Osteogenic and Anti-Osteoclastogenic Properties of the Curcuminoid, CRE-Bin: Ex Vivo Experiments

This research study was designed specifically to examine the influence of the curcuminoid (CRE-Bin) upon the osteogenic induction of MC3T3-E1 cells and upon the anti-osteoclastogenesis via RANKL-induced macrophage RAW 264.7 cells. Osteogenic induction and anti-osteoclastogenesis mechanisms for CRE-Bin-induced MC3T3-E1 cells and against RANKL-induced macrophage RAW 264.7 cells were investigated involving cell viability, ALP activity, Alizarin red S, Real-time PCR, TRAP content, cathepsin K expression, morphological changes, immunocytochemistry and signalling pathways analysis. CRE-Bin increased the viability of cells and induced ALP activity in MC3T3-E1 cells, hADSC, and also in C3H10T1/2, while calcium deposition in cells was also revealed. This resulted in a raised level of expression of mRNA in the Bmp-2, Runx2 and Collagen 1a osteoblast differentiation genes. Treatment with CRE-Bin upregulated the Wnt/β-catenin signalling circuit. Furthermore, CRE-Bin inhibited the formation of osteoclasts triggered by the receptor activator of the nuclear factor-κB ligand (RANKL) in RAW 264.7 cells. CRE-Bin increased F4/80 content, and consequently lowered TRAP content along with the osteoclast-specific gene, cathepsin K. RANKL-induced NF-κB and Akt components were inhibited by treatment with CRE-Ter, for example phospho-NF-κB-p65 and phospho-Akt proteins. From these results, one can infer that osteoporosis might in future be treated by novel drugs using CRE-Bin.