Preparation and characterizations of three-dimensional porous collagen/graphene oxide/hydroxyapatite nanocomposite scaffolds for bone tissue engineering

Bhahat Lawlley Zimba, Jiang Hao, Chen Li, Li Yaomin, Yu Xunzhi, Chen Chang, Wan Jiangling, Qingzhi Wu


Studies have reported that the incorporation of graphene oxide (GO) and hydroxyapatite (HA) into biocompatible polymers (such as collagen (Col), chitosan, alginate, etc) results in enhanced structural and mechanical properties respectively. The objective of this study was to prepare and characterize three-dimensional (3D) porous Col/GO/HA nanocomposite scaffolds and to investigate cytocompatibility and osteogenic differentiation potential of rat bone marrow mesenchymal stem cells (rBMSCs) on the as-prepared scaffolds. The SEM images revealed that the scaffolds were porous with the pore diameter inversely proportional to the concentration of HA. XRD results were able to depict the characteristic peaks for HA which shows that HA was incorporated into the scaffolds. The rBMSCs which were cultured on the scaffolds were able to attach and proliferate during the 21 days of the experiment which indicates that the as-prepared scaffolds are cytocompatible. The Alizarin red staining demonstrated the presence of calcium deposits as there were orange-red stains on the samples after culturing the cells using the osteogenic differentiation medium. These results demonstrate the promising potential of the 3D porous Col/GO/HA nanocomposite scaffolds for applications in bone tissue engineering.


Collagen, Graphene oxide, Hydroxyapatite, Porous scaffolds, Tissue engineering

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Chen F., L.X., Advancing biomaterials of human origin for tissue engineering. Prog Polym Sci, 2016. 53: p. 86–168.

F.J., O.B., Biomaterials & scaffolds for tissue engineering. Materialstoday, 2011. 14(3): p. 88-95.

Sultana N., L.M.M., Composite Synthetic Scaffolds for Tissue Engineering and Regenerative Medicine. Springer Briefs in Materials, 2015.

Gopal S.K., U.K.S., Sowndarya S., Suresh, P.B., Rose, C., Isolation, characterization, and in vitro evaluation of bovine rumen submucosa films of collagen or chitosan-treated collagen. J Biomater Appl., 2016. 30(6): p. 780-792.

P., K., Laser flash synthesis of graphene and its inorganic analogues: an innovative breakthrough with immense promise. RSC Adv., 2013. 3: p. 11987–12002.

Nishida E., M.H., Kato A., et al., Graphene oxide scaffold accelerates cellular proliferative response and alveolar bone healing of tooth extraction socket. Int. J. Nanomed., 2016. 11: p. 2265–2277.

Girão A.F., G.G., Bhangra K.S., et al., Electrostatic self-assembled graphene oxide-collagen scaffolds towards a three-dimensional microenvironment for biomimetic applications. RSC Adv., 2016. 6: p. 49039-49051.

Wang A., P.K., Dong B., et al. , Role of surface charge and oxidative stress in cytotoxicity and genotoxicity of graphene oxide towards human lung fibroblast cells. J Appl Toxicol., 2013. 33(10): p. 1156-1164.

Zhong, Y., & Ji, B., Impact of cell shape on cell migration behavior on elastic substrate. Biofabrication, 2013. 5.

Thein-Han, W.W., & Xu, H.H.K., Collagen-Calcium Phosphate Cement Scaffolds Seeded with Umbilical Cord Stem Cells for Bone Tissue Engineering. TISSUE ENGINEERING, 2011. 17.

Al-Munajjed, A.A., & O’Brien, F.J. , Influence of a novel calcium-phosphate coating on the mechanical properties of highly porous collagen scaffolds for bone repair. Journal of the Mechanical Behavior of Biomedical Materials, 2009. 2: p. 138-146.

Chen D., Z.X., Chen S., et. al. , One-pot fabrication of FePt/reduced graphene oxide composites as highly active and stable electrocatalysts for the oxygen reduction reaction. Carbon, 2014. 68: p. 755–762.

Huang W., M.Z., Chen L., et. al., Synthesis and characterisation of fluorescent and biocompatible hydroxyapatite nanoparticles with cerium doping. Micro & Nano Letters, 2018. 13(5): p. 699–703.

Lee J.H., S.Y.C., Lee S., et. al., Enhanced Osteogenesis by Reduced Graphene Oxide/Hydroxyapatite Nanocomposites. Scientific Reports 2015. 5.

Smajilagić A., A.M., Redžić A., et. al. , Rat bone marrow stem cells isolation and culture as a bone formative experimental system. Bosn J Basic Med Sci 2013. 13(1): p. 27-30.

Lee J.H., S.Y.C., Jin O.S., et. al. , Reduced graphene oxide-coated hydroxyapatite composites stimulate spontaneous osteogenic differentiation of human mesenchymal stem cells. Nanoscale, 2015. 7(1-9).

Jia L., D.Z., Fan D., et. al. , Human-like collagen/nano-hydroxyapatite scaffolds for the culture of chondrocytes. Mater Sci Eng C, 2013. 33: p. 727–734.

Gibson, L.J., Ashby, M., & Harley, B.A., Cellular Materials in Nature and Medicine. . 2010, Cambridge, UK: Cambridge University Press.

Haugh M.G., M.C.M., O’Brien F.J., Novel freeze-drying methods to produce a range of collagen–glycosaminoglycan scaffolds with tailored mean pore sizes. Tissue Eng C 2010. 16: p. 887–894.

Xia Z., Y.X., Jiang X., et al. , Fabrication and characterization of biomimetic collagen–apatite scaffolds with tunable structures for bone tissue engineering. Acta Biomater., 2013.

Venugopal J.R., P.M.P., Mukherjee S., et. al., Biomaterial strategies for alleviation of myocardial infarction. J Royal Soc Interface 2012. 9: p. 1-19.

Norahan M.H., A.M., Ghahremanzadeh R., et. al., Electroactive graphene oxide-incorporated collagen assisting vascularization for cardiac tissue engineering. J Biomed Mater Res., 2018: p. 1–16.

Brundavanam R.K, P.G.E.J., Fawcett D., Modelling the Crystal Structure of a 30 nm Sized Particle based Hydroxyapatite Powder Synthesised under the Influence of Ultrasound Irradiation from X-ray powder Diffraction Data. American Journal of Materials Science 2013. 3(4): p. 84-90.

Guan, J., Yang, J., Dai, J., Qin, Y., Wang, Y., Guo, Y., Ke, Q., & Zhang, C. , Bioinspired nanostructured hydroxyapatite/collagen three-dimensional porous scaffolds for bone tissue engineering. RSC Advances, 2015. 5(46): p. 36175–36184.

Chaudhuri B., B.D., Moroni L., Pramanik K., Myoblast differentiation of human mesenchymal stem cells on graphene oxide and electrospun graphene oxide–polymer composite fibrous meshes: Importance of graphene oxide conductivity and dielectric constant on their biocompatibility. Biofabrication, 2015. 7.

Yoruc A.B.H., A.A.K., Synthesis of Hydroxyapatite/Collagen (HA/COL) Composite Powder Using a Novel Precipitation Technique. ACTA PHYSICA POLONICA A., 2015. 127(4): p. 1264-1267.

Deepachitra R., R.V.S.T., Graphene oxide incorporated collagen–fibrin biofilm as a wound dressing material. RSC Adv., 2014. 4: p. 62717–62727.

Wahl D.A., C.J.T., Collagen-hydroxyapatite composites for hard tissue repair. European Cells and Materials 2006. 11: p. 43-56.

Chong Y., M.Y., Shen H., et. al. , The in vitro and in vivo toxicity of graphene quantum dots. Biomaterials, 2014. 35.

Park E. J., L.G.H., Han B. S., et. al., Toxic response of graphene nanoplatelets in vivo and in vitro. Arch. Toxicol., 2015. 89.

Li D., N.W., Chen L., et. al. , Self-Assembled Hydroxyapatite-Graphene Scaffold for Photothermal Cancer Therapy and Bone Regeneration. J. Biomed. Nanotechnol., 2018. 14: p. 2003-2017.

Wang L., Z.G., Liu H., et. al. , Nano-hydroxyapatite particles induce apoptosis on MC3T3-E1 cells and tissue cells in SD rats. Nanoscale, 2012. 4: p. 2894–2899

Tatavarty R., D.H., Lu G., Taylor R. J. & Bi X., Synergistic acceleration in the osteogenesis of human mesenchymal stem cells by graphene oxide–calcium phosphate nanocomposites. Chem. Commun., 2014. 50(62): p. 8484-8487.



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