Effect of BioAggregate on Receptor Activator of Nuclear Factor-Kappa B Ligand-induced Osteoclastogenesis from Murine Macrophage Cell Line In Vitro

J Endod. 2015 Aug;41(8):1265-71. doi: 10.1016/j.joen.2015.03.021. Epub 2015 May 12.

Abstract

Introduction: This study aimed to investigate the effect of BioAggregate, a calcium silicate-based nanoparticulate bioceramic, on the regulation of receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclast differentiation and bone resorption in vitro, as well as to delineate the underlying molecular mechanism. The performance of BioAggregate was compared with that of ProRoot mineral trioxide aggregate (MTA).

Methods: Cells of a murine macrophage cell line RAW 264.7 were treated with various concentrations of BioAggregate and MTA extracts. Cytotoxicity of material extracts was evaluated with Cell Counting Kit-8 assay. RANKL-induced osteoclast differentiation and function were assessed with tartrate-resistant acid phosphatase staining, F-actin staining, and lacunar resorption pits assay. The mRNA expression associated with osteoclast function was detected with quantitative real-time polymerase chain reaction. Related molecular signaling pathways were investigated with Western blot and immunofluorescence.

Results: BioAggregate extracts dose-dependently inhibited RANKL-induced osteoclast formation and resorption capacity without evident cytotoxicity. RAW 264.7 cells exposed to BioAggregate extracts also presented a decrease in RANKL-stimulated mRNA expression of osteoclast-related genes and transcription factors. Moreover, cells treated with BioAggregate extracts exhibited attenuated TRAF6 expression, suppressed mitogen-activated protein kinase signaling, and decreased nuclear translocation of NFATc1 and c-Fos in the presence of RANKL. Comparable effects were induced by MTA.

Conclusions: BioAggregate and MTA exhibit comparable inhibitory effect on osteoclast differentiation and function in vitro, and our findings provide valuable insights into the mechanism of bioceramic-mediated anti-osteoclastogenic activity.

Keywords: BioAggregate; NFATc1; osteoclastogenesis; receptor activator for nuclear factor kappaB; root repair.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus / drug effects
  • Active Transport, Cell Nucleus / physiology
  • Animals
  • Bone Resorption / drug therapy
  • Bone Resorption / physiopathology
  • Calcium Hydroxide / pharmacology*
  • Calcium Hydroxide / toxicity
  • Cell Line
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Dose-Response Relationship, Drug
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Growth Substances / pharmacology*
  • Growth Substances / toxicity
  • Hydroxyapatites / pharmacology*
  • Hydroxyapatites / toxicity
  • Macrophages / drug effects*
  • Macrophages / physiology
  • Mice
  • NF-kappa B / metabolism*
  • NFATC Transcription Factors / metabolism
  • Osteogenesis / drug effects*
  • Osteogenesis / physiology
  • Proto-Oncogene Proteins c-fos / metabolism
  • RNA, Messenger / metabolism
  • Root Canal Filling Materials / pharmacology*
  • Root Canal Filling Materials / toxicity
  • Silicates / pharmacology*
  • Silicates / toxicity
  • TNF Receptor-Associated Factor 6 / metabolism

Substances

  • BioAggregate
  • Growth Substances
  • Hydroxyapatites
  • NF-kappa B
  • NFATC Transcription Factors
  • Nfatc1 protein, mouse
  • Proto-Oncogene Proteins c-fos
  • RNA, Messenger
  • Root Canal Filling Materials
  • Silicates
  • TNF Receptor-Associated Factor 6
  • Extracellular Signal-Regulated MAP Kinases
  • Calcium Hydroxide