Leukotriene B₄ Loaded in Microspheres Inhibits Osteoclast Differentiation and Activation

Abstract

To investigate osteoclast formation in vivo and if leukotriene B₄ (LTB₄) loaded in microspheres (MS) could be used as a therapeutical strategy to promote a sustained delivery of the mediator and prevent osteoclast differentiation. Methods: In vivo, apical periodontitis was induced in mice to investigate osteoclast differentiation and signaling in absence of 5-lipoxygenase (5-LO). In vitro, LTB₄-MS were prepared using an oil-in-water emulsion solvent extraction-evaporation process. Characterization and efficiency of LTB₄ encapsulation were investigated. J774A.1 macrophages were cultured in the presence of monocyte colony-stimulating factor (M-CSF) and ligand for receptor activator of nuclear factor kappa B (RANKL) and then stimulated with LTB₄-MS. Cytotoxicity, in vitro MS-LTB₄ uptake, osteoclast formation and gene expression were measured. Results: We found that 5-LO negatively regulates osteoclastic formation in vivo during apical periodontitis development. In vitro, LTB₄-MS were up-taken by macrophages and were not cytotoxic to the cells. LTB₄-MS inhibited osteoclast formation and the synthesis of osteoclastogenic genes Acp5, Mmp9, Calcr and Ctsk. LTB₄-MS inhibited differentiation of macrophages into an osteoclastic phenotype and cell activation under M-CSF and RANKL stimulus.

Key Words:
Osteoclast; leukotriene B4; macrophages; differentiation; activation; microsphere