Titanium particles stimulate COX-2 expression in synovial fibroblasts through an oxidative stress-induced, calpain-dependent, NF-kappaB pathway

Am J Physiol Cell Physiol. 2009 Aug;297(2):C310-20. doi: 10.1152/ajpcell.00597.2008. Epub 2009 Jun 3.

Abstract

In prosthetic loosening, bone resorption is induced by wear debris particles generated from the artificial joint articulation. Our prior work showed that synovial-like fibroblasts respond to titanium particles by producing receptor activator of NF-kappaB ligand (RANKL), a critical activator of osteoclastogenesis. While this effect occurs through a cyclooxygenase-2 (COX-2)-dependent pathway, the mechanism of COX-2 stimulation by titanium particles is not clear. Here we show that titanium particles induce COX-2 gene expression by activating NF-kappaB signaling. Inhibitor of NF-kappaB (IkappaBalpha) is degraded following particle treatment, permitting active NF-kappaB to translocate to the nucleus where it interacts with the COX-2 promoter and drives transcription. NF-kappaB activation is dependent on reactive oxygen species since antioxidants block the NF-kappaB signaling induced by particles. Surprisingly, IkappaBalpha degradation is independent of IKK (IkappaB kinase) and the 26S proteasome. Instead, calpain inhibitor can block the IkappaBalpha degradation induced by particles. Furthermore, the calpain-targeted COOH-terminal PEST sequence of IkappaBalpha is necessary for phosphorylation and degradation, consistent with a proteasome-independent mechanism of catabolism. Altogether, the data demonstrate a signaling pathway by which titanium particles induce oxidative stress, stimulate calpain-mediated NF-kappaB activation, and activate target gene expression, including COX-2. These findings define important targets for osteolysis but may also have importance in other diseases where fibroblasts respond to environmental particles, including pulmonary diseases.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Calpain / genetics
  • Calpain / metabolism*
  • Cells, Cultured
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 / metabolism*
  • Fibroblasts / cytology
  • Fibroblasts / physiology*
  • Gene Expression Regulation
  • Humans
  • I-kappa B Proteins / metabolism
  • Mice
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Oxidative Stress*
  • Promoter Regions, Genetic
  • RANK Ligand / genetics
  • RANK Ligand / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction / physiology
  • Synovial Membrane / cytology*
  • Titanium / metabolism*

Substances

  • I-kappa B Proteins
  • NF-kappa B
  • NFKBIA protein, human
  • Nfkbia protein, mouse
  • RANK Ligand
  • Recombinant Fusion Proteins
  • NF-KappaB Inhibitor alpha
  • Titanium
  • Cyclooxygenase 2
  • Calpain