In vertebrates, melatonin is primarily secreted from the pineal gland but it affects various biological processes including the sleep-wake cycle, vasomotor control, immune system and bone homeostasis. Melatonin has been known to promote osteoblast differentiation and bone maturation, but a direct role of melatonin on osteoclast differentiation is still elusive. The present study investigated the effect of melatonin on the differentiation of macrophages to osteoclasts. The presence of melatonin significantly reduced receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis and the siRNA-mediated knockdown of the melatonin receptor failed to overcome the anti-osteoclastogenic effect of melatonin. Although melatonin treatment did not affect the phosphorylation of extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK), it markedly inhibited the activation of NF-κB and subsequent induction of nuclear factor of activated T cell cytoplasmic 1(NFATc1). Thus, our results suggest that melatonin could suppress osteoclast differentiation through downregulation of NF-κB pathway with concomitant decrease in the NFATc1 transcription factor induction. Furthermore, melatonin seems to have an anti-osteoclastogenic effect independent of plasma membrane melatonin receptors. In addition to previously reported properties of melatonin, our study proposes another aspect of melatonin and bone homeostasis.

Bisphosphonates are one of the most widely used synthetic pyrophosphate analogues for the treatment of bone resorbing diseases such as osteoporosis, multiple myeloma, and bone metastases. Although the therapeutic usefulness of bisphosphonates mainly depends on their anti-osteoclastogenic effect, a severe side-effect of bisphosphonates called bisphosphonate-related osteonecrosis of the jaw (BRONJ) could not be explained by the anti-osteoclastogenic effect of bisphosphonates. In the present study, we have evaluated the changes in osteoclastogenesis- or osteoblastogenesis-supporting activities of osteocytes induced by bisphosphonates. Zoledronate, a nitrogen-containing bisphosphonate, markedly increased both the receptor activator of nuclear factor kB ligand (RANKL) as well as sclerostin in osteocyte-like MLO-Y4 cells, which were functionally revalidated by osteoclast/osteoblast generating activities of the conditioned medium obtained from zoledronate-treated MLO-Y4 cells. Of note, the zoledronate treatment-induced upregulation of the RANKL expression was mediated by autocrine interleukin-6 (IL-6) and subsequent activation of the signal transducer and activator of transcription 3 (STAT3) pathway. These results were evidenced by the blunted RANKL expression in the presence of a Janus activated kinase (JAK2)/STAT3 inhibitor, AG490. Also, the osteoclastogenesis-supporting activity was significantly decreased in zoledronate-treated MLO-Y4 cells in the presence of IL-6 neutralizing IgG compared to that of the control IgG. Thus, our results show previously unanticipated effects of anti-bone resorptive bisphosphonate and suggest a potential clinical importance of osteocytes in BRONJ development.

The present study aimed to compare the bone-regeneration capacity of porcine-derived xenografts to bovine-derived xenografts in the rat calvarial defect model. The observation of surface morphology and in vitro cell studies were conducted prior to the animal study. Defects with a diameter of 8 mm were created in calvaria of 20 rats. The rats were randomly treated with porcine-derived (Bone-XP group) or bovine-derived xenografts (Bio-Oss group) and sacrificed at 4 and 8 weeks after surgery. The new bone regeneration was evaluated by micro-computed tomography (μCT) and histomorphometric analyses. In the cell study, the extracts of Bone-XP and Bio-Oss showed a positive effect on the regulation of osteogenic differentiation of human mesenchymal stem cells (hMSCs) without cytotoxicity. The new bone volume of Bone-XP (17.52 ± 3.78% at 4 weeks and 32.09 ± 3.51% at 8 weeks) was similar to that of Bio-Oss (11.6 ± 3.88% at 4 weeks and 25.89 ± 7.43% at 8 weeks) (p > 0.05). In the results of new bone area, there was no significant difference between Bone-XP (9.08 ± 5.47% at 4 weeks and 25.22 ± 13.56% at 8 weeks) and Bio-Oss groups (5.83 ± 2.56% at 4 weeks and 21.68 ± 11.11% at 8 weeks) (p > 0.05). It can be concluded that the porcine-derived bone substitute may offer a favorable cell response and bone regeneration similar to those of commercial bovine bone mineral.