Fetuin-A-containing calciprotein particles reduce mineral stress in the macrophage

PLoS One. 2013;8(4):e60904. doi: 10.1371/journal.pone.0060904. Epub 2013 Apr 8.

Abstract

The formation of fetuin-A-containing calciprotein particles (CPP) may facilitate the clearance of calcium phosphate nanocrystals from the extracellular fluid. These crystals may otherwise seed extra-osseous mineralization. Fetuin-A is a partially phosphorylated glycoprotein that plays a critical role in stabilizing these particles, inhibiting crystal growth and aggregation. CPP removal is thought to be predominantly mediated by cells of the reticuloendothelial system via type I and type II class A scavenger receptor (SR-AI/II). Naked calcium phosphate crystals are known to stimulate macrophages and other cell types in vitro, but little is known of the effect of CPP on these cells. We report here, for the first time, that CPP induce expression and secretion of tumour necrosis factor (TNF)-α, interleukin (IL)-1β in murine RAW 264.7 macrophages. Importantly, however, CPP induced significantly lower cytokine secretion than hydroxyapatite (HAP) crystals of equivalent size and calcium content. Furthermore, CPP only had a modest effect on macrophage viability and apoptosis, even at very high levels, compared to HAP crystals, which were strongly pro-apoptotic at much lower levels. Fetuin-A phosphorylation was found to modulate the effect of CPP on cytokine secretion and apoptosis, but not uptake via SR-AI/II. Prolonged exposure of macrophages to CPP was found to result in up-regulated expression of SR-AI/II. CPP formation may help protect against some of the pro-inflammatory and harmful effects of calcium phosphate nanocrystals, perhaps representing a natural defense system for calcium mineral stress. However, in pathological states where production exceeds clearance capacity, these particles may still stimulate pro-inflammatory and pro-apoptotic cascades in macrophages, which may be important in the pathogenesis of vascular calcification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Cytokines / metabolism
  • Durapatite / chemistry
  • Durapatite / pharmacology
  • Gene Expression Regulation / drug effects
  • Humans
  • Macrophages / cytology
  • Macrophages / drug effects*
  • Macrophages / metabolism*
  • Mice
  • Minerals / metabolism*
  • Oxidative Stress / drug effects
  • Phosphorylation / drug effects
  • Protein Transport / drug effects
  • Scavenger Receptors, Class A / genetics
  • Uremia / blood
  • alpha-2-HS-Glycoprotein / chemistry*
  • alpha-2-HS-Glycoprotein / metabolism
  • alpha-2-HS-Glycoprotein / pharmacology*

Substances

  • Cytokines
  • Minerals
  • Scavenger Receptors, Class A
  • alpha-2-HS-Glycoprotein
  • Durapatite

Grants and funding

This work was kindly supported by funding from Eastern Health, Monash University and an unrestricted investigator-initated research grant from Amgen Australia Pty. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.