In this study, a three-phased multiwalled scaffold, composed of carbon nanotube (mwCNT), nanocrystalline hydroxyapatite (nHA), and polycaprolactone (PCL), was fabricated by the solvent evaporation technique. The structure character, mechanical properties, and degradation activity in simulated body fluid (SBF), along with osteoproductive ability in human osteosarcoma cell MG63, were investigated thoroughly. Results showed that the three phases in mwCNT/nHA/PCL composite presented excellent miscibility and stronger interfacial force when the weight content was 1/15/84 (wt%). Simultaneously, the composite had smaller porosity and slower degradation rate, and there was massive crystallized hydroxyapatite formed on the surface after being soaked in SBF. With regard to bioactivity, MG63s on this scaffolds presented good proliferation performance and differentiated into the osteogenic lineage by expressing high levels of ALP. It was concluded that mwCNTs/nHA/PCL composite scaffolds might be beneficial for bone tissue engineering at a relatively low concentration of mwCNTs and nHA.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
U. Kneser, D.J. Schaefer, B. Munder, C. Klemt, C. Andree, and G.B. Stark: Tissue engineering of bone. Minimally Invasive Ther. Allied Technol. 11, 107 (2004).
D. Dean, K.J. Min, and A. Bond: Computer aided design of large-format prefabricated cranial plates. J. Craniofacial Surg. 14, 819 (2003).
T.L. Siu, J.M. Rogers, K. Lin, R. Thompson, and M. Owbridge: Custom-made titanium 3D printed interbody cages for treatment of osteoporotic fracture related spinal deformity. World Neurosurg. 111, 1 (2018).
D.W. Hutmacher: Scaffolds in tissue engineering bone and cartilage. Biomaterials 21, 2529 (2000).
S.J. Hollister: Porous scaffold design for tissue engineering. Nat. Mater. 4, 518 (2005).
I. Armentano, M. Dottori, E. Fortunati, S. Mattioli, and J.M. Kenny: Biodegradable polymer matrix nanocomposites for tissue engineering: A review. Polym. Degrad. Stab. 95, 2126 (2010).
M. Abu Bakar, P. Cheang, and K. Khor: Mechanical properties of injection molded hydroxyapatite-polyetheretherketone biocomposites. Compos. Sci. Technol. 63, 421 (2003).
K. Li and S.C. Tjong: Preparation and characterization of isotactic polypropylene reinforced with hydroxyapatite nanorods. J. Macromol. Sci., Phys. 50, 1983 (2011).
D. Tasis, N. Tagmatarchis, A. Bianco, and M. Prato: Chemistry of carbon nanotubes. Chem. Rev. 106, 1105 (2006).
K. Sahithi, M. Swetha, K. Ramasamy, N. Srinivasan, and N. Selvamurugan: Polymeric composites containing carbon nanotubes for bone tissue engineering. Int. J. Biol. Macromol. 46, 281 (2010).
Q. Cheng, K. Rutledge, and E. Jabbarzadeh: Carbon nanotube—poly(lactide-co-glycolide) composite scaffolds for bone tissue engineering applications. Ann. Biomed. Eng. 41, 904 (2013).
C.Z. Liao, K. Li, H.M. Wong, W.Y. Tong, K.W.K. Yeung, and S.C. Tjong: Novel polypropylene biocomposites reinforced with carbon nanotubes and hydroxyapatite nanorods for bone replacements. Mater. Sci. Eng., C 33, 1380 (2013).
T. Shokuhfar, A. Makradi, E. Titus, G. Cabral, S. Ahzi, A.C. Sousa, S. Belouettar, and J. Gracio: Prediction of the mechanical properties of hydroxyapatite/polymethyl methacrylate/carbon nanotubes nanocomposite. J. Nanosci. Nanotechnol. 8, 4279 (2008).
J. Zhang, Z. Wen, M. Zhao, and C. Dai: Effect of the addition CNTs on performance of CaP/chitosan/coating deposited on magnesium alloy by electrophoretic deposition. Mater. Sci. Eng., C 58, 992 (2016).
S. Liao, G.F. Xu, W. Wang, F. Watari, F.Z. Cui, S. Ramakrishna, and C.K. Chan: Self-assembly of nano-hydroxyapatite on multi-walled carbon nanotubes. Acta Biomater. 3, 669 (2007).
M.A. Woodruff and D.W. Hutmacher: The return of a forgotten polymer—polycaprolactone in the 21st century. Prog. Polym. Sci. 35, 1217 (2010).
J. Huang, S.M. Best, W. Bonfield, R.A. Brooks, N. Rushton, S.N. Jayasinghe, and M.J. Edirisinghe: In vitro assessment of the biological response to nano-sized hydroxyapatite. J. Mater. Sci.: Mater. Med. 15, 441 (2004).
B. Dorj, J.E. Won, J.H. Kim, S.J. Choi, U.S. Shin, and H.W. Kim: Robocasting nanocomposite scaffolds of poly(caprolactone)/hydroxyapatite incorporating modified carbon nanotubes for hard tissue reconstruction. J. Biomed. Mater. Res., Part A 101, 1670 (2013).
E.M. Goncalves, F.J. Oliveira, R.F. Silva, M.A. Neto, M.H. Fernandes, M. Amaral, M.V. Regí, and M. Vila: Three-dimensional printed PCL-hydroxyapatite scaffolds filled with CNTs for bone cell growth stimulation. J. Biomed. Mater. Res., Part B 104, 1210 (2015).
A. Baji, S.C. Wong, T. Liu, T. Li, and T.S. Srivatsan: Morphological and X-ray diffraction studies of crystalline hydroxyapatite-reinforced polycaprolactone. J. Biomed. Mater. Res., Part B 81, 343 (2007).
H.W. Kim, J.C. Knowles, and H.E. Kim: Hydroxyapatite/poly(ε-caprolactone) composite coatings on hydroxyapatite porous bone scaffold for drug delivery. Biomaterials 25, 1279 (2004).
M. Cadek, J.N. Coleman, V. Barron, K. Hedicke, and W.J. Blau: Morphological and mechanical properties of carbon-nanotube-reinforced semicrystalline and amorphous polymer composites. Appl. Phys. Lett. 81, 5123 (2003).
T. Kokubo, H.M. Kim, and M. Kawashita: Novel bioactive materials with different mechanical properties. Biomaterials 24, 2161 (2003).
D. Boyd, M.R. Towler, A.W. Wren, O.M. Clarkin, and D.A. Tanner: TEM analysis of apatite surface layers observed on zinc based glass polyalkenoate cements. J. Mater. Sci. 43, 1170 (2008).
X. Zhong, Z.F. Lu, P. Valtchev, H. Wei, H. Zreiqat, and F. Dehghani: Surface modification of poly(propylene carbonate) by aminolysis and layer-by-layer assembly for enhanced cytocompatibility. Colloids Surf., B 93, 75 (2012).
Y.S. Hwang, N. Sangaj, and S. Varghese: Interconnected macroporous poly(ethylene glycol) cryogels as a cell scaffold for cartilage tissue engineering. Tissue Eng., Part A 16, 3033 (2010).
K. Anselme, P. Linez, M. Bigerelle, D. Le Maguer, P. Hardouin, H.F. Hildebrand, A. Iost, and J.M. Leroy: The relative influence of the topography and chemistry of TiAl6V4 surfaces on osteoblastic cell behaviour. Biomaterials 21, 1567 (2000).
B. Tucker and M. Lardelli: A rapid apoptosis assay measuring relative acridine orange fluorescence in zebrafish embryos. Zebrafish 4, 113 (2007).
E. Vega-Avila and M.K. Pugsley: An overview of colorimetric assay methods used to assess survival or proliferation of mammalian cells. Proc. West. Pharmacol. Soc. 54, 10 (2011).
K. Anselme: Osteoblast adhesion on biomaterials. Biomaterials 21, 667 (2000).
V. Neuhoff, R. Stamm, and H. Eibl: Clear background and highly sensitive protein staining with Coomassie Blue dyes in polyacrylamide gels: A systematic analysis. Electrophoresis 6, 427 (1985).
C.Y. Li and T.W. Chou: Modeling of elastic buckling of carbon nanotubes by molecular structural mechanics approach. Mech. Mater. 36, 1047 (2004).
Y.Z. Zhang, J.R. Venugopal, A. El-Turki, S. Ramakrishna, B. Su, and C.T. Lim: Electrospun biomimetic nanocomposite nanofibers of hydroxyapatite/chitosan for bone tissue engineering. Biomaterials 29, 4314 (2008).
S.Z. Fu, P.Y. Ni, B.Y. Wang, B.Y. Chu, J.R. Peng, L. Zheng, X. Zhao, F. Luo, Y.Q. Wei, and Z.Y. Qian: In vivo biocompatibility and osteogenesis of electrospun poly(ε-caprolactone)–poly(ethylene glycol)–poly(ε-caprolactone)/nano-hydroxyapatite composite scaffold. Biomaterials 33, 8363 (2012).
Y.B. Zhang, V. Leblanc-Boily, Y. Zhao, and R.E. Prud’homme: Wide angle X-ray diffraction investigation of crystal orientation in miscible blend of poly(ε-caprolactone)/poly(vinyl chloride) crystallized under strain. Polymer 46, 8141 (2005).
K. Cho, D.N. Saheb, H. Yang, B. Kang, J. Kim, and S. Lee: Real time in situ X-ray diffraction studies on the melting memory effect in the crystallization of β-isotactic polypropylene. Polymer 44, 4053 (2003).
B. McCarthy, J.N. Coleman, R. Czerw, A.B. Dalton, M.I.H. Panhuis, A. Maiti, A. Drury, P. Bernier, J.B. Nagy, B. Lahr, H.J. Byrne, D.L. Carroll, and W.J. Blau: A microscopic and spectroscopic study of interactions between carbon nanotubes and a conjugated polymer. J. Phys. Chem. B 106, 2210 (2002).
L.L. Pan, X.B. Pei, R. He, Q.B. Wan, and J. Wang: Multiwall carbon nanotubes/polycaprolactone composites for bone tissue engineering application. Colloids Surf., B 93, 226 (2012).
F. Luo, L.L. Pan, G. Hong, T. Wang, X.B. Pei, J. Wang, and Q.B. Wan: In vitro and in vivo characterization of multi-walled carbon nanotubes/polycaprolactone composite scaffolds for bone tissue engineering applications. J. Biomater. Tissue Eng. 7, 787 (2017).
J.E. Babensee, J.M. Anderson, L.V. McIntire, and A.G. Mikos: Host response to tissue engineered devices. Adv. Drug Delivery Rev. 33, 111 (1998).
O. Persenaire, M. Alexandre, P. Degee, and P. Dubois: Mechanisms and kinetics of thermal degradation of poly(ε-caprolactone). Biomacromolecules 2, 288 (2001).
C.X.F. Lam, M.M. Savalani, S.H. Teoh, and D.W. Hutmacher: Dynamics of in vitro polymer degradation of polycaprolactone-based scaffolds: Accelerated versus simulated physiological conditions. Biomed. Mater. 3, 4108 (2008).
J.M. Meseguer-Duenas, J. Mas-Estelles, I. Castilla-Cortazar, J.L. Escobar Ivirico, and A. Vidaurre: Alkaline degradation study of linear and network poly(ε-caprolactone). J. Mater. Sci.: Mater. Med. 22, 11 (2011).
Y. Lei, B. Rai, K.H. Ho, and S.H. Teoh: In vitro degradation of novel bioactive polycaprolactone—20% tricalcium phosphate composite scaffolds for bone engineering. Mater. Sci. Eng., C 27, 293 (2007).
M. Cadek, J.N. Coleman, V. Barron, K. Hedicke, and W.J. Blau: Morphological and mechanical properties of carbon-nanotube-reinforced semicrystalline and amorphous polymer composites. Appl. Phys. Lett. 81, 5123 (2002).
T. Kokubo and H. Takadama: How useful is SBF in predicting in vivo bone bioactivity? Biomaterials 27, 2907 (2006).
G. Thalji, C. Gretzer, and L.F. Cooper: Comparative molecular assessment of early osseointegration in implant-adherent cells. Bone 52, 444 (2013).
D.O. Costa, P.D.H. Prowse, T. Chrones, S.M. Sims, D.W. Hamilton, A.S. Rizkalla, and S. Jeffrey Dixon: The differential regulation of osteoblast and osteoclast activity by surface topography of hydroxyapatite coatings. Biomaterials 34, 7215 (2013).
L.E. Freed, G. Vunjak-Novakovic, R.J. Biron, D.B. Eagles, D.C. Lesnoy, S.K. Barlow, and R. Langer: Biodegradable polymer scaffolds for tissue engineering. Biotechnology 12, 689 (1994).
D. Yamashita, M. Machigashira, M. Miyamoto, H. Takeuchi, K. Noguchi, Y. Izumi, and S. Ban: Effect of surface roughness on initial responses of osteoblast-like cells on two types of zirconia. Dent. Mater. J. 28, 461 (2009).
D. Mata, F.J. Oliveira, M. Ferro, P.S. Gomes, M.H. Fernandes, M.A. Lopes, and R.F. Silva: Multifunctional carbon nanotube/bioceramics modulate the directional growth and activity of osteoblastic cells. J. Biomed. Nanotechnol. 10, 725 (2014).
J. Behring, R. Junker, X.F. Walboomers, B. Chessnut, and J.A. Jansen: Toward guided tissue and bone regeneration: Morphology, attachment, proliferation, and migration of cells cultured on collagen barrier membranes. A systematic review. Odontology 96, 1 (2008).
G. Raffaini and F. Ganazzoli: Surface topography effects in protein adsorption on nanostructured carbon allotropes. Langmuir 29, 4883 (2013).
H.U. Lee, Y.S. Jeong, S.Y. Jeong, S.Y. Park, J.S. Bae, H.G. Kim, and C.R. Cho: Role of reactive gas in atmospheric plasma for cell attachment and proliferation on biocompatible poly ε-caprolactone film. Appl. Surf. Sci. 254, 5700 (2008).
R. Marom, I. Shur, R. Solomon, and D. Benayahu: Characterization of adhesion and differentiation markers of osteogenic marrow stromal cells. J. Cell. Physiol. 202, 41 (2005).
S. Morelli, S. Salerno, J. Holopainen, M. Ritala, and L.D. Bartolo: Osteogenic and osteoclastogenic differentiation of co-cultured cells in polylactic acid—nanohydroxyapatite fiber scaffolds. J. Biotechnol. 204, 53 (2015).
A. Abarrategi, M.C. Gutierrez, C. Moreno-Vicente, M.J. Hortiguela, V. Ramos, J.L. Lopez-Lacomba, M.L. Ferrer, and F.D. Monte: Multiwall carbon nanotube scaffolds for tissue engineering purposes. Biomaterials 29, 94 (2008).
O. Im, J. Li, M. Wang, L.G. Zhang, and M. Keidar: Biomimetic three-dimensional nanocrystalline hydroxyapatite and magnetically synthesized single-walled carbon nanotube chitosan nanocomposite for bone regeneration. Int. J. Nanomed. 7, 2087 (2012).
About this article
Cite this article
Yang, H., Li, J., Liao, Q. et al. In vitro evaluation of a novel multiwalled carbon nanotube/nanohydroxyapatite/polycaprolactone composite for bone tissue engineering. Journal of Materials Research 34, 532–544 (2019). https://doi.org/10.1557/jmr.2018.484