7-HYB, a Phenolic Compound Isolated from Myristica fragrans Houtt Increases Cell Migration, Osteoblast Differentiation, and Mineralization through BMP2 and β-catenin Signaling

The seeds (nutmegs) of Myristica fragrans Houtt have been used as popular spices and traditional medicine to treat a variety of diseases. A phenolic compound, ((7S)-8′-(benzo[3′,4′]dioxol-1′-yl)-7-hydroxypropyl)benzene-2,4-diol (7-HYB) was isolated from the seeds of M. fragrans. This study aimed to investigate the anabolic effects of 7-HYB in osteogenesis and bone mineralization. In the present study, 7-HYB promotes the early and late differentiation of MC3T3-E1 preosteoblasts. 7-HYB also elevated cell migration rate during differentiation of the preosteoblasts with the increased phosphorylation of mitogen-activated protein kinases (MAPKs) including ERK1/2, p38, and JNK. In addition, 7-HYB induced the protein level of BMP2, the phosphorylation of Smad1/5/8, and the expression of RUNX2. 7-HYB also inhibited GSK3β and subsequently increased the level of β-catenin. However, in bone marrow macrophages (BMMs), 7-HYB has no biological effects in cell viability, TRAP-positive multinuclear osteoclasts, and gene expression (c-Fos and NF-ATc1) in receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis. Our findings suggest that 7-HYB plays an important role in osteoblast differentiation through the BMP2 and β-catenin signaling pathway. It also indicates that 7-HYB might have a therapeutic effect for the treatment of bone diseases such as osteoporosis and periodontitis.

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TMARg, a Novel Anthraquinone Isolated from Rubia cordifolia Nakai, Increases Osteogenesis and Mineralization through BMP2 and β-Catenin Signaling

International Journal of Molecular Sciences ◽

10.3390/ijms21155332 ◽

2020 ◽

Vol 21 (15) ◽

pp. 5332

Author(s):

Kyung-Ran Park ◽

Joon Yeop Lee ◽

Bo-Mi Kim ◽

Sang Wook Kang ◽

Hyung-Mun Yun

Keyword(s):

Cell Migration ◽

Signaling Pathways ◽

Intracellular Signaling ◽

Osteoblast Differentiation ◽

Biological Effects ◽

Bone Diseases ◽

C2c12 Cells ◽

Alizarin Red ◽

Rubia Cordifolia ◽

Mitogen Activated Protein

Background: Plant extracts have long been regarded as useful medicines in the treatment of human diseases. Rubia cordifolia Nakai has been used as a traditional medicine, as it has pharmacological properties such as antioxidant and anti-inflammatory activity. However, the biological functions of TMARg, isolated from the roots of R. cordifolia, in osteoblast differentiation remain unknown. This study was performed to investigate the pharmacological effects and intracellular signaling of TMARg in the osteoblast differentiation of pre-osteoblast MC3T3-E1 cells and mesenchymal precursor C2C12 cells. Methods: Cell viability was evaluated using an MTT assay. Early and late osteoblast differentiation was examined by analyzing the activity of alkaline phosphatase (ALP), and by staining it with Alizarin red S (ARS). Cell migration was determined by using migration assays. Western blot analysis and immunocytochemical analysis were used to examine the intracellular signaling pathways and differentiation proteins. Results: In the present study, TMARg showed no cytotoxicity and increased the osteoblast differentiation in pre-osteoblasts, as assessed from the alkaline phosphate (ALP) staining and activity and ARS staining. TMARg also induced BMP2 expression and increased the p-smad1/5/8-RUNX2 and β-catenin pathways in both MC3T3-E1 and C2C12 cells. Furthermore, TMARg activated mitogen-activated protein kinases (MAPKs) and increased the cell migration rate. In addition, the TMARg-mediated osteoblast differentiation was suppressed by BMP and Wnt inhibitors with the downregulation of BMP2 expression. Conclusion: These findings demonstrate that TMARg exerts pharmacological and biological effects on osteoblast differentiation through the activation of BMP2 and β-catenin signaling pathways, and suggest that TMARg might be a potential phytomedicine for the treatment of bone diseases.

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Ziyuglycoside I Upregulates RUNX2 through ERK1/2 in Promoting Osteoblast Differentiation and Bone Mineralization

The American Journal of Chinese Medicine ◽

10.1142/s0192415x21500427 ◽

2021 ◽

pp. 1-18

Author(s):

Kyung-Ran Park ◽

Joon Yeop Lee ◽

MyoungLae Cho ◽

Hyung-Mun Yun

Keyword(s):

Cell Migration ◽

Osteoblast Differentiation ◽

Bone Mineralization ◽

Biological Effects ◽

Bone Diseases ◽

Nuclear Accumulation ◽

Herbaceous Plant ◽

Mrna Levels ◽

Dependent Manner ◽

Alizarin Red

Sanguisorba officinalis L. (Rosaceae) is a perennial herbaceous plant and its roots have been used as an important traditional medicine for over 2000 years. Ziyuglycoside I (Ziyu), an active compound isolated from the roots of S. officinalis L., has shown biological effects such as anti-oxidant, antiviral, and antiwrinkle activities. This study aimed to elucidate the underlying mechanisms of action of Ziyu on cytotoxicity, migration, and differentiation of pre-osteoblasts. Herein, at concentrations ranging from 1 to 100 [Formula: see text]M, Ziyu was not cytotoxic against pre-osteoblasts. Alkaline phosphatase activity assay and staining, and migration assay showed that Ziyu increased cell migration and promoted early osteoblast differentiation, followed by the enhancement of mineralized nodule formation in a dose-dependent manner, as indicated by Alizarin Red S staining. In addition, Ziyu increased the protein levels of runt-related transcription factor 2 (RUNX2) during osteoblast differentiation, whereas it did not affect the phosphorylation of Smad1/5/8 and GSK3b and expression of [Formula: see text]-catenin. Ziyu also activated ERK1/2 and mitogen-activated protein kinase during osteoblast differentiation, and ERK1/2 inhibitor attenuated Ziyu-mediated RUNX2 expression and nuclear accumulation. Furthermore, Ziyu-mediated early and late osteoblast differentiation was significantly suppressed by the inhibition of ERK1/2, which was accompanied by attenuation in the mRNA levels of osteoblast-related genes including bone sialoprotein, osteopontin, and osteocalcin. Taken together, the findings of this study provide evidence that Ziyu promotes cell migration, osteoblast differentiation, and bone mineralization and suggest a potential role for Ziyu in the treatment of bone diseases.

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Osteogenic Effects of Triterpene Saponin Soyasapogenol B on Differentiation, Mineralization, Autophagy, and Necroptosis

10.21203/rs.3.rs-1182818/v1 ◽

2021 ◽

Author(s):

Kyung-Ran Park ◽

Joon Yeop Lee ◽

Soo Hyun Kim ◽

Il Keun Kwon ◽

Hyung-Mun Yun

Keyword(s):

Cell Proliferation ◽

Osteoblast Differentiation ◽

Biological Effects ◽

Bone Diseases ◽

Adhesion Assay ◽

Pharmacological Properties ◽

Alizarin Red ◽

Triterpenoid Saponins ◽

Triterpene Saponin ◽

Osteogenic Activity

Abstract Background: Triterpenoid saponins are a diverse group of natural compounds in plants. A triterpene saponin, Soyasapogenol B (SoyB), from Arachis hypogaea (peanut) has various pharmacological properties. This study aimed to elucidate pharmacological properties and mechanisms of SoyB on bone-forming cells. Methods: Cell viability adhesion, and migration were analyzed using MTT assay, cell adhesion assay, and Boyden chamber assay. Osteogenic activity and osteogenicity were analyzed using alkaline phosphatase (ALP) staining and activity, and Alizarin Red S (ARS) staining. Cell signaling, protein expression, and autophagy were analyzed using Western blot analysis, immunofluorescence assay, and DAPGreen autophagy detection assay. Results and Conclusion: In the present study, SoyB (> 99.99% purity), triterpene saponin, was isolated from the fruit of A. hypogaea. At concentrations ranging from 1 to 20 mM, SoyB showed no cell proliferation effects, whereas 30 - 100 mM SoyB increased cell proliferation in MC3T3-E1 cells. Next, osteoblast differentiation was analyzed and found that SoyB enhanced ALP staining and activity and bone mineralization as evidence for early and late osteoblast differentiation. SoyB also induced RUNX2 expression in nucleus with the increased phosphorylation of Smad1/5/8 and JNK2 during osteoblast differentiation. In addition, SoyB-mediated osteoblast differentiation was not associated with autophagy and necroptosis. Furthermore, SoyB increased cell migration and adhesion with the upregulation of MMP13 levels during osteoblast differentiation. The findings of this study provide new evidence that SoyB possesses biological effects on osteogenic activity and osteogenicity in bone-forming cells, and suggest a potentially beneficial role for peanuts foods and drugs containing SoyB in the treatment and prevention of bone diseases.

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Biological Effects of Anti-RANKL Antibody and Zoledronic Acid on Growth and Tooth Eruption in Growing Mice

Scientific Reports ◽

10.1038/s41598-019-56151-1 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 3

Author(s):

Motoki Isawa ◽

Akiko Karakawa ◽

Nobuhiro Sakai ◽

Saki Nishina ◽

Miku Kuritani ◽

...

Keyword(s):

Zoledronic Acid ◽

Osteoblast Differentiation ◽

Pediatric Patients ◽

Root Formation ◽

Biological Effects ◽

Normal Growth ◽

Bone Diseases ◽

Bone Volume ◽

Tooth Eruption ◽

Tooth Root

AbstractThe anti-bone resorptive drugs denosumab, an anti-human-RANKL antibody, and zoledronic acid (ZOL), a nitrogen-containing bisphosphonate, have recently been applied for treatment of pediatric patients with bone diseases, though details regarding their effects in growing children have yet to be fully elucidated. In the present study, we administered these anti-resorptive drugs to mice from the age of 1 week and continued once-weekly injections for a total of 7 times. Mice that received the anti-RANKL antibody displayed normal growth and tooth eruption, though osteopetrotic bone volume gain in long and alveolar bones was noted, while there were nearly no osteoclasts and a normal of number osteoblasts observed. In contrast, ZOL significantly delayed body growth, tooth root formation, and tooth eruption, with increased osteoclast and decreased osteoblast numbers. These findings suggest regulation of tooth eruption via osteoblast differentiation by some types of anti-resorptive drugs.

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Phytochemical and Pharmacological Activity of Myristica fragrans Houtt (Myristicaceae)

INTERNATIONAL JOURNAL OF TOXICOLOGICAL AND PHARMACOLOGICAL RESEARCH ◽

10.25258/ijtpr.v9i01.9038 ◽

2017 ◽

Vol 9 (01) ◽

Author(s):

Md Arifur Rahman Chowdhury ◽

Manirujjaman . ◽

Md Mazedul Haq

Keyword(s):

Acetic Acid ◽

Phenolic Content ◽

Methanolic Extract ◽

Radical Scavenging ◽

Antioxidant Property ◽

Free Radical Scavenging ◽

Total Phenolic ◽

Writhing Test ◽

Myristica Fragrans ◽

Myristica Fragrans Houtt

Objective: Myristica fragrans Houtt commonly used as traditional medicine for alleviating of various disorders. The purpose of our study was to map out the in vitro antioxidant property and in vivo anti-hyperglycemic and analgesic effect of the methanolic extract of Myristica fragrans Houtt. (Seed and mace) (Myristicaceae) on Swiss albino mice. Methods: The processed powder of Myristica fragrans Houtt (seed and mace) were subjected to methanolic extraction by soxhlet filtration methods, and the desiccated extract was used for screening of antioxidant by DPPH free radical scavenging assessment as well as total phenolic content by using folin-ciocalteu reagent.Anti-hyperglycemic effect and analgesic action tested through alloxan induced antidiabetics test and acetic acid-tempted writhing test on mice. Results: In DPPH free radical scavenging assessment, free radicals neutralization expressed as % of inhibition 49.69±0.06% also by IC50 values as 68.43 µg/ml surmise middle level of antioxidant property. The total phenolic content expressed as 186.25 mg/g equivalent of gallic acid indicates, active phenolic content. Oral administration of 200 and 400 mg/kg of extract dose and reference drug vildagliptin (50 mg/kg) for the duration of the 4-day study period, and initiated % of inhibition the blood glucose level measured as 22.48%, 44.78% and 62.02% regard as the significant anti-hyperglycemic properties. The analgesic activity was investigated by using the acetic acid-induced writhing test in mice, at the dose of 200 mg/kg body and 400mg/kg weight, and resulting 50.4% and 68.10% correspondingly, which was considerably significant with a standard drug. Conclusion: The present study suggests that methanolic extract of seed and mace of Myristica fragrans Houtt can manage moderate oxidative stress as well as perform the painkilling action. Besides, prolong medication may enhance the new dimension of anti-hyperglycemic activity.

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A Photoactivable Formaldehyde Donor with Fluorescence Monitoring Reveals Threshold to Arrest Cell Migration

10.26434/chemrxiv.11388321.v1 ◽

2019 ◽

Author(s):

Lukas P Smaga ◽

Nicholas W Pino ◽

Gabriela E Ibarra ◽

Vishnu Krishnamurthy ◽

Jefferson Chan

Keyword(s):

Wound Healing ◽

Cell Migration ◽

Fluorescence Enhancement ◽

Biological Effects ◽

Cellular Systems ◽

Live Cells ◽

First Report ◽

Cell Lysate ◽

Intracellular Release ◽

Biological Analytes

Controlled light-mediated delivery of biological analytes enables the investigation of highly reactivity molecules within cellular systems. As many biological effects are concentration dependent, it is critical to determine the location, time, and quantity of analyte donation. In this work, we have developed the first photoactivatable donor for formaldehyde (FA). Our optimized photoactivatable donor, photoFAD-3, is equipped with a fluorescence readout that enables monitoring of FA release with a concomitant 139-fold fluorescence enhancement. Tuning of photostability and cellular retention enabled quantification of intracellular FA release through cell lysate calibration. Application of photoFAD-3 uncovered the concentration range necessary for arresting wound healing in live cells. This marks the first report where a photoactivatable donor for any analyte has been used to quantify intracellular release.

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Karekteristik Mikrokapsul Oleoresin Fuli Pala (Myristica Fragrans Houtt) Yang Dienkapsulasi Dengan Gum Arab

Jurnal Teknologi Pangan dan Hasil Pertanian ◽

10.26623/jtphp.v14i2.1281 ◽

2020 ◽

Vol 14 (2) ◽

pp. 1

Author(s):

Alston Millan

Keyword(s):

Scavenging Activity ◽

Myristica Fragrans ◽

Myristica Fragrans Houtt

ABSTRAK Tujuan penelitian ini adalah untuk mengetahui pengaruh formulasi berbagai total padatan yang terdiri dari gum Arab sebagai enkapsulan dan oleoresin fuli pala sebagai core terhadap karakteristik mikrokapsul yang dihasilkan. Variasi formulasi oleoresin fuli pala dan gum arab yang dicobakan adalah A1 (5%), A2 (10%), A3 (15%), A4 (20%) dan A5 (20%). Pembentukan emulsi dilakukan dengan Turrax homogenizer dengan kecepatan 4000 rpm selama 5 menit. Pembentukan mikrokapsul dilakukan dengan menggunakan pengering semprot (spray dryer) dengan suhu inlet dan outlet masing-masing 1100C dan 620C. Mikrokapsul yang dihasilkan dianalisis karakteristiknya yang meliputi rendemen mikrokapsul, kadar oleoresin terkapsulkan, kadar oleoresin tak terkapsulkan, kadar air, engel of repose, wettability dan rehidrasi, aktivitas antioksidan dengan metode RSA (radikal scavenging activity) DPPH (2-2Dhpypenil-2 Picrylhydrazil) dan kromatografi gas-spektrometri massa (GC-MS). Mikrokapsul yang terbaik diperoleh dengan variasi formulasi oleoresin fuli pala dan gum arab adalah A1 (5%). Sifat mikrokapsul yang diperoleh menpunyai rendemen mikrokapsul 24,68%, oleoresin terkapsulkan 92,68%, oleoresin tak terkapsulkan 7,607%, kadar air 8,444%, angel of repose 19,083 (0), wettability 5,9 (menit), aktivitas antioksidan (IC50) 1032 ppm dan kromatografi gas-spektrometri massa (GC-MS) dengan komponen terbesar penyusun mikrokapsul oleoresin fuli pala yaitu sabinene hexane, 4-methylene-1-, myristycin, phenol 2,6 dimetoksi-4-2 (2-peopenil), phenol, 2-metoksi-4-(propennyl).

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Zinc Sulfate Stimulates Osteogenic Phenotypes in Periosteum-Derived Cells and Co-Cultures of Periosteum-Derived Cells and THP-1 Cells

Life ◽

10.3390/life11050410 ◽

2021 ◽

Vol 11 (5) ◽

pp. 410

Author(s):

Jin-Ho Park ◽

Su A Park ◽

Young-Hoon Kang ◽

So Myeong Hwa ◽

Eun-Byeol Koh ◽

...

Keyword(s):

Bone Resorption ◽

Zinc Sulfate ◽

Nuclear Factor Kappa B ◽

Osteoblastic Differentiation ◽

Nuclear Factor ◽

Mitogen Activated Protein Kinases ◽

Extracellular Signaling ◽

Receptor Activator ◽

Mitogen Activated Protein ◽

Osteoclastic Differentiation

Coupling between osteoblast-mediated bone formation and osteoclast-mediated bone resorption maintains both mechanical integrity and mineral homeostasis. Zinc is required for the formation, mineralization, growth, and maintenance of bones. We examined the effects of zinc sulfate on osteoblastic differentiation of human periosteum-derived cells (hPDCs) and osteoclastic differentiation of THP-1 cells. Zinc sulfate enhanced the osteoblastic differentiation of hPDCs; however, it did not affect the osteoclastic differentiation of THP-1 cells. The levels of extracellular signaling-related kinase (ERK) were strongly increased during osteoblastic differentiation in zinc sulfate-treated hPDCs, compared with other mitogen-activated protein kinases (MAPKs). Zinc sulfate also promoted osteogenesis in hPDCs and THP-1 cells co-cultured with the ratio of one osteoclast to one osteoblast, as indicated by alkaline phosphatase levels, mineralization, and cellular calcium contents. In addition, the receptor activator of nuclear factor kappa B ligand (RANKL)/osteoprotegerin (OPG) ratio was decreased in the zinc sulfate-treated co-cultures. Our results suggest that zinc sulfate enhances osteogenesis directly by promoting osteoblastic differentiation and osteogenic activities in osteoblasts and indirectly by inhibiting osteoclastic bone resorption through a reduced RANKL/OPG ratio in co-cultured osteoblasts and osteoclasts.

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