Inositol Hexakisphosphate Inhibits Osteoclastogenesis on RAW 264.7 Cells and Human Primary Osteoclasts
Figure 6
Effect of inositol hexakisphosphate (IP6) on human peripheral blood mononuclear cells (PBMNC) osteoclastogenesis and resorption activity on mature osteoclasts cells derived from human PBMNC.
(A) Effect of IP6 treatment on the generation of multinucleated TRAP-positive cells (OCL). Human PBMNC, from three different donors, were cultured for 21 days with RANKL# (left) and with RANKL# plus 1 µM IP6 for 21 days (right). RANKL# refers to cells treated with RANKL and also with M-CSF and dexamethasone as described in the Materials & Methods section. Representative images are shown. (B) Number of multinucleated TRAP-positive cells with 3 or more nuclei (OCL) generated from human PBMNC treated with IP6. (C) Bone resorption ability of human PBMNC treated with RANKL# alone (left) and with 1 µM of IP6 during osteoclastogenesis (right) was evaluated by resorption pit assay on dentine discs (D) Percentage of resorbed area by human PBMNC treated with or without IP6 (1 µM) for each donor. (E) Representative images of bone resorption by mature osteoclasts treated with RANKL# alone (left) and with IP6 (right). (F) Percentage of resorbed dentin area by mature osteoclasts. Three different donors were used and the experiments were performed in triplicate, different shaded bars represent different donors, for each donor the data were presented as mean values ± SEM. Significant differences were assessed by paired t- test: *p≤0.05 versus RANKL# treated cells.