AaCDT  influences osteoclasts differentiation and cytokines response in BMC

Method: Recombinant AaCDT (rAaCDT) was added to BMC from C57/BL6 mice in α-MEM in the presence or absence of 50ng/ml RANKL. Culture medium was replaced every 48h. After 6 days, cell proliferation was estimated by MTT assay, cells were stained with TRAP and TRAP+ multinucleated cells were counted. IL-1β, IL-6, IL-10 and TNF-α concentration in supernatant were evaluated by ELISA. The data were analyzed by ANOVA followed by Tukey test (p < 0.05). 

Result: The addition of rAaCDT led to increased BMC viability. The number of multinucleated TRAP+ cells was significantly higher when BMC were treated with the highest concentration of rAaCDT (25 μg/ml) in the absence of RANKL. TNF-α production was not affected by the addition of rAaCDT. However IL-10 and IL-1-β production were increased with addition of high doses of rAaCDT, with or without RANKL. Interestingly, IL-6 production increased slightly but significantly, when rAaCDT was added in the absence of RANKL.  

Conclusion: CDT may influence osteoclasts differentiation by an alternative pathway, possibly due to the induction of cytokines such as IL-1β and IL-6, involved in osteoclastogenesis signaling.