• Preventive Nutrition and Food Science
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• 제16권4호
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• pp.291-298
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• 2011

Yam extracts (Dioscorea batatas) have been reported to possess a variety of functions. However, studies on its osteogenic properties are limited. In this study, we investigated the effect of ethanol and water extracts on osteoblast proliferation and bone matrix protein synthesis, type I collagen and alkaline phosphatase (ALP), using osteoblastic MC3T3-E1 cell model. MC3T3-E1 cells were cultured with yam ethanol and water extracts (0~30 mg/L) within 39 days of osteoblast differentiation period. Cell proliferation was measured by MTT assay. Bone matrix proteins were assessed by the accumulation of type I collagen and ALP activity by staining the cell layers for matrix staining. Also, the secreted (media) matrix protein concentration (type I collagen) and enzyme activity (ALP) were measured colorimetrically. Yam ethanol and water extracts stimulated cell proliferation within the range of 15~30 mg/L at 15 day treatment. The accumulation of type I collagen in the extracellular matrix, as well as secreted collagen in the media, increased with increasing doses of yam ethanol (3~15 mg/L) and water (3~30 mg/L) extracts. ALP activity was not affected by yam ethanol extracts. Our results demonstrated that yam extracts stimulated osteoblast proliferation and enhanced the accumulation of the collagenous bone matrix protein type I collagen in the extracellular matrix. These results suggest that yam extracts may be a potential activator for bone formation by increasing osteoblast proliferation and increasing bone matrix protein type I collagen. Before confirming the osteogenic action of yam, further studies for clarifying how and whereby yam extracts can stimulate this ostegenesis action are required.

• 대한의생명과학회지
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• 제11권3호
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• pp.319-326
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• 2005

It is well known that oxidative stress of reactive oxygen species (ROS) may be a causative factor in the pathenogenesis of bone disorder on osteoblast or osteoclast. The purpose of this study was to evaluate the cytotoxicity of oxidative stress, protective effect of glutamate receptor antagoinst against ROS-induced osteotoxicity, secretion of tumor necrosis factor $(TNF)-\alpha$ and the expression of c-fos gene in the cultured rat osteoblasts and osteoclasts. Cell viability by MTS assay or !NT assay, activity of glutathione peroxidase (GPx), lipid peroxidation (LPO) activity, protein synthesis by sulforhodamine B (SRB) assay, alkaline phosphatase (ALP) activity, lactate dehydrogenase (LDH) activity, MTS assay for NMDA (N-methyl-D-aspartate) receptor antagonist or AMPA/kainate receptor antagonist, measurement for $TNF-\alpha$, and c-fos gene expression were performed after these cells were treated with or without various cocentrations of xanthine oxidase (XO), hypoxanthine (HX), D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6,7-dinitroquinoxaline-2,3-dione (DNQX), respectively. In this study, XO/HX showed decreased cell viability and glutathione peroxidase (GPx) activity, but it showed increased LPO activity, $TNF-\alpha$ secretion and c-fos expression. APV and CKA incresed protein sythesis and ALP activity. While, CNQX or DNQX did not show any protective effect in LDH activity or cell viability. From these results, XO/HX showed cytotoxic effect in cultured rat osteoblast or osteoclast, and also NMDA receptor antagonist such as APV or CKA was effective in blocking XO/HX-induced osteotoxicity in these cultures.

• 대한임상검사과학회지
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• 제51권3호
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• pp.379-385
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• 2019

골다공증은 호르몬의 변화와 무기질 감소에 의해 골밀도의 감소를 유발하여 골절의 위험을 높이는 질환이다. 최근 보고에 의하면, 간염바이러스 및 에이즈 치료제로 사용되고 있는 Adefovir dipivoxil (ADV)의 장기적 복용에서 골다공증 부작용이 유발할 수 있음이 보고 되고 있다. 이에 대한 연구수행을 위해 골모세포주 hFOB1.19와 혈관내피세포 HUVEC을 이용하여 ADV에 대한 생물학적 연관성을 평가하였다. 우선적으로 ADV를 농도별로 처리한 후 각 세포와 핵의 형태학적 분석을 위해 DAPI와 crystal violet 염색을 시행하였다. 또한 세포 증식에 대한 약물 효과를 평가하기 위하여 CCK-8분석과 골모세포에 대한 분화유도 및 억제 효과를 확인하기 위하여, ALP 염색을 진행하였다. 그 결과, ADV는 hFOB1.19 세포와 HUVEC 세포에서 농도 의존적으로 세포의 비대 현상을 유발하였고, 세포의 증식이 억제되었다. 이러한 원인들을 규명하기 위해 TGF-${\beta}$발현을 조사하였을 뿐만 아니라 이러한 발현 감소에 의한 생물학적 영향이 골모세포로부터 골세포로의 분화 과정에 관여하고 있음을 확인 할 수 있었다. 결론적으로, 본 연구에서는 ADV 약물이 골모세포와 혈관내피세포의 TGF-${\beta}$의 발현을 억제하여 핵의 크기 증가와 세포형태의 비대증을 유발하며, 세포의 증식억제 및 골모세포 분화능에 영향을 줌으로서 골다공증을 유발할 수 있는 가능성을 확인하였다. 이러한 결과는 ADV 복용에 따른 골다공증 발병 원인을 이해하기 위한 기초 연구 및 이를 이용한 임상영역에 활용 될 수 있을 것으로 사료된다.

• 대한치과보철학회지
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• 제43권6호
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• pp.751-763
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• 2005

Statement of problem. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. The surface quality of the implant depends on the chemical, physical, mechanical and topographical properties of the surface. The different properties will interact with each other and a change in thickness of the oxide layer may also result in a change in surface energy, the surface topography and surface, chemical composition. However, there is limited the comprehensive study with regard to changed surface and biologic behavior of osteoblast by anodization. Purpose of study. The aim of this study was to analyze the characteristics of an oxide layer formed and to evaluate the cellular biologic behaviors on titanium by anodic oxidation (anodization) by cellular proliferation, differentiation, ECM formation and gene expression. And the phospholipase activity was measured on the anodized surface as preliminary study to understand how surface properties of Ti implant are transduced into downstream cellular events. Methods and Materials. The surface of a commercially pure titanium(Grade 2) was modified by anodic oxidation. The group 1 samples had a machined surface and other three experimental specimens were anodized under a constant voltage of 270 V(Group 2), 350 V(Group 3), and 450 V(Group 4). The specimen characteristics were inspected using the following five categories; the surface morphology, the surface roughness, the thickness of oxide layer, the crystallinity, and the chemical composition of the oxide layer. Cell numbers were taken as a marker for cell proliferation. While the expression of alkaline phosphatase and Runx2 (Cbfa1) was used as early differentiation marker for osteoblast. The type I collagen production was determined, which constitutes the main structural protein of the extracellular matrix. Phospholipase $A_2$ and D activity were detected. Results. (1) The anodized titanium had a porous oxide layer, and there was increase in both the size and number of pores with increasing anodizing voltage. (2) With increasing voltage, the surface roughness and thickness of the oxide film increased significantly (p<0.01), the $TiO_2$phase changed from anatase to rutile. During the anodic oxidization, Ca and P ions were more incorporated into the oxide layer. (3) The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity, type I collagen production, and Cbfa 1 gene expression increased significantly (p<0.01), while the cell proliferation decreased. (4) In preliminary study on the relation of surface property and phospholipase, PLD activity was increased but $PLA_2$ activity did not changed according to applied voltage. Conclusion. The anodized titanium shows improved surface characteristics than the machined titanium. The surface properties acquired by anodization appear to give rise more mature osteoblast characteristics and might result in increased bone growth, and contribute to the achievement of a tight fixation. The precise mechanism of surface property signaling is not known, may be related to phospholipase D.

• 한국식품영양과학회지
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• 제39권2호
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• pp.203-209
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• 2010

본 연구를 통하여 여주 추출물은 세포 증식을 제외하고는 조골세포에 긍정적인 영향을 미치지 못하였지만 머위 추출물은 세포의 증식, ALP 활성, bone nodule의 형성이 대조군과 비슷한 결과를 나타내거나 높은 경향을 나타냄으로써 조골세포의 골 형성 과정인 증식, 기질의 성숙, 기질의 석회화의 세 단계에서 유효성을 증명하였다. 또한 OPG mRNA의 2배 이상의 증가는 조골세포의 골 형성에 주요 매개 물질로서 가능성이 있음을 밝혔다. 따라서 머위 추출물은 골수의 미세 환경에서 세포의 조절작용을 하는 물질로 여겨지며, 골다공증을 포함한 각종 골 결손 질환의 예방과 치료약 개발에 긍정적인 가능성을 제시할 것이라 사료된다.

• 생명과학회지
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• 제16권6호
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• pp.925-931
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• 2006

골조직은 골아세포, 파골세포, 골세포 등으로 구성되며, 골개조시 여러 인자가 세포증식, 분화, 활성화 및 골대사 조절에 관여한다. 이때 조골세포의 활성은 골형성에 중요하므로, 본 연구에서는 MC3T3-E1 조골세포주를 이용하여 해조류로 조제한 알칼리의 ENA-A 이온수가 조골세포의 증식과 분화활성에 미치는 영향을 조사하였다. 먼저 ENA-A 이온수가 조골세포의 성장에 미치는 영향을 MTT 검색법으로 조사 한 결과, ENA-A 이온수 2% 처리시 대조군과 비교하여 134% 증가하여 조골세포에 대해 높은 성장률을 보였다. 또한 ALP 효소 활성에 미치는 영향을 9일간 배양하여 그 변화를 측정하였다. 결과, 농도 1, 2, 4, 8% 처리시 112, 150, 188%까지 ALP 활성을 증가시켰다. ENA-A 이온수는 다시 ALP 효소 염색법과 Alizarin Red 염색으로 조골세포의 ALP활성유도, 분화와 석회화 형성능을 재확인하였으며 골기질 유전자의 발현의 변화도 확인하였다. 이에 본 연구결과를 통해 각종 무기질 성분을 함유한 ENA 이온수가 조골세포의 분화시에 hydroxyapatite 형성에 영향을 줌으로써 골다공증 예방에 효과가 있을것으로 생각하여 이를 보고하는 바이다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권4호
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• pp.419-427
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• 2008

The present study aimed to investigate the osteogenic potentials of differentiated osteoblast-like cells (DOCs) induced from bone marrow-derived mesenchymal stem cells (MSCs) on ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) with recombinant human bone morphogenetic protein (rhBMP-2) in vitro. Osteoblast differentiation was induced in confluent cultures by adding 100 nM dexamethasone, 10 mM ${\beta}$-glycerophosphate, 50 mM L-ascorbic acid. The Alizarin red S staining and reverse transcriptase-polymerase chain reaction (RT-PCR) were perfomed to examine the mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), receptor activator for nuclear factor ${\kappa}B$ ligand (RANKL), runt-related transcription factor 2 (RUNX2), collagen-Ⅰ (COL-Ⅰ). There were no significant differences in the osteogenic potentials of DOCs induced from MSCs on ${\beta}-TCP(+/-)$. According to the incubation period, there were significant increasing of Alizadin red S staining in the induction 3 weeks. The mRNA expression of ALP, RUNX2, and RANKL were higher in DOCs/${\beta}-TCP(-)$ than DOCs/${\beta}-TCP(+)$. According to rhBMP-2 concentrations, the mRNA expression of BSP was significantly increased in DOCs/${\beta}-TCP(+)$ compared to that of DOCs/${\beta}-TCP(-)$ on rhBMP 10 ng/ml. Our study presented the ${\beta}-TCP$ will have the possibility that calcium phosphate directly affect the osteoblastic differentiation of the bone marrowderived MSCs.

• BMB Reports
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• 제46권11호
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• pp.527-532
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• 2013

Gangliosides are complex glycosphingolipids that are the major component of cytoplasmic cell membranes, and play a role in the control of biological processes. Human mesenchymal stem cells (hMSCs) have received considerable attention as alternative sources of adult stem cells because of their potential to differentiate into multiple cell lineages. In this study, we focus on various functional roles of gangliosides in the differentiation of hMSCs into osteoblasts or neuronal cells. A relationship between gangliosides and epidermal growth factor receptor (EGFR) activation during osteoblastic differentiation of hMSCs was observed, and the gangliosides may play a major role in the regulation of the differentiation. The roles of gangliosides in osteoblast differentiation are dependent on the origin of hMSCs. The reduction of ganglioside biosynthesis inhibited the neuronal differentiation of hMSCs during an early stage of the differentiation process, and the ganglioside expression can be used as a marker for the identification of neuronal differentiation from hMSCs.

• 한국식품조리과학회지
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• 제28권3호
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• pp.311-318
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• 2012

The correlation between osteoporosis and reactive oxygen species (ROS)-induced oxidative stress was investigated. Thus, interest in food and plants with antioxidant effects that can reduce damage caused by ROS during bone metabolism is heightening. In this study, the antioxidant effect of Taraxacum mongolicum on proliferation and differentiation of MC3T3-E1 cells under H2O2-induced oxidative stress was studied to investigate its protective effect against oxidative stress and its availability as an antioxidant material related to bone diseases. As a result, total polyphenol and total flavonoid contents of T. mongolicum were 33.65 mg/g and 4.45 mg/g, respectively. The T. mongolicum extract increased proliferation of both MC3T3-E1 cells and differentiated osteoblasts under $H_2O_2$-induced oxidative stress conditions. In addition, two differentiation markers, alkaline phosphatase activity and mineralization level in the T. mongolicum extract, tended to increase. These results indicate that T. mongolicum extract suppressed the damage to osteoblasts under oxidative stress and that it is potential antioxidant materials for preventing bone diseases.

• 치위생과학회지
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• 제15권4호
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• pp.488-494
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• 2015

치주인대세포의 효과적인 조절은 성공적인 치주 조직 재생에 중요한 역할을 한다. NFATc1의 활성화가 골모세포에서 분화를 자극하지만, 치주인대세포가 골모세포로 분화하는 과정에서 NFATc1의 역할은 아직 보고되지 않았다. 본 연구는 hPDLCs가 골모세포로 분화하는 동안 NFATc1의 mRNA의 발현과 단백질 발현이 유도됨을 처음으로 확인하였다. CsA에 의한 NFATc1의 억제는 세포증식을 감소시켰다. 게다가, CsA를 처리한 결과, 분화표지자, ALP activity 및 광화결정형성을 감소시켰다. 이러한 연구 결과는 NFATc1이 치주 재생을 위한 골모세포 분화에 중요한 조절자 역할을 할 수 있을 것으로 생각된다.