Background : Osteoclasts are differentiated from the monocytes/macrophages of hematopoietic cells, that excessive activities of bone-resorbing giant cells leads to pathological bone diseases such as osteoporosis (contained rheumatoid arthritis and autoimmune arthritis). Therefore, it is very important to suppress loss of bone mass by deactivation of osteoclast differentiation. In this context, we evaluated for the effects of black ginseng (BG) extract on TRAP activity, proliferation and differentiation in RANKL-induced osteoclastic RAW264.7 cells. Methods and Results : The aim of this study is to figure out the potential anti-osteoporosis effects and the underlying mechanism of BG extract in RANKL-induced osteoclastic RAW264.7 cells. The ginsenoside Rg3, Rg5, Rk1 and Rh4 content of BG was increased more than Red ginseng (RG). The extracts of BG markedly reduced the activity of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from osteoclastic RAW264.7 cells, without cytotoxicity. BG clearly inhibited RANKL-induced osteoclast differentiation by decreased calcitonin and TRAP (p < 0.01). Furthermore, ginseonside Rg5 and Rk1 significantly inhibited TRAP activity in formation of osteoclastic differentiation (p < 0.01). It is also found that Ginseonside Rg5 and Rk1 mixture more inhibits osteoclast differentiation activity. Conclusion : Our results suggest that Black ginseng extract has an anti-osteoporosis effects in bone disease when administered as a food supplement and has potential as a therapeutic agent for osteoporosis.

This study investigated the effects of pressurized steam-treated Corni Frutus (PSC) extract on osteoblast differentiation and osteoclast formation. The osteoblast differentiation effect of the extract was evaluated by measuring cellular alkaline phosphatase (ALP) activity, cell matrix ALP staining, alizarin Red S staining and von Kossa staining on proliferating MC3T3-E1 osteoblast cells. The results confirmed that ALP activity, cell matrix ALP staining, alizarin Red S staining and von Kossa staining were all increased as proliferation increased from 1 to 14 days, without cytotoxicity. The osteoclast formation effect of the PSC extract was evaluated by measuring the cellular tartrate-resistant acid phosphatase (TRAP) activity and cell matrix TRAP staining on receptor activator of nuclear factor kappa-B ligand (RANKL)-induced RAW264.7 osteoclast cells. Treating RAW264.7 cells with RANKL for 7 days increased matrix staining for TRAP and cellular TRAP activity. The PSC extract decreased these changes in a concentration-dependent manner. Therefore, PSC is expected to be a natural source for developing health functional foods and medicinal agents to prevent bone-related diseases, such as osteoporosis, by increasing osteoblast differentiation and reducing osteoclast activity.

Background : Capsaicin is an active component of chili peppers, which are plants belonging to the genus Capsicum. Research reported capsaicin has antioxidant and anti-inflamatory activity and osteoclast lineages are very susceptible to oxidative stress, as osteoclasts are produced by increased-generation of intracellular ROS and osteoclasts are activated by ROS. Therefore, our study was evaluated the influence of intracellular oxidative stress such as increased ROS level on RANKL-mediated osteoclast differenciation. Methods and Results : Capsaicin showed a good free radical scavenging activity at in-vitro antioxidant activity. The inhibitory effect of osteoclast differentiation on capsaicin was confirmed. Osteoclast differentiation from murine macrophage RAW 264.7 cells was induced by RANKL. The effect of capsaicin on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclast differentiation was demonstrated using a tartrate-resistant acid phosphatase (TRAP) assay and TRAP staining. Capsaicin showed an inhibitory effect on TRAP activity. The TRAP staining showed that the number of TRAP positive osteoclasts was reduced in capsaicin-treated cells. Conclusion : Capsaicin revealed an inhibitory effect in osteoclast differentiation induced by RANKL. These results suggest that capsaicin may have a beneficial effect for the prevention or treatment of osteoclast caused bone diseases such as osteoporosis.

Background : Osteoclasts as multinucleated cells originate from hematopoietic monocyte/ macrophage precursor cell, shows the bone absorption through the commitment, differentiation, fusion, and bone resorption stages by regulation of M-CSF and RANKL. It has been reported a significant negative correlation between the increase of oxidative stress and the bone density, and when RANKL reaction to the osteoclasts precursor cells is mainly generated ROS is due to increased activity of NADPH oxidase1 (NOX1), and these ROS act as a factor which promotes osteoclasts differentiation. Thus, RANKL signaling process is important that excessive osteoclast formation and differentiation inhibited through the regulation of each step. Methods and Results : F3570 ethanol extract showed relatively high activity at in-vitro antioxidant activity. F3570 water extract inhibited ROS generation in RAW 264.7 cells stimulated with H2O2 and RANKL, even at low concentrations. The inhibitory effect of osteoclast differentiation on F3570 water extract was confirmed that shown through NF-κB pathway, MAPK pathway including ERK and JNK. F3570 ethanol extract is considered to be regulated by the p38 MAPK and the other signaling pathway. Also, F3570 both water and ethanol extract were significantly reduced gene expression such as TRAP, calcitonin receptors and integrin β3 of RANKL-induced mature osteoclast in the bone resorption stage. Conclusion : Through this study, F3570 extract revealed an outstanding inhibitory effect and signaling mechanisms in osteoclast differentiation induced by RANKL. These results suggest that F3570 is bone diseases associated with aging or osteoporosis caused by menopause in an aging society is expected to be a superior candidate for the treatment or the prevention

Osteoporosis induces a bone mineral density loss due to imbalance of bone homeostasis that is achieved by osteoclasts (which are involved in bone resorption) and osteoblasts (which are involved in bone formation). Thus, this study was performed to evaluate the effects of hot water extract of the Achyranthes bidentata Blume (ABB) and Panax ginseng (Gin) on osteoclast and osteoblast differentiation. In this study, there was no cytotoxicity by ABB, 50 and 100 μg/ml of Gin significantly decreased cell viability of RANKL-induced osteoclast in RAW264.7 cell (p < 0.01). But, it was 50 μg/ml of ABB and Gin mixtures increased due to protective action of ABB. Furthermore, Gin contained groups (Gin, ABB and Gin mixtures) were inhibitory effects on osteoclast differentiation and bone resorption, and increased in osteoblast differentiation activity. Gin clearly inhibited RANKL-induced osteoclast differentiation by decreased calcitonin and TRAP (p < 0.01). Also, these extracts significantly increased calcium accumulation formation of osteoblastic differentiation reagents-induced osteoblast in MC3T3-E1 cell (p < 0.05). These results suggest that ABB and Gin mixtures may be a potential as drug for the treatment of osteoporosis.