title: Effect of monoterpenes on the formation and activation of osteoclasts in vitro.
abstract: Monoterpenes, present in aromatic plants, are known to inhibit bone resorption in vivo. In this in vitro study, they inhibited the activation of osteoclasts only at high concentrations but inhibited the formation at much lower concentrations. Therefore, monoterpenes may act in vivo directly on osteoclastogenesis.
  
  
  BACKGROUND
  Monoterpenes are the major components of essential oils, which are formed in many plants. Typically, they are found in herbs and certain fruits. When fed to rats, they inhibit bone resorption by an unknown mechanism. In this study, their effect on the activity and formation of osteoclasts in vitro was studied.
  
  
  METHODS
  The effect of monoterpenes on the development of osteoclasts was studied in co-cultures of bone marrow cells and osteoblasts and in cultures of spleen cells grown with colony stimulating factor (CSF)-1 and RANKL. In cultures of primary osteoblasts, alkaline phosphatase activity and levels of mRNA encoding RANKL and osteoprotegerin (OPG) mRNA (RT-PCR), and in osteoblast and spleen cell cultures, lactate dehydrogenase activity, a measure of toxicity, were determined. The activity of isolated rat osteoclasts was determined by counting the osteoclasts with actin rings using histofluorometry.
  
  
  RESULTS
  The monoterpenes inhibited the formation of osteoclasts more strongly in co-cultures (> or = 1 microM) than in cultures of spleen cells (> or = 10 microM). They had a minor effect on osteoblasts. Toxic effects were not observed. The inhibition of the formation of osteoclasts was not reversed by the addition of farnesol and geranylgeraniol, excluding an effect of the monoterpenes through the mevalonate pathway. A high concentration of 1 mM was required to inhibit the activation of osteoclasts. This effect, shown for menthol and borneol, was reversible.
  
  
  CONCLUSIONS
  The results suggest that the monoterpenes inhibit bone resorption in vivo through a direct effect on the formation of osteoclasts acting mainly on the hemopoietic cells.
authors.affiliation: Bone Biology Group, Department of Clinical Research, University of Berne, Bern, Switzerland.
authors.forename: Silvia
authors.initials: S
authors.lastname: Dolder
authors.forename: Willy
authors.initials: W
authors.lastname: Hofstetter
authors.forename: Antoinette
authors.initials: A
authors.lastname: Wetterwald
authors.forename: Roman C
authors.initials: RC
authors.lastname: Mühlbauer
authors.forename: Rolf
authors.initials: R
authors.lastname: Felix
mesh_terms: D000818:Animals; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D039821:Monoterpenes / Q000494:pharmacology*; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects*; D010012:Osteogenesis / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D013154:Spleen / Q000166:cytology / Q000187:drug effects
chemical_list: D039821:Monoterpenes; D007770:L-Lactate Dehydrogenase