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Carbon Nanotube Reinforced Collagen/Hydroxyapatite Scaffolds Improve Bone Tissue Formation In Vitro and In Vivo

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Abstract

Current bone regeneration strategies faced major challenges in fabricating the bionic scaffolds with nano-structure, constituents and mechanical features of native bone. In this study, we developed a new porous scaffold by adding the multi-walled carbon nanotube (MWCNT) into collagen (Col)/hydroxyapatite (HA) composites. Data showed that 0.5%CNT/Col/HA (0.5%CNT) group was approximately tenfolds stiffer than Col–HA, and it was superior in promoting bone marrow mesenchymal stem proliferation and spreading, mRNA and protein expressions of bone sialoprotein (BSP) and osteocalcin (OCN) than Col–HA group. Moreover, we utilized 0.5%CNT composite to repair the rat calvarial defects (8 mm diameter) in vivo, and observed the new bone formation by 3D reconstruction of micro CT, HE and Masson staining, and BSP, OCN by immunohistochemical analysis. Results showed that newly formed bone in 0.5%CNT group was significantly higher than that in Col–HA group at 12 weeks. These findings highlighted a promising strategy in healing of large area bone defect with MWCNT added into the Col–HA scaffold as they possessed the combined effects of mechanical strength and osteogenicity.

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References

  1. Bettinger, C. J., R. Langer, and J. T. Borenstein. Engineering substrate topography at the micro- and nanoscale to control cell function. Angew. Chem. Int. Ed. Engl. 48:5406–5415, 2009.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Chen, X., U. C. Tam, and J. L. Czlapinski. Interfacing carbon nanotubes with living cells. J. Am. Chem. Soc. 128:6292–6293, 2006.

    Article  CAS  PubMed  Google Scholar 

  3. Cheng, Q., K. Rutledge, and E. Jabbarzadeh. Carbon nanotube-poly(lactide-co-glycolide) composite scaffolds for bone tissue engineering applications. Ann. Biomed. Eng. 41:904–916, 2013.

    Article  PubMed  Google Scholar 

  4. Christenson, R. H. Biochemical markers of bone metabolism: an overview. Clin. Biochem. 30:573–593, 1997.

    Article  CAS  PubMed  Google Scholar 

  5. Cicciù, M., A. S. Herford, G. Juodžbalys, and E. Stoffella. Recombinant human bone morphogenetic protein type 2 application for a possible treatment of bisphosphonates-related osteonecrosis of the jaw. J. Craniofac. Surg. 23:784–788, 2012.

    Article  PubMed  Google Scholar 

  6. Cranford, S. W., J. Boer, C. van Blitterswijk, and M. J. Buehler. Materiomics: an -omics approach to biomaterials research. Adv. Mater., 25:802−824, 2013.

    Article  CAS  PubMed  Google Scholar 

  7. Ganss, B., R. H. Kim, and J. Sodek. Bone sialoprotein. Crit. Rev. Oral Biol. Med. 10:79–98, 1999.

    Article  CAS  PubMed  Google Scholar 

  8. Harrison, B. S., and A. Atala. Carbon nanotube applications for tissue engineering. Biomaterials 28:344–353, 2007.

    Article  CAS  PubMed  Google Scholar 

  9. Herford, A. S., and M. Cicciù. Recombinant human bone morphogenetic protein type 2 jaw reconstruction in patients affected by giant cell tumor. J. Craniofac. Surg. 21:1970–1975, 2010.

    Article  PubMed  Google Scholar 

  10. Herford, A. S., M. Cicciù, and L. F. Eftimie. rhBMP-2 applied as support of distraction osteogenesis: A split-mouth histological study over nonhuman primates mandibles. Int. J. Clin. Exp. Med. 9:17187–17194, 2016.

    Google Scholar 

  11. Herford, A. S., R. Tandon, T. W. Stevens, E. Stoffella, and M. Cicciu. Immediate distraction osteogenesis: the sandwich technique in combination with rhBMP-2 for anterior maxillary and mandibular defects. J. Craniofac. Surg. 24:1383–1387, 2013.

    Article  PubMed  Google Scholar 

  12. Laino, L., I. Giovanna, P. Adriano, L. M. Lorenzo, and M. Cicciù. Vertical ridge augmentation of the atrophic posterior mandible with sandwich technique: bone block from the chin area versus corticocancellous bone block allograft–clinical and histological prospective randomized controlled study. Biomed. Res. Int. 2014:982104, 2014.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Liao, S. S., F. Z. Cui, W. Zhang, and Q. L. Feng. Hierarchically biomimetic bone scaffold materials: nano-HA/collagen/PLA composite. J. Biomed. Mater. Res. B Appl. Biomater. 69:158–165, 2004.

    Article  CAS  PubMed  Google Scholar 

  14. Marco, C., A. S. Herford, D. Cicciù, R. Tandon, and C. Maiorana. Recombinant human bone morphogenetic protein-2 promote and stabilize hard and soft tissue healing for large mandibular new bone reconstruction defects. J. Craniofac. Surg. 25:860–862, 2014.

    Article  Google Scholar 

  15. Namgung, S., K. Y. Baik, J. Park, and S. Hong. Controlling the growth and differentiation of human mesenchymal stem cells by the arrangement of individual carbon nanotubes. ACS Nano. 5:7383–7390, 2011.

    Article  CAS  PubMed  Google Scholar 

  16. Narita, N., Y. Kobayashi, and H. Nakamura. Multiwalled carbon nanotubes specifically inhibit osteoclast differentiation and function. Nano Lett. 9:1406–1413, 2009.

    Article  CAS  PubMed  Google Scholar 

  17. Nel, A. E., L. Mädler, D. Velegol, T. Xia, E. M. Hoek, P. Somasundaran, F. Klaessig, V. Castranova, and M. Thompson. Understanding biophysicochemical interactions at the nano-bio interface. Nat. Mater. 8:543–557, 2009.

    Article  CAS  PubMed  Google Scholar 

  18. Newman, P., A. Minett, R. Ellis-Behnke, and H. Zreiqat. Carbon nanotubes: their potential and pitfalls for bone tissue regeneration and engineering. Nanomedicine. 9:1139–1158, 2013.

    Article  CAS  PubMed  Google Scholar 

  19. Niu, L., H. Kua, and D. H. Chua. Bonelike apatite formation utilizing carbon nanotubes as template. Langmuir 26:4069–4073, 2010.

    Article  CAS  PubMed  Google Scholar 

  20. Oh, S., K. S. Brammer, Y. S. J. Li, D. Teng, A. J. Engler, and S. Chien. Stem cell fate dictated solely by altered nanotube dimension. Proc. Natl. Acad. Sci. USA 106:2130–2135, 2009.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Petrauskaite, O., P. Gomes, and M. H. Fernandes. Biomimetic mineralization on a microporous cellulose-based matrix for bone regeneration. Biomed. Res. Int. 2013:452750, 2013.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Shen, X., L. Chen, X. Cai, T. Tong, H. Tong, and J. Hu. A novel method for the fabrication of homogeneous hydroxyapatite/collagen nanocomposite and nanocomposite scaffold with hierarchical porosity. J. Mater. Sci. Mater. Med. 22:299–305, 2011.

    Article  CAS  PubMed  Google Scholar 

  23. Shimizu, M., Y. Kobayashi, and T. Mizoguchi. Carbon nanotubes induce bone calcification by bidirectional interaction with osteoblasts. Adv. Mater. 24:2176–2185, 2012.

    Article  CAS  PubMed  Google Scholar 

  24. Wang, S. F., L. Shen, W. D. Zhang, and Y. J. Tong. Preparation and mechanical properties of chitosan/carbon nanotubes composites. Biomacromol 6:3067–3072, 2005.

    Article  CAS  Google Scholar 

  25. Wick, P., P. Manser, and L. K. Limbach. The degree and kind of agglomeration affect carbon nanotube cytotoxicity. Toxicol. Lett. 168:121–131, 2007.

    Article  CAS  PubMed  Google Scholar 

  26. Xiao, Y., T. Gong, and S. Zhou. The functionalization of multi-walled carbon nanotubes by in situ deposition of hydroxyapatite. Biomaterials 31:5182–5190, 2010.

    Article  CAS  PubMed  Google Scholar 

  27. Zanello, L. P., B. Zhao, H. Hu, and R. C. Haddon. Bone cell proliferation on carbon nanotubes. Nano Lett. 6:562–567, 2006.

    Article  CAS  PubMed  Google Scholar 

  28. Zhao, B., H. Hu, S. K. Mandal, and R. C. Haddon. A bone mimic based on the self-assembly of hydroxyapatite on chemically functionalized single-walled carbon nanotubes. Chem. Mater. 17:3235–3241, 2005.

    Article  CAS  Google Scholar 

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Acknowledgment

This work was supported by grants from National Nature Science Foundation of China (Nos. 31271052, 31470904, 31670992), Sichuan Provincial Science and Technology Department Fund (No. 2013SZ0057), and Medical Scientific Research Projects of Chongqing Health Department (No. 2013-2-070), Program for Innovation Team Building at Institutions of Higher Education in Chongqing in 2016 (CXTDG201602006), and Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education.

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Authors and Affiliations

  1. Chongqing Key Laboratory of Oral Diseases and Biomedical Science, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Stomatological Hospital of Chongqing Medical University, 426#, Songshi North Road, Chongqing, 401147, People’s Republic of China

    Zheng Jing & Li Cao

  2. Department of Stomatology, Affiliated Hospital of Chengdu University of TCM, No. 39 Shierqiao Road, Chengdu, 610075, People’s Republic of China

    Yeke Wu

  3. National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, 610064, People’s Republic of China

    Wen Su

  4. State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, China 14#, Section 3rd, South Renmin Rd, Chengdu, 610041, People’s Republic of China

    Mi Tian, Wenlu Jiang, Lixing Zhao & Zhihe Zhao

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  1. Zheng Jing
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  2. Yeke Wu
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  3. Wen Su
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  4. Mi Tian
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  6. Li Cao
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  7. Lixing Zhao
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  8. Zhihe Zhao
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Correspondence to Lixing Zhao.

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Associate Editor Debra T. Auguste oversaw the review of this article.

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Jing, Z., Wu, Y., Su, W. et al. Carbon Nanotube Reinforced Collagen/Hydroxyapatite Scaffolds Improve Bone Tissue Formation In Vitro and In Vivo . Ann Biomed Eng 45, 2075–2087 (2017). https://doi.org/10.1007/s10439-017-1866-9

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  • DOI: https://doi.org/10.1007/s10439-017-1866-9

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