Inhibitory effects of osteoprotegerin on osteoclast formation and function under serum-free conditions

J Vet Sci. 2013;14(4):405-12. doi: 10.4142/jvs.2013.14.4.405. Epub 2013 Jun 30.

Abstract

The purpose of this study was to determine whether osteoprotegerin (OPG) could affect osteoclat differentiation and activation under serum-free conditions. Both duck embryo bone marrow cells and RAW264.7 cells were incubated with macrophage colony stimulatory factor (M-CSF) and receptor activator for nuclear factor kB ligand (RANKL) in serum-free medium to promote osteoclastogenesis. During cultivation, 0, 10, 20, 50, and 100 ng/mL OPG were added to various groups of cells. Osteoclast differentiation and activation were monitored via tartrate-resistant acid phosphatase (TRAP) staining, filamentous-actin rings analysis, and a bone resorption assay. Furthermore, the expression osteoclast-related genes, such as TRAP and receptor activator for nuclear factor κB (RANK), that was influenced by OPG in RAW264.7 cells was examined using real-time polymerase chain reaction. In summary, findings from the present study suggested that M-CSF with RANKL can promote osteoclast differentiation and activation, and enhance the expression of TRAP and RANK mRNA in osteoclasts. In contrast, OPG inhibited these activities under serum-free conditions.

Keywords: activation; differentiation; osteoclast; osteoprotegerin; serum-free.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Phosphatase / genetics
  • Acid Phosphatase / metabolism
  • Animals
  • Avian Proteins / pharmacology*
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism*
  • Cells, Cultured
  • Ducks
  • Embryo, Nonmammalian / drug effects
  • Embryo, Nonmammalian / metabolism
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Macrophage Colony-Stimulating Factor / metabolism
  • Osteoclasts / cytology
  • Osteoclasts / drug effects*
  • Osteoclasts / metabolism*
  • Osteoprotegerin / pharmacology*
  • RANK Ligand / metabolism
  • Real-Time Polymerase Chain Reaction
  • Receptor Activator of Nuclear Factor-kappa B / genetics
  • Receptor Activator of Nuclear Factor-kappa B / metabolism
  • Tartrate-Resistant Acid Phosphatase

Substances

  • Avian Proteins
  • Isoenzymes
  • Osteoprotegerin
  • RANK Ligand
  • Receptor Activator of Nuclear Factor-kappa B
  • Macrophage Colony-Stimulating Factor
  • Acid Phosphatase
  • Tartrate-Resistant Acid Phosphatase