Single-walled carbon nanotubes functionalized with sodium hyaluronate enhance bone mineralization

Braz J Med Biol Res. 2016 Feb;49(2):e4888. doi: 10.1590/1414-431X20154888. Epub 2015 Dec 4.


The aim of this study was to evaluate the effects of sodium hyaluronate (HY), single-walled carbon nanotubes (SWCNTs) and HY-functionalized SWCNTs (HY-SWCNTs) on the behavior of primary osteoblasts, as well as to investigate the deposition of inorganic crystals on titanium surfaces coated with these biocomposites. Primary osteoblasts were obtained from the calvarial bones of male newborn Wistar rats (5 rats for each cell extraction). We assessed cell viability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay and by double-staining with propidium iodide and Hoechst. We also assessed the formation of mineralized bone nodules by von Kossa staining, the mRNA expression of bone repair proteins, and the deposition of inorganic crystals on titanium surfaces coated with HY, SWCNTs, or HY-SWCNTs. The results showed that treatment with these biocomposites did not alter the viability of primary osteoblasts. Furthermore, deposition of mineralized bone nodules was significantly increased by cells treated with HY and HY-SWCNTs. This can be partly explained by an increase in the mRNA expression of type I and III collagen, osteocalcin, and bone morphogenetic proteins 2 and 4. Additionally, the titanium surface treated with HY-SWCNTs showed a significant increase in the deposition of inorganic crystals. Thus, our data indicate that HY, SWCNTs, and HY-SWCNTs are potentially useful for the development of new strategies for bone tissue engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Bone Morphogenetic Protein 2 / metabolism
  • Bone Morphogenetic Protein 4 / metabolism
  • Calcification, Physiologic / drug effects*
  • Cell Survival
  • Coated Materials, Biocompatible / pharmacology
  • Collagen Type I / metabolism
  • Collagen Type III / metabolism
  • Hyaluronic Acid / pharmacology*
  • Male
  • Microscopy, Electron, Scanning
  • Nanotubes, Carbon* / chemistry
  • Organometallic Compounds / pharmacology
  • Osteoblasts / drug effects*
  • Primary Cell Culture
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • Rats, Wistar
  • Spectrometry, X-Ray Emission
  • Staining and Labeling / methods
  • Tissue Engineering / methods
  • Titanium / chemistry
  • Titanium / metabolism*


  • Bmp2 protein, rat
  • Bmp4 protein, rat
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Protein 4
  • Coated Materials, Biocompatible
  • Collagen Type I
  • Collagen Type III
  • Nanotubes, Carbon
  • Organometallic Compounds
  • RNA, Messenger
  • Hyaluronic Acid
  • Titanium