Temporal differential effects of proinflammatory cytokines on osteoclastogenesis

• Published in: International journal of molecular medicine 2013-04, Vol.31 (4), p.769-777
• Main Authors: MOON, SU-JIN, AHN, INHYE E, JUNG, HYERIN, YI, HYOJU, KIM, JURYUN, KIM, YOUNGKYUN, KWOK, SEUNG-KI, PARK, KYUNG-SU, MIN, JUN-KI, PARK, SUNG-HWAN, KIM, HO-YOUN, JU, JI HYEON
• Format: Article
• Language: English
• Subjects: Apoptosis; Bone marrow; Cytokines; inflammation; interleukin-1β; interleukin-6; nuclear factor-κB; osteoclast; receptor activator nuclear factor-κB ligand; Tumor necrosis factor-TNF
• ISSN: 1107-3756; 1791-244X
• DOI: 10.3892/ijmm.2013.1269

Bone destruction and inflammation are closely linked. Cytokines play an important role in inflammatory bone destruction by upregulating the receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL). The direct role of cytokines that act in a non-RANKL-dependent manner has yet to be elucidated. The aim of this study was to investigate the direct osteoclastogenic properties of inflammatory cytokines at different time-points of osteoclastogenesis. Mouse bone marrow macrophages were stimulated with the macrophage colony-stimulating factor (M-CSF) and various concentrations of RANKL. Inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-17 and IL-23, were added to the culture system of osteoclastogenesis. Two time-points of cytokine treatment were set. The 'early' effect of each cytokine was investigated at the time of first RANKL treatment, whereas the 'late' effect was investigated 48 h after the first RANKL challenge. Osteoclast differentiation and function were assessed using an osteoclast marker [tartrate-resistant acid phosphatase (TRAP)] and by visualization of pit formation. A permissive level of RANKL was required for cytokine-associated osteoclastogenesis in all experiments. In the M-CSF/RANKL monocellular culture system, IL-1β enhanced and IL-6 decreased osteoclast formation in a dose-dependent manner, regardless of temporal differences. Other cytokines showed various responses according to the phase of osteoclast maturation and the concentration of each cytokine and RANKL. Furthermore, luciferase assays showed that both IL-1β and RANKL activated the NF-κB signaling pathway. Collectively, our data revealed that targeting IL-1β may be a promising strategy to inhibit inflammation-associated bone destruction and osteoporosis.