title: Effect of BioAggregate on Receptor Activator of Nuclear Factor-Kappa B Ligand-induced Osteoclastogenesis from Murine Macrophage Cell Line In Vitro.
abstract: BACKGROUND
  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.
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
authors.forename: Lingxin
authors.initials: L
authors.lastname: Zhu
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Yan
authors.affiliation: State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China. Electronic address: phs301@vip.163.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Peng
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000502:physiology; D000818:Animals; D001862:Bone Resorption / Q000188:drug therapy / Q000503:physiopathology; D002126:Calcium Hydroxide / Q000494:pharmacology* / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology* / Q000633:toxicity; D006882:Hydroxyapatites / Q000494:pharmacology* / Q000633:toxicity; D008264:Macrophages / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D050778:NFATC Transcription Factors / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects* / Q000502:physiology; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012387:Root Canal Filling Materials / Q000494:pharmacology* / Q000633:toxicity; D017640:Silicates / Q000494:pharmacology* / Q000633:toxicity; D048029:TNF Receptor-Associated Factor 6 / Q000378:metabolism
chemical_list: C557009:BioAggregate; D006133:Growth Substances; D006882:Hydroxyapatites; D016328:NF-kappa B; D050778:NFATC Transcription Factors; C494005:Nfatc1 protein, mouse; D016760:Proto-Oncogene Proteins c-fos; D012333:RNA, Messenger; D012387:Root Canal Filling Materials; D017640:Silicates; D048029:TNF Receptor-Associated Factor 6; D048049:Extracellular Signal-Regulated MAP Kinases; D002126:Calcium Hydroxide
keywords: BioAggregate; NFATc1; osteoclastogenesis; receptor activator for nuclear factor kappaB; root repair