Differentiation of mesenchymal stem cells onto highly adherent radio frequency-sputtered carbonated hydroxylapatite thin films

J Biomed Mater Res A. 2010 Dec 15;95(4):1203-14. doi: 10.1002/jbm.a.32947. Epub 2010 Oct 11.

Abstract

In this work, an improved version of the radio frequency magnetron sputtering (RF-MS) technique was used to prepare highly adherent B-type carbonated hydroxylapatite (B-CHA) thin films. Fourier transform infrared spectroscopy (FTIR) and grazing incidence X-ray diffraction studies proved that the coatings maintained the composition and revealed the polycrystalline structure of HA. Scanning electron microscopy analysis showed that the CHA films are rough and exhibit a homogeneous microstructure. Energy-dispersive X-ray spectroscopy (EDX) mapping demonstrated a uniform distribution of the Ca and P cations while a Ca/P ratio of 1.8 was found. In addition, the FTIR experiments showed a remarkable reproducibility of the nanostructures. Human mesenchymal stem cells (hMSCs), in vitro differentiated osteoblasts, and explanted bone cells were grown over the surface of CHA coatings for periods between a few hours and 21 days. Osteoprogenitor cells maintained viability and characteristic morphology after adhesion on CHA coatings. Ki67-positive osteoblasts were the evidence of cell proliferation events. Cells showed positive staining for markers of osteoblast phenotype such as collagen type I, bone sialoprotein and osteonectin. Our data showed the formation of mineralized foci by differentiation of hMSCs to human primary osteoblasts after cultivation in osteogenic media on RF-sputtered films. The results demonstrate the capacity of B-type CHA coating to support MSCs adhesion and osteogenic differentiation ability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism
  • Cell Adhesion / drug effects
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Cell Shape / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Fluorescent Antibody Technique
  • Humans
  • Hydroxyapatites / pharmacology*
  • Materials Testing / methods*
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / ultrastructure
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Osteoblasts / metabolism
  • Radio Waves*
  • Silicon Dioxide / pharmacology
  • Spectrometry, X-Ray Emission
  • Spectroscopy, Fourier Transform Infrared
  • Titanium / pharmacology
  • X-Ray Diffraction

Substances

  • Biomarkers
  • Hydroxyapatites
  • Silicon Dioxide
  • Titanium