Novel 2,7-Diazaspiro[4,4]nonane Derivatives to Inhibit Mouse and Human Osteoclast Activities and Prevent Bone Loss in Ovariectomized Mice without Affecting Bone Formation

ABSTRACT Morroniside exerts a proosteogenic effect, which can prevent bone loss. However, the detailed mechanism underlying Morroniside-regulated bone formation is unclear. Morroniside can maintain cell homeostasis by promoting PI3K/Akt/mTOR signaling. The purpose of this study is to explore the significance of PI3K/Akt/mTOR signaling in Morroniside-regulated osteogenesis. The results showed that Morroniside promoted the activities of PI3K, Akt, and mTOR in osteoblast precursor MC3T3-E1. The differentiation of MC3T3-E1 to mature osteoblasts promoted by Morroniside can be reversed by the pharmacological inhibition of PI3K or mTOR. Importantly, in the presence of Morroniside, the osteoblast differentiation suppressed by PI3K inhibitor was reversed by mTOR overexpression. In vivo assays showed that in bone tissue of ovariectomized mice, Morroniside-enhanced osteoblast formation was reversed by the pharmacological inhibition of PI3K or mTOR. In conclusion, Morroniside can promote the osteogenesis through PI3K/Akt/mTOR signaling, which provides a novel clue for the strategy of Morroniside in treating osteoporosis.

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3579 The role of CypD/mPTP during osteogenic differentiation - a potential target to prevent bone loss

Journal of Clinical and Translational Science ◽

10.1017/cts.2019.58 ◽

2019 ◽

Vol 3 (s1) ◽

pp. 24-24

Author(s):

Rubens Sautchuk ◽

Brianna H. Shares ◽

Roman A. Eliseev

Keyword(s):

Bone Loss ◽

Bone Formation ◽

Mitochondrial Function ◽

Ectopic Bone Formation ◽

Regulatory Mechanisms ◽

Mrna Levels ◽

Prevent Bone Loss ◽

Ectopic Bone

OBJECTIVES/SPECIFIC AIMS: The study aims to further investigate how cyclophilin D (CypD), the key mPTP opening regulator, affects BMSCs fate and to determine potential regulatory mechanisms involved in CypD regulation during osteogenesis. METHODS/STUDY POPULATION: We evaluated CypD mRNA expression in mouse BMSCs and in osteogenic-like (OL) cells during the course of OB differentiation. CypD protein level was also probed. Moreover, BMSCs had their mPTP activity recorded during osteoinduction. We further analyzed the effect of CypD genetic deletion on osteogenesis in vitro and in vivo. For our in vivo model, we performed the ectopic bone formation assay to asses differences in ossicle formation when CypD KO BMSCs were transplanted compared to wild type littermate BMSCs. In our in vitro model, we transfected OL cells with either CypD gain of function or CypD loss of function vector and measured their osteogenic differentiation potential. Additionally, we treated BMSCs with CypD inhibitor and compare to non-treated BMSCs for mineralization level. To determine potential regulatory mechanisms involved in CypD regulation, we analyzed the CypD gene (Ppif) promoter for potential transcription factor (TF) binding sites and found multiple Smad-binding elements within this promoter. Smads (Smad1, 5, 8) are TFs downstream from Bone Morphogenic Protein (BMP) signaling pathway that transmit cell differentiation signaling, and exert either activating or inhibitory effects on a variety of genes. We also transfect OL cells with Smad1 vector and analyzed for CypD mRNA levels. RESULTS/ANTICIPATED RESULTS: - Our data showed that CypD mRNA levels decreased in both primary cells and OL cells at day 7 and day 14 in osteogenic media. - Osteogenic induction also decreased mPTP activity. - In vivo ectopic bone formation assay showed increased ossicle fo DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that downregulation of CypD increases OB differentiation due to improved OxPhos activity led by mPTP closure. Our results corroborate reports of CypD downregulation and mPTP closure during neuronal differentiation in developing rat brains as well as in cardiomyocyte differentiation in developing mouse hearts. Our studies also suggest a yet unknown mechanism linking differentiation signaling with mitochondrial function – BMP/Smad mediated downregulation of CypD transcription. As initially mentioned, in a previous study, our lab showed that CypD KO mice present higher mitochondrial function and osteogenicity in aged BMSCs and less osteoporosis burden. Taken together, these results suggest that CypD can be a potential target to prevent bone loss in aging.

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Magnesium Picolinate Improves Bone Formation by Regulation of RANK/RANKL/OPG and BMP-2/Runx2 Signaling Pathways in High-Fat Fed Rats

Nutrients ◽

10.3390/nu13103353 ◽

2021 ◽

Vol 13 (10) ◽

pp. 3353

Author(s):

Emre Sahin ◽

Cemal Orhan ◽

Tansel Ansal Balci ◽

Fusun Erten ◽

Kazim Sahin

Keyword(s):

Bone Loss ◽

Bone Formation ◽

High Fat Diet ◽

Male Rats ◽

High Fat ◽

Mineral Density ◽

Protein Levels ◽

Prevent Bone Loss ◽

Osteogenic Protein ◽

Affect Bone Metabolism

Magnesium (Mg) deficiency may affect bone metabolism by increasing osteoclasts, decreasing osteoblasts, promoting inflammation/oxidative stress, and result in subsequent bone loss. The objective of the present study was to identify the molecular mechanism underlying the bone protective effect of different forms of Mg (inorganic magnesium oxide (MgO) versus organic magnesium picolinate (MgPic) compound) in rats fed with a high-fat diet (HFD). Forty-two Wistar albino male rats were divided into six group (n = 7): (i) control, (ii) MgO, (iii) MgPic, (iv) HFD, (v) HFD + MgO, and (vi) HFD + MgPic. Bone mineral density (BMD) increased in the Mg supplemented groups, especially MgPic, as compared with the HFD group (p < 0.001). As compared with the HFD + MgO group, the HFD + MgPic group had higher bone P (p < 0.05) and Mg levels (p < 0.001). In addition, as compared to MgO, MgPic improved bone formation by increasing the levels of osteogenetic proteins (COL1A1 (p < 0.001), BMP2 (p < 0.001), Runx2 (p < 0.001), OPG (p < 0.05), and OCN (p < 0.001), IGF-1 (p < 0.001)), while prevented bone resorption by reducing the levels of RANK and RANKL (p < 0.001). In conclusion, the present data showed that the MgPic could increase osteogenic protein levels in bone more effectively than MgO, prevent bone loss, and contribute to bone formation in HFD rats.

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Effects of Estrogen Receptor and Wnt Signaling Activation on Mechanically Induced Bone Formation in a Mouse Model of Postmenopausal Bone Loss

International Journal of Molecular Sciences ◽

10.3390/ijms21218301 ◽

2020 ◽

Vol 21 (21) ◽

pp. 8301

Author(s):

Astrid Liedert ◽

Claudia Nemitz ◽

Melanie Haffner-Luntzer ◽

Fabian Schick ◽

Franz Jakob ◽

...

Keyword(s):

Estrogen Receptor ◽

Bone Loss ◽

Bone Formation ◽

Bone Mass ◽

Postmenopausal Bone Loss ◽

Paracrine Factors ◽

Treatment Of Osteoporosis ◽

Ovariectomized Mice ◽

Underlying Mechanisms ◽

Signaling Activation

In the adult skeleton, bone remodeling is required to replace damaged bone and functionally adapt bone mass and structure according to the mechanical requirements. It is regulated by multiple endocrine and paracrine factors, including hormones and growth factors, which interact in a coordinated manner. Because the response of bone to mechanical signals is dependent on functional estrogen receptor (ER) and Wnt/β-catenin signaling and is impaired in postmenopausal osteoporosis by estrogen deficiency, it is of paramount importance to elucidate the underlying mechanisms as a basis for the development of new strategies in the treatment of osteoporosis. The present study aimed to investigate the effectiveness of the activation of the ligand-dependent ER and the Wnt/β-catenin signal transduction pathways on mechanically induced bone formation using ovariectomized mice as a model of postmenopausal bone loss. We demonstrated that both pathways interact in the regulation of bone mass adaption in response to mechanical loading and that the activation of Wnt/β-catenin signaling considerably increased mechanically induced bone formation, whereas the effects of estrogen treatment strictly depended on the estrogen status in the mice.

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Glucocorticoid-induced osteoporosis

Arquivos Brasileiros de Endocrinologia & Metabologia ◽

10.1590/s0004-27302006000400024 ◽

2006 ◽

Vol 50 (4) ◽

pp. 793-801 ◽

Cited By ~ 13

Author(s):

Luiz Henrique de Gregório ◽

Paulo G. Sampaio Lacativa ◽

Ana Cláudia C. Melazzi ◽

Luis Augusto Tavares Russo

Keyword(s):

Vitamin D ◽

Bone Loss ◽

Bone Formation ◽

Preventive Measures ◽

Prevent Bone Loss ◽

Antiresorptive Agents ◽

Increased Risk ◽

Prevention And Treatment ◽

Small Doses ◽

Risk Patients

Glucocorticoid-induced osteoporosis is the most frequent cause of secondary osteoporosis. Glucocorticoids cause a rapid bone loss in the first few months of use, but the most important effect of the drug is suppression of bone formation. The administration of oral glucocorticoid is associated with an increased risk of fractures at the spine and hip. The risk is related to the dose, but even small doses can increase the risk. Patients on glucocorticoid therapy lose more trabecular than cortical bone and the fractures are more frequent at the spine than at the hip. Calcium, vitamin D and activated forms of vitamin D can prevent bone loss and antiresorptive agents are effective for prevention and treatment of bone loss and to decrease fracture risk. Despite the known effects of glucocorticoids on bone, only a few patients are advised to take preventive measures and treat glucocorticoid-induced osteoporosis.

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The natural polyamines spermidine and spermine prevent bone loss through preferential disruption of osteoclastic activation in ovariectomized mice

British Journal of Pharmacology ◽

10.1111/j.1476-5381.2012.01856.x ◽

2012 ◽

Vol 166 (3) ◽

pp. 1084-1096 ◽

Cited By ~ 30

Author(s):

Tomomi Yamamoto ◽

Eiichi Hinoi ◽

Hiroyuki Fujita ◽

Takashi Iezaki ◽

Yoshifumi Takahata ◽

...

Keyword(s):

Bone Loss ◽

Prevent Bone Loss ◽

Ovariectomized Mice

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Kanamycin inhibits daidzein metabolism and abilities of the metabolites to prevent bone loss in ovariectomized mice

BMC Research Notes ◽

10.1186/s13104-016-2139-7 ◽

2016 ◽

Vol 9 (1) ◽

Author(s):

Shin-ichi Katsumata ◽

Maiko Fujioka ◽

Shungo Fujii ◽

Ken Takeda ◽

Yoshiko Ishimi ◽

...

Keyword(s):

Bone Loss ◽

Prevent Bone Loss ◽

Ovariectomized Mice

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Monotropein isolated from the roots of Morinda officinalis increases osteoblastic bone formation and prevents bone loss in ovariectomized mice

Fitoterapia ◽

10.1016/j.fitote.2016.03.013 ◽

2016 ◽

Vol 110 ◽

pp. 166-172 ◽

Cited By ~ 24

Author(s):

Zhiguo Zhang ◽

Qiaoyan Zhang ◽

Hua Yang ◽

Wei Liu ◽

Naidan Zhang ◽

...

Keyword(s):

Bone Loss ◽

Bone Formation ◽

Ovariectomized Mice ◽

Osteoblastic Bone Formation

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Human Placenta-Derived Adherent Cells Prevent Bone loss, Stimulate Bone formation, and Suppress Growth of Multiple Myeloma in Bone

Stem Cells ◽

10.1002/stem.572 ◽

2011 ◽

Vol 29 (2) ◽

pp. 263-273 ◽

Cited By ~ 57

Author(s):

Xin Li ◽

Wen Ling ◽

Angela Pennisi ◽

Yuping Wang ◽

Sharmin Khan ◽

...

Keyword(s):

Multiple Myeloma ◽

Bone Loss ◽

Bone Formation ◽

Human Placenta ◽

Adherent Cells ◽

Stimulate Bone Formation ◽

Prevent Bone Loss

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A quinoxaline-based compound ameliorates bone loss in ovariectomized mice

Experimental Biology and Medicine ◽

10.1177/15353702211032133 ◽

2021 ◽

pp. 153537022110321

Author(s):

Ying Zhou ◽

Xiaoyan Xue ◽

Yanyan Guo ◽

Huan Liu ◽

Zheng Hou ◽

...

Keyword(s):

Weight Gain ◽

Bone Loss ◽

Bone Formation ◽

Marker Gene ◽

Mice Model ◽

Ovariectomized Mice ◽

Bone Formation Markers ◽

Small Molecule Compound ◽

Glucagon Like Peptide 1 ◽

Turnover Markers

DMB (6,7-dichloro-2-methylsulfonyl-3-Ntert-butylaminoquinoxaline) is a quinoxaline-based compound that has been investigated as a glucagon-like peptide-1 receptor (GLP-1R) agonist. To clarify anti-osteoporosis effect of DMB, an osteoporotic mice model was established by ovariectomy (OVX) operation. The OVX mice were given intraperitoneally DMB, exendin-4 (EX-4), or 17β-estradiol (E2) for two months. Then bone mass and structure, and bone morphometric parameters were examined by micro-CT. Weight gain and food consumption, bone turnover markers, and biomechanical strength of the femur were tested, and bone histomorphometry was analyzed. The food intake and weight gain was obviously reduced by E2 or EX-4, but not DMB. However, DMB or EX-4 treatment obviously inhibited skeletal deterioration and enhanced bone strength. The improvement involved in the increased osteoblast number and level of bone formation markers, and reduced osteoclasts number and level of bone resorption markers. In addition, DMB was found to stimulate osteoblastogenesis-related marker gene expression. These results demonstrated that DMB ameliorated bone loss mainly via induction of bone formation, which suggests that the small molecule compound might be applied to the management of postmenopausal osteoporosis.

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