RNA isolation from alveolar bone and gene expression analysis of RANK, RANKL and OPG: A new tool to monitor bone remodeling and healing in different bone substitutes used for prosthetic rehabilitation

Canciani, Elena; Dellavia, Claudia; Marazzi, Monica Gioia; Augusti, Davide; Carmagnola, Daniela; Vianello, Elena; Canullo, Luigi; Galliera, Emanuela

doi:10.1016/j.archoralbio.2017.03.011

Cited by 11 publications

(10 citation statements)

References 14 publications

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“…Futhermore, differentiated osteoblasts also expressed RANKL and OPG, the markers of osteoblast-regulated osteoclast activation and bone resorption process [ 48 ]. RANKL is an essential regulator for activation, differentiation and survival of osteoclasts by binding to the specific RANK receptor, which is found on the surface of osteoclast precursors and mature osteoclasts [ 49 ]. On the other hand, OPG is a decoy receptor of RANKL and thus acts as an inhibitor of bone resorption.…”

Section: Discussionmentioning

confidence: 99%

Demethylbelamcandaquinone B (Dmcq B) Is the Active Compound of Marantodes pumilum var. alata (Blume) Kuntze with Osteoanabolic Activities

et al. 2018

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Phytoestrogens have attracted considerable attention for their potential in the prevention of postmenopausal osteoporosis. Recently, a phytoestrogen-rich herbal plant, Marantodes pumilum var. alata (Blume) Kuntze was reported to protect against bone loss in ovariectomized rat. However, the bioactive compound responsible for these effects and the underlying mechanism were not known. Through bioassay-guided isolation, demethylbelamcandaquinone B (Dmcq B) was isolated and identified from Marantodes pumilum var. alata leaf extract. In terms of its bone anabolic effects, Dmcq B was at par with 17β-estradiol (E2), in promoting the proliferation, differentiation and mineralization of osteoblast cells. Dmcq-B increased early differentiation markers, collagen content and enzymatic ALP activity. It was demonstrated to regulate BMP2 signaling pathway which further activated the transcription factor, osterix. Subsequently, Dmcq B was able to increase the osteocalcin expression which promoted matrix mineralization as evidenced by the increase in calcium deposition. Dmcq B also reduced the protein level of receptor activator of NF-κβ ligand (RANKL) and promoted osteoprotegerin (OPG) protein expression by osteoblast cells, therefore hastening bone formation rate by decreasing RANKL/OPG ratio. Moreover, Dmcq B was able to increase ER expression, postulating its phytoestrogen property. As the conclusion, Dmcq B is the active compound isolated from Marantodes pumilum var. alata leaves, regulating osteoanabolic activities potentially through the BMP2 and ER signaling pathways.

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Section: Discussionmentioning

confidence: 99%

Demethylbelamcandaquinone B (Dmcq B) Is the Active Compound of Marantodes pumilum var. alata (Blume) Kuntze with Osteoanabolic Activities

et al. 2018

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“…Total RNA was extracted using the TRI reagent method. RNA was extracted from the mycelium after growth in a liquid culture to ensure that a high yield was obtained with purity of RNA (Canciani et al, 2017). The A. flavus mycelia were treated with 0, 0.065, 0.13, and 0.26 mg/ml CA for 12 h, then collected, washed, and resuspended in sterile PBS.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Cinnamaldehyde, a Promising Natural Preservative Against Aspergillus flavus

Yang

Chen

et al. 2019

Front. Microbiol.

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The problem of food spoilage due to Aspergillus flavus (A. flavus) needs to be resolved. In this study, we found that the minimum inhibitory concentration of cinnamaldehyde (CA) that inhibited A. flavus was 0.065 mg/ml and that corn can be prevented from spoiling at a concentration of 0.13 mg/cm 3. In addition to inhibiting spore germination, mycelial growth, and biomass production, CA can also reduce ergosterol synthesis and can cause cytomembrane damage. Our intention was to elucidate the antifungal mechanism of CA. Flow cytometry, fluorescence microscopy, and western blot were used to reveal that different concentrations of CA can cause a series of apoptotic events in A. flavus, including elevated Ca 2+ and reactive oxygen species, decrease in mitochondrial membrane potential (ψ m), the release of cytochrome c, the activation of metacaspase, phosphatidylserine (PS) externalization, and DNA damage. Moreover, CA significantly increased the expression levels of apoptosis-related genes (Mst3, Stm1, AMID, Yca1, DAP3, and HtrA2). In summary, our results indicate that CA is a promising antifungal agent for use in food preservation.

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“…This trend of OPG is correlated with the decrease of RANK and RANKL in Sp7-Cre;SMS1 f/f ;SMS2 −/− mice. The RANKL/OPG ratio is considered a comprehensive indicator of prevailing activity of bone remodeling (Canciani et al 2017). The RANKL/OPG ratio is negative not only in Sp7-Cre;SMS1 f/f ;SMS2 −/− but in control mice as well.…”

Section: Discussionmentioning

confidence: 99%

Deficiency of sphingomyelin synthase 1 but not sphingomyelin synthase 2 reduces bone formation due to impaired osteoblast differentiation

Matsumoto

Hashizume

Watanabe

et al. 2019

Mol Med

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Background: There are two isoforms of sphingomyelin synthase (SMS): SMS1 and SMS2. SMS1 is located in the Golgi apparatus only while SMS2 is located in both the plasma membrane and the Golgi apparatus. SMS1 and SMS2 act similarly to generate sphingomyelin (SM). We have undertaken the experiments reported here on SMS and osteoblast differentiation in order to better understand the role SMS plays in skeletal development. Methods: We analyzed the phenotype of a conditional knockout mouse, which was generated by mating a Sp7 promoter-driven Cre-expressing mouse with an SMS1-floxed SMS2-deficient mouse (Sp7-Cre;SMS1 f/f ;SMS2 −/− mouse). Results: When we compared Sp7-Cre;SMS1 f/f ;SMS2 −/− mice with C57BL/6, SMS2-deficient mice (SMS1 f/f ;SMS2 −/−) and SP7-Cre positive control mice (Sp7-Cre, Sp7-Cre;SMS1 +/+ ;SMS2 +/− and Sp7-Cre;SMS1 +/+ ;SMS2 −/−), we found that although cartilage formation is normal, Sp7-Cre;SMS1 f/f ;SMS2 −/− mice showed reduced trabecular and cortical bone mass, had lower bone mineral density, and had a slower mineral apposition rate than control mice. Next, we have used a tamoxifen-inducible knockout system in vitro to show that SMS1 plays an important role in osteoblast differentiation. We cultured osteoblasts derived from ERT2-Cre;SMS1 f/f SMS2 −/− mice. We observed impaired differentiation of these cells in response to Smad1/5/8 and p38 that were induced by bone morphogenic protein 2 (BMP2). However, Erk1/2 phosphorylation was unaffected by inactivation of SMS1. Conclusions: These findings provide the first genetic evidence that SMS1 plays a role in bone development by regulating osteoblast development in cooperation with BMP2 signaling. Thus, SMS1 acts as an endogenous signaling component necessary for bone formation.

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RNA isolation from alveolar bone and gene expression analysis of RANK, RANKL and OPG: A new tool to monitor bone remodeling and healing in different bone substitutes used for prosthetic rehabilitation

Cited by 11 publications

References 14 publications

Demethylbelamcandaquinone B (Dmcq B) Is the Active Compound of Marantodes pumilum var. alata (Blume) Kuntze with Osteoanabolic Activities

Demethylbelamcandaquinone B (Dmcq B) Is the Active Compound of Marantodes pumilum var. alata (Blume) Kuntze with Osteoanabolic Activities

Cinnamaldehyde, a Promising Natural Preservative Against Aspergillus flavus

Deficiency of sphingomyelin synthase 1 but not sphingomyelin synthase 2 reduces bone formation due to impaired osteoblast differentiation

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