• 생명과학회지
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• 제27권5호
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• pp.501-508
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• 2017

Bone morphogenetic proteins (BMPs)는 다양한 세포기능을 조절하는 중요한 사이토카인 중 하나이다. 최근 BMP와 일주기 유전자들이 연관되어 있다는 연구결과들이 보고되고 있지만 조골세포에서 일주기 유전자인 Per1의 역할은 아직 명확하지 않다. 본 연구에서는 조골세포 분화에서 Per1의 역할을 조사하였다. MC3T3-E1 세포에서 BMP2 처리에 의해 Per1 mRNA 발현과 luciferase 활성이 증가하는 것을 확인하였다. 또한 Per1 과발현 실험을 통해서 Per1 유전자가 Runx2, ALP, OC의 발현을 증가시켰으며 ascorbic acid와 ${\beta}$-glycerophosphate에 의한 ALP 염색과 석회화가 Per1 과발현에 의해 더욱 증가하는 것을 확인하였다. 이상의 결과는 일주기 리듬을 조절하는 Per1 유전자가 조골세포의 분화를 촉진하는 인자로 작용함을 시사한다.

• 대한한방부인과학회지
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• 제25권2호
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• pp.23-42
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• 2012

Objectives: This study was performed to evaluate the effect of Samkieumgamibang (SKG) on osteoporosis. Methods: The osteoclastogenesis and gene expression were determined in RANKL-stimulated RAW 264.7 cell. And, osteoblastogenesis was also determined in rat calvarial cell. Results: SKG decreased the number of TRAP positive cell in osteoclast. It also decreased the expression of Cathepsin K, MMP-9, TRAP, c-fos, NAFTc1 and JNK1 in osteoclast. SKG increased the expression of iNOS in RANKL-stimulated in osteoclast. Otherwise, SKG inhibited TRAP activity in osteoclast. SKG increased cell proliferation, ALP activity, bone martix protein, collagen and nodule in osteoblast. Conclusions: It is concluded that SKG might decrease the bone resorption resulted from decrease of osteoclast differentiation and it's related gene expression. And, SKG might increase the bone formation resulted from increase of osteoblast function.

• BMB Reports
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• 제48권11호
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• pp.636-641
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• 2015

There are controversial findings regarding the roles of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway on bone metabolism under oxidative stress. We investigated how Nrf2/HO-1 pathway affects osteoblast differentiation of MC3T3-E1 cells in response to hydrogen peroxide (H₂O₂), N-acetyl cysteine (NAC), or both. Exposing the cells to H₂O₂ decreased the alkaline phosphatase activity, calcium accumulation, and expression of osteoblast markers, such as osteocalcin and runt-related transcription factor-2. In contrast, H₂O₂ treatment increased the expression of Nrf2 and HO-1 in the cells. Treatment with hemin, a chemical HO-1 inducer, mimicked the inhibitory effect of H₂O₂ on osteoblast differentiation by increasing the HO-1 expression and decreasing the osteogenic marker genes. Pretreatment with NAC restored all changes induced by H₂O₂ to near normal levels in the cells. Collectively, our findings suggest that H₂O₂-mediated activation of Nrf2/HO-1 pathway negatively regulates the osteoblast differentiation, which is inhibited by NAC.

• 생약학회지
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• 제45권4호
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• pp.326-331
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• 2014

In this study, we investigated pharmacologic activity of rice bran ethyl acetate fraction (RBE), based on their osteoblast enhancing effects. It has been found that REB have a stimulatory effect on the commitment of bi-potential mesenchymal precursor C2C12 cells into osteoblasts in the presence of BMP-2. Furthermore, RBE enhanced the BMP-2-stimulated induction of ALP, an early phase biomarker of osteoblast differentiation. In addition, Western blot analysis showed RBE enhanced the BMP-2-stimulated phosphorylation of p38, but not those of ERK or JNK. These findings show RBE has the potential to enhance the BMP-2-mediated commitment of C2C12 cells into osteoblasts and their differentiation through p38 activation.

• 대한한방부인과학회지
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• 제29권2호
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• pp.66-82
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• 2016

Objectives : This study was conducted to evaluate the effect of Kanghwalsokdan-tang Gamibang water extract (KSG) on osteoporosis. Methods : RANKL-stimulated RAW 264.7 was used to evaluate inhibitory effect of KSG osteoclast differentiation and gene expression. We counted TRAP (+) multinucleated cells and measured TRAP activity and mRNA expressions of osteoclastogenesis-related genes (NFATc1, MITF, JNK1, cathepsin K, MMP-9) to figure out the effect of KSG on osteoclast. Osteoblastogenesis was also determined in rat calvarial cell. Alkaline phosphatase (ALP) activity, bone matrix protein and collagen synthesis were measured by using murine calvarial cell. Results : KSG inhibited the differentiation of osteoclast precursor cell and expression of genes related osteoclastogenesis like NAFTc1, MITF, c-fos, JNK1, Cathepsin K, MMP-9 and TRAP. KSG increased cell division and function of osteoblast separated from the skull of a rat and ALP synthesis, biosynthesis of bone matrix protein and collagen. Conclusions : Reviewing these results, KSG has efficacy on osteoclast inhibition and osteoblast activation. After further study, KSG will be able to apply for osteoporosis treatment and prevention.

• 대한한의학방제학회지
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• 제19권2호
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• pp.61-71
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• 2011

Objectives : Kyungok-go(KOG), the first herbal formulation of donguibogam, has been used for treating of many symptoms of yin deficiency. In this study, we examined the effect of KOG on bone resorption. Methods : We determined the effects of water extract of KOG in RANKL(Receptor Activator for Nuclear Factor ${\kappa}B$ Ligand)-induced osteoclast differentiation culture system and osteoblast proliferation. In addition, we determined the effects of water extract of ABR on LPS-induced bone-loss with mice. Results : Water extract of KOG showed proliferation effect on osteoblast without cytotoxicity and no effect on RANKL-treated osteoclast differentiation. KOG rescued bone erosion by LPS induction in vivo study. Conclusions : These results demonstrated that KOG can be a useful remedy for treating of bone-loss disease such as osteoporosis.

• 한국식품영양과학회지
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• 제46권3호
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• pp.314-319
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• 2017

본 연구에서는 마가목 열매에서 추출한 chlorogenic acid의 유사체인 cryptochlorogenic acid(CCA)가 조골세포 분화에 미치는 영향에 대해서 알아보았다. 먼저 세포독성 여부를 확인하기 위해 MTT assay를 수행하였고 독성이 없다고 확인된 $5{\mu}M$의 농도에서 실험을 진행하였다. 그리고 조골세포로 분화할 수 있는 다분화능 세포인 C3H10T1/2와 조골세포인 MC3T3-E1에 CCA를 처리하여 표지 유전자인 Id1, Dlx5, Runx2의 발현을 확인하였다. 확인한 결과 표지유전자들의 발현이 대조군에 비교해서 증가한 것을 확인하였고, 그중 조골세포의 핵심 전사조절인자인 Runx2의 전사활성에 미치는 영향을 알아보기 위해 promoter assay를 수행하여 Runx2의 전사활성이 증가하는 것을 재확인하였다. 이러한 결과들을 토대로 CCA는 조골세포 분화를 촉진한다는 것을 알게 되었고, 골 질환 관련 제제로 CCA가 이용 가능할 수 있다고 생각된다.

• Journal of Dental Anesthesia and Pain Medicine
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• 제19권2호
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• pp.91-99
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• 2019

Background: The imbalance between osteoblasts and osteoclasts can lead to pathological conditions such as osteoporosis. It has been reported that opioid adversely affect the skeletal system, but it is inconsistent. Remifentanil is currently used as an adjuvant analgesic drug in general anesthesia and sedation. The aim of the present study was to investigate the effect of remifentanil on the osteoblast differentiation and mechanism involved in this effect. Methods: The C2C12 cells (mouse pluripotent mesenchymal cell line) were used as preosteoblast. Osteoblastic differentiation potency was determined by alkaline phosphatase (ALP) staining. C2C12 cell migration by remifentanil was evaluated using Boyden chamber migration assay. The expression of Runx2 and osterix was evaluated by RT-PCT and western blot analysis to investigate the mechanism involved in remifentanil-mediated osteoblast differentiation. Results: ALP staining showed that remifentanil increased significantly osteoblast differentiation. In Boyden chamber migration assay, C2C12 cell migration was increased by remifentanil. RT-PCR and western blot analysis showed that the expression of Runx2 and osterix was upregulated by remifentanil. Conclusions: We demonstrated that remifentanil increased osteoblast differentiation in vitro by upregulation of Runx2 and osterix expression. Therefore, remifentanil has the potential for assisting with bone formation and bone healing.

• Natural Product Sciences
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• 제24권4호
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• pp.235-240
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• 2018

Betula platyphylla var. japonica (Betulaceae), also known as Asian white birch, is an endemic medicinal tree, the bark of which has been used in Chinese traditional medicine for the treatment of various inflammatory diseases. In our continuing search for bioactive compounds from Korean natural resources, a phytochemical investigation of the bark of B. platyphylla var. japonica led to the isolation of 7-oxo-${\beta}$-sitosterol (1) and soyacerebroside I (2) from its ethanol extract as main components by liquid chromatography (LC)/mass spectrometry (MS)-based analysis. The structures of isolates were identified by comparison of $^1H$ and $^{13}C$ nuclear magnetic resonance spectroscopic data and physical data with the previously reported values and LC/MS analyses. To the best of our knowledge, this is the first study to demonstrate that the isolated compounds, 7-oxo-${\beta}$-sitosterol and soyacerebroside I, were isolated in B. platyphylla var. japonica. We examined the effects of the isolates on the regulation of adipocytes and osteoblast differentiation. These isolates (1 and 2) produced fewer lipid droplets compared to the untreated negative control in Oil Red O staining of the mouse mesenchymal stem cell line without altering the amount of alkaline phosphatase staining. The results demonstrated that both compounds showed marginal inhibitory effects on adipocyte differentiation but did not affect osteoblast differentiation.

• Journal of Periodontal and Implant Science
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• 제35권4호
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• pp.851-861
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• 2005

The nature of the implant surface can directly influence cellular response, ultimately affecting the rate and quality of new bone tissue formation. The aim of this in vitro study was to investigate if human osteoblast-like cells, Saos-2, would respond differently when plated on disks of magnesium titanate and machined titanium. Magnesium titanate disks were prepared using Micro Arc Oxidation(MAO) methods. Control samples were machined commercially pure titanium disks. The cell adhesion, proliferation and differentiation were evaluated by measuring cell number, and alkaline phosphatase(ALPase) activity at 1 day and 6 day after plating on the titanium disks. Measurement of cell number and ALPase activity in Saos-2 cells at 1 day did not demonstrate any difference between machined titanium and magnesium titanate. When compared to machined titanium disks, the number of cells was reduced on the magnesium titanate disks at 6 day, while ALPase activity was more pronounced on the magnesium titanate. Enhanced differentiation of cells grown on magnesium titanate samples was indicated by decreased cell proliferation and increased ALPase activity.