Gene expression analyses of human macrophage phagocytizing sub-micro titanium particles by allergy DNA chip (Genopal)

Biomed Mater Eng. 2009;19(1):63-70. doi: 10.3233/BME-2009-0564.

Abstract

The purpose of this study was to examine gene expressions of macrophage phagocytizing sub-micro Ti particles by a DNA chip. Human monocytic cell line THP-1 was differentiated into macrophages by culturing for two days in medium supplemented with 200 nM phorbol ester (PMA). The macrophages were then cultured in four media: medium without PMA (control); medium with suspended sub-micro Ti particles (0.5 wt%); medium with 1.0 microg/ml lipopolysaccharide (LPS); and medium with LPS and Ti particles. After 6 hours' culture, total RNA were extracted and gene expressions were evaluated by DNA allergy chip with 205 allergy and inflammation related gene spots. We found that phagocytosis of sub-micro Ti particles and LPS independently and synergistically up-regulated 17 inflammation-related genes more than two-fold. The extensive expressions of four genes (CCL1, IL1B, IL6 and IL8) were further confirmed by real-time quantitative PCR. It turned out that dual stimulation of LPS and Ti particles most up-regulated three genes (IL1B, IL6 and IL8), followed by LPS while Ti particles moderately but least increased, suggesting that phagocytosis of sub-micro Ti particles induces moderate inflammation with its degree less than LPS, but phagocytosis of sub-micro Ti particles has the potential to worsen inflammation caused by LPS-stimulated macrophages.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay / methods*
  • Cells, Cultured
  • Cytokines / analysis
  • Cytokines / metabolism*
  • Gene Expression Profiling / methods*
  • Humans
  • Macrophages / physiology*
  • Nanoparticles*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Phagocytosis / physiology*
  • Titanium*

Substances

  • Cytokines
  • Titanium