Role of Lipopolysaccharide and RANKL in Osteoclastogenesis: Potential Inhibitory Effects of C-Phycocyanin on the Respective Molecular Pathways of Osteoclastogenesis
AuthorAl Qranei, Mohammed
AdvisorChellaiah, Meenakshi A.
MetadataShow full item record
AbstractMany skeletal diseases are characterized by excessive bone loss. Bone loss is mediated by osteoclasts, which are differentiated from cells of the monocyte/macrophage lineage upon stimulation of two indispensable factors, the RANKL and M-CSF. Lipopolysaccharide (LPS), a bacterial pathogenic factor, has also been shown to engage in osteoclastogenesis during inflammatory events actively. C-phycocyanin (C-PC) is a phycobiliprotein found in the blue-green algae that showed many therapeutic effects, including anti-arthritic and anti-inflammatory properties. However, the exact mechanism by which LPS regulates osteoclastogenesis and also the impact of C-PC on osteoclastogenesis needs further elucidations. We studied the osteoclast differentiation process in vitro using RAW 264.7 macrophage cell line. First, we showed that LPS induced osteoclastogenesis in RANKL-primed cells in vitro. LPS elicited osteoclastogenic mechanism by signaling through the TLR4 receptor, which is expressed in osteoclast precursors. Here we also found that TNF-α secreted by osteoclast precursors in response to TLR4 stimulation regulated the processes of osteoclastogenesis via TNF-R signaling. Second, we tested the inhibitory effect of C-PC on osteoclastogenesis. We showed here that C-PC strongly inhibited the early stage of osteoclast differentiation, thus significantly suppressing RANKL- and LPS- mediated osteoclastogenesis. Nonetheless, osteoblast differentiation and activity were not affected by C-PC. Reactive oxygen species (ROS) are generated during RANKL-mediated osteoclast differentiation. While studying the possible mechanisms of osteoclast differentiation, we found that C-PC a) attenuated RANKL-induced ROS; and b) interfered with RANKL-stimulated NF-κB signaling by preventing the degradation of cytosolic IκB-α; subsequently, these resulted in the loss of sequential activation of the osteoclastogenic downstream markers such as c-Fos and NFATc1. We propose that a unique mechanism of osteoclastogenesis is mediated by bacterial LPS that can be targeted during inflammatory-mediated bone loss. Also, C-PC could potentially be used as a therapeutic compound in osteolytic diseases caused by osteoclast activation without affecting osteoblast function.
Biomedical Sciences-Dental School
University of Maryland, Baltimore