a-rankl-based-osteoclast-culture-assay-mouse-bone-marrow-to

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

Osteoclasts are unique bone-resorbing cells that differentiate from the monocyte/macrophage lineage of bone marrow. Dysfunction of osteoclasts may result ...

Shanghai Jiaotong University School of Medicine

A novel mesoporous magnesium-based cement (MBC) was fabricated by using the mixed powders of magnesium oxide, sodium dihydrogen phosphate, and mesoporous magnesium silicate (m-MS). The results indicate that the setting time and water absorption of the MBC increased as a function of increasing m-MS content, while compressive strength decreased. In addition, the degradability of the MBC in a solution of Tris-HCl and the ability of apatite formation on the MBC were significantly improved with the increase in m-MS content. In cell culture experiments, the results show that the attachment, proliferation, and alkaline phosphatase activity of the MC3T3-E1 cells on the MBC were significantly enhanced with the increase of the content of m-MS. It can be suggested that the MBC with good cytocompatibility could promote the proliferation and differentiation of the MC3T3-E1 cells. In short, our findings indicate that the MBC containing m-MS had promising potential as a new biocement for bone regeneration and repair applications.

Improvement of bioactivity, degradability, and cytocompatibility of biocement by addition of mesoporous magnesium silicate into sodium-magnesium phosphate cement.

Mammalian target of rapamycin complex 1 (mTORC1) is involved in anabolic metabolism in both osteoblasts and chondrocytes, but the role of mTORC1 in osteoclast biology in vivo remains to be elucidated. In this study, we showed that deletion of regulatory-associated protein of mTOR (Raptor) in osteoclasts led to an increase in bone mass with decreased bone resorption. Raptor-deficient bone marrow-derived macrophages exhibited lower mTORC1-S6K1 signaling and retarded osteoclast differentiation, as determined by the number of osteoclasts, tartrate-resistant acid phosphatase activity, and expression of osteoclast-specific genes. Enforced expression of constitutively active S6K1 rescued the impaired osteoclast differentiation in Raptor-deficient bone marrow-derived macrophages. Furthermore, pharmacological inhibition of mTORC1 signaling by rapamycin could also inhibit osteoclast differentiation and osteoclast-specific gene expression. Taken together, our findings demonstrate that mTORC1 plays a key role in the network of catabolic bone resorption in osteoclasts and may serve as a potential pharmacological target for the regulation of osteoclast activity in bone metabolic disorders.

Inactivation of Regulatory-associated Protein of mTOR (Raptor)/Mammalian Target of Rapamycin Complex 1 (mTORC1) Signaling in Osteoclasts Increases Bone Mass by Inhibiting Osteoclast Differentiation in Mice.

As the number of elderly orthodontic patients increases, the impact of postmenopausal osteoporosis on orthodontic tooth movement (OTM) has attracted a great deal of attention because OTM relies on alveolar bone remodeling. The question of whether OTM causes subsequent alveolar bone loss and is harmful to alveolar bone health under osteoporotic conditions remains to be answered. The present study aimed to clarify the influences of OTM on alveolar bone in osteoporotic rats. OTM was accelerated in ovariectomized (OVX) rats as a result of increased bone resorption in the pressure area. At the same time, anabolic bone formation was promoted in the tension area during OTM in OVX rats. Micro-CT analysis of alveolar bone revealed a decrease in BMD, BV/TV and Tb.Th. in the OTM group compared with that in non-OTM rats on day 21 of OTM, suggesting that OTM caused alveolar bone loss in OVX rats during OTM. However, the OTM-induced bone loss could be recovered 3 months after OTM in OVX rats. Thus, our findings suggest that increased osteogenesis may compensate for the increased bone resorption during and after OTM and enable effective accelerated OTM in OVX rats.

Force-induced increased osteogenesis enables accelerated orthodontic tooth movement in ovariectomized rats.

The mammalian target of rapamycin (mTOR)/regulatory-associated protein of mTOR (Raptor) pathway transmits and integrates different signals including growth factors, nutrients, and energy metabolism. Nearly all these signals have been found to play roles in skeletal biology. However, the contribution of mTOR/Raptor to osteoblast biology in vivo remains to be elucidated as the conclusions of recent studies are controversial. Here we report that mice with a deficiency of either mTOR or Raptor in preosteoblasts exhibited clavicular hypoplasia and delayed fontanelle fusion, similar to those found in human patients with cleidocranial dysplasia (CCD) haploinsufficient for the transcription factor runt-related transcription factor 2 (Runx2) or those identified in Runx2mice. Mechanistic analysis revealed that the mTOR-Raptor-S6K1 axis regulates Runx2 expression through phosphorylation of estrogen receptor α, which binds to Distal-less homeobox 5 (DLX5) and augments the activity of Runx2 enhancer. Moreover, heterozygous mutation of raptor in osteoblasts aggravates the bone defects observed in Runx2mice, indicating a genetic interaction between Raptor and Runx2. Collectively, these findings reveal that mTOR/Raptor signaling is essential for bone formation in vivo through the regulation of Runx2 expression. These results also suggest that a selective mTOR/Raptor antagonist, which has been developed for treatment of many diseases, may have the side effect of causing bone loss.

mTOR/Raptor signaling is critical for skeletogenesis in mice through the regulation of Runx2 expression.

                    
To investigate the relationship between clinical and histopathological characteristics and overall survival of patients with oral mucosal melanoma (OMM) without distal metastasis in order to provide predictive prognostic information of OMM.Ki67 expression was assessed by immunohistochemistry in 123 patients with OMM without distant metastases. The associations between Ki67 expression and clinical features and overall survival (OS) of patients were statistically analyzed. The Ki67 levels of the primary and recurrent lesions from 14 OMM patients were compared.Univariate analysis showed that tumor type and cervical lymph node (CLN) status, as well as Ki67 expression, were all correlated with survival. Cox proportional hazards regression analysis identified Ki67 expression and CLN status as independent prognostic factors in OMM patients. Further, we found that Ki67 expression was associated with clinical tumor type of OMM. Moreover, with a cut-off point of 20%, patients with lower Ki67 scores showed a survival advantage over those with higher Ki67 scores.Ki67 expression may be a useful pathological predictor of survival of OMM patients without distant metastases.

Ki67 Proliferation Index as a Histopathological Predictive and Prognostic Parameter of Oral Mucosal Melanoma in Patients without Distant Metastases.

The aim of the study was to study the specific morphological features of alveolar bone and compare it to femoral bone in rats.

Article	Year
Improvement of bioactivity, degradability, and cytocompatibility of biocement by addition of mesoporous magnesium silicate into sodium-magnesium phosphate cement. Journal of materials science. Materials in medicine\| PubMed ID: 26395363	2015
Inactivation of Regulatory-associated Protein of mTOR (Raptor)/Mammalian Target of Rapamycin Complex 1 (mTORC1) Signaling in Osteoclasts Increases Bone Mass by Inhibiting Osteoclast Differen... The Journal of biological chemistry\| PubMed ID: 27879318	2017
Force-induced increased osteogenesis enables accelerated orthodontic tooth movement in ovariectomized rats. Scientific reports\| PubMed ID: 28634415	2017
mTOR/Raptor signaling is critical for skeletogenesis in mice through the regulation of Runx2 expression. Cell death and differentiation\| PubMed ID: 28686577	2017
Ki67 Proliferation Index as a Histopathological Predictive and Prognostic Parameter of Oral Mucosal Melanoma in Patients without Distant Metastases. Journal of Cancer\| PubMed ID: 29151970	2017
The Specific Morphological Features of Alveolar Bone. The Journal of craniofacial surgery\| PubMed ID: 29498973	2018

Xuhui Ma

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