-adhesion substrate and growth factor-delivery carrier. Advanced Functional Materials, 2014, 24(41): 6455–6464

[10] Kim J, Choi K S, Kim Y, . Bioactive effects of graphene oxide cell culture substratum on structure and function of human adipose-derived stem cells. Journal of Biomedical Materials Research Part A, 2013, 101(12): 3520–3530

[11] Ku S H, Park C B. Myoblast differentiation on graphene oxide. Biomaterials, 2013, 34(8): 2017–2023

[12] Lee T J, Park S, Bhang S H, . Graphene enhances the cardiomyogenic differentiation of human embryonic stem cells. Biochemical and Biophysical Research Communications, 2014, 452(1): 174–180

[13] Park J, Park S, Ryu S, . Graphene-regulated cardiomyogenic differentiation process of mesenchymal stem cells by enhancing the expression of extracellular matrix proteins and cell signaling molecules. Advanced Healthcare Materials, 2014, 3(2): 176–181

[14] Li N, Zhang Q, Gao S, . Three-dimensional graphene foam as a biocompatible and conductive scaffold for neural stem cells. Scientific Reports, 2013, 3: 1604

[15] Wang Y, Kim H J, Vunjak-Novakovic G, . Stem cell-based tissue engineering with silk biomaterials. Biomaterials, 2006, 27(36): 6064–6082

[16] Elia R, Michelson C D, Perera A L, . Silk electrogel coatings for titanium dental implants. Journal of Biomaterials Applications, 2015, 29(9): 1247–1255

[17] Mandal B B, Grinberg A, Gil E S, . High-strength silk protein scaffolds for bone repair. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(20): 7699–7704

[18] Huang L, Li C, Yuan W, . Strong composite films with layered structures prepared by casting silk fibroin‒graphene oxide hydrogels. Nanoscale, 2013, 5(9): 3780–3786

[19] Ling S J, Li C, Adamcik J, . Directed growth of silk nanofibrils on graphene and their hybrid nanocomposites. ACS Macro Letters, 2014, 3(2): 146–152

[20] Hu K, Gupta M K, Kulkarni D D, . Ultra-robust graphene oxide‒silk fibroin nanocomposite membranes. Advanced Materials, 2013, 25(16): 2301–2307

[21] Xu S, Yong L, Wu P. One-pot, green, rapid synthesis of flowerlike gold nanoparticles/reduced graphene oxide composite with regenerated silk fibroin as efficient oxygen reduction electrocatalysts. ACS Applied Materials & Interfaces, 2013, 5(3): 654–662

[22] Yun Y S, Cho S Y, Jin H J. Carbon aerogels based on regenerated silk proteins and graphene oxide for supercapacitors. Macromolecular Research, 2014, 22(5): 509–514

[23] Liang B, Fang L, Hu Y, . Fabrication and application of flexible graphene silk composite film electrodes decorated with spiky Pt nanospheres. Nanoscale, 2014, 6(8): 4264–4274

[24] Mannoor M S, Tao H, Clayton J D, . Graphene-based wireless bacteria detection on tooth enamel. Nature Communications, 2012, 3(3): 132–136

[25] Paital S R, Dahotre N B. Calcium phosphate coatings for bio-implant applications: Materials, performance factors, and methodologies. Materials Science and Engineering R: Reports, 2009, 66(1‒3): 1–70

[26] Boccaccini A R, Keim S, Ma R, . Electrophoretic deposition of biomaterials. Journal of the Royal Society Interface, 2010, 7(Suppl 5): S581–S613

[27] Eliaz N, Sridhar T M, Kamachi Mudali U, . Electrochemical and electrophoretic deposition of hydroxyapatite for orthopaedic applications. Surface Engineering, 2005, 21(3): 238–242

[28] Chavez-Valdez A, Shaffer M S P, Boccaccini A R. Applications of graphene electrophoretic deposition. A review. The Journal of Physical Chemistry B, 2013, 117(6): 1502–1515

[29] Elia R, Michelson C D, Perera A L, . Electrodeposited silk coatings for bone implants. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2015, 103(8): 1602–1609

[30] Maniglio D, Bonani W, Bortoluzzi G, . Electrodeposition of silk fibroin on metal substrates. Journal of Bioactive and Compatible Polymers, 2010, 25(5): 441–454

[31] Li M, Liu Q, Jia Z, . Graphene oxide/hydroxyapatite composite coatings fabricated by electrophoretic nanotechnology for biological applications. Carbon, 2014, 67: 185–197

[32] Li M, Liu Q, Jia Z, . Electrophoretic deposition and electrochemical behavior of novel graphene oxide‒hyaluronic acid‒hydroxyapatite nanocomposite coatings. Applied Surface Science, 2013, 284: 804–810

[33] Shi Y Y, Li M, Liu Q, . Electrophoretic deposition of graphene oxide reinforced chitosan‒hydroxyapatite nanocomposite coatings on Ti substrate. Journal of Materials Science: Materials in Medicine, 2016, 27(3): 48

[34] Kweon H, Lee S W, Hahn B D, . Hydroxyapatite and silk combination-coated dental implants result in superior bone formation in the peri-implant area compared with hydroxyapatite and collagen combination-coated implants. Journal of Oral and Maxillofacial Surgery, 2014, 72(10): 1928–1936

[35] Rockwood D N, Preda R C, Yücel T, . Materials fabrication from Bombyx mori silk fibroin. Nature Protocols, 2011, 6(10): 1612–1631

[36] Li M, Cheng Y, Zheng Y F, . Surface characteristics and corrosion behaviour of WE43 magnesium alloy coated by SiC film. Applied Surface Science, 2012, 258(7): 3074–3081

[37] Li M, Wang Y, Liu Q, . In situ synthesis and biocompatibility of nano hydroxyapatite on pristine and chitosan functionalized graphene oxide. Journal of Materials Chemistry B: Materials for Biology and Medicine, 2013, 1(4): 475–484

[38] Matsumoto A, Chen J, Collette A L, . Mechanisms of silk fibroin sol-gel transitions. The Journal of Physical Chemistry B, 2006, 110(43): 21630–21638

[39] Supová M. Problem of hydroxyapatite dispersion in polymer matrices: a review. Journal of Materials Science: Materials in Medicine, 2009, 20(6): 1201–1213

[40] Li A B, Kluge J A, Guziewicz N A, . Silk-based stabilization of biomacromolecules. Journal of Controlled Release, 2015, 219: 416–430

[41] Kudin K N, Ozbas B, Schniepp H C, . Raman spectra of graphite oxide and functionalized graphene sheets. Nano Letters, 2008, 8(1): 36–41

[42] Koutsopoulos S. Synthesis and characterization of hydroxyapatite crystals: a review study on the analytical methods. Journal of Biomedical Materials Research, 2002, 62(4): 600–612

[43] Monti P, Taddei P, Freddi G, . Raman spectroscopic characterization of Bombyx mori silk fibroin: Raman spectrum of Silk I. Journal of Raman Spectroscopy, 2001, 32(2): 103–107

[44] Monti P, Freddi G, Tsukada M, . Spectroscopic characterization of Bombyx mori silk fibroin: Raman spectrum of Silk I. In: Greve J, Puppels G J, Otto C, ., eds. Spectroscopy of Biological Molecules: New Directions. Kluwer Academic Publisher, 1999, 81–82

[45] Arai T, Freddi G, Innocenti R, . Biodegradation of Bombyx mori silk fibroin fibers and films. Journal of Applied Polymer Science, 2004, 91(4): 2383–2390

[46] Bhumiratana S, Grayson W L, Castaneda A, . Nucleation and growth of mineralized bone matrix on silk‒hydroxyapatite composite scaffolds. Biomaterials, 2011, 32(11): 2812–2820

[47] Borkner C B, Elsner M B, Scheibel T. Coatings and films made of silk proteins. ACS Applied Materials & Interfaces, 2014, 6(18): 15611–15625

[48] Kim H J, Kim U J, Vunjak-Novakovic G, . Influence of macroporous protein scaffolds on bone tissue engineering from bone marrow stem cells. Biomaterials, 2005, 26(21): 4442–4452

[49] Erakovic S, Jankovic A, Tsui G C, . Novel bioactive antimicrobial lignin containing coatings on titanium obtained by electrophoretic deposition. International Journal of Molecular Sciences, 2014, 15(7): 12294–12322

[50] F756-00 A. Standard Practice for Assessment of Hemolytic Properties of Materials. West Conshohocken, PA, USA: ASTM International, 2000

[51] Zhang X, Li M, Wang Y B, . Cell response of nanographene platelets to human osteoblast-like MG63 cells. Journal of Biomedical Materials Research Part A, 2014, 102(3): 732–742

[52] La W G, Jung M J, Yoon J K, . Bone morphogenetic protein-2 for bone regeneration − Dose reduction through graphene oxide-based delivery. Carbon, 2014, 78: 428–438

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