• Title/Summary/Keyword: HOS cell

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Biological Effects of Fibronectin Type III 10 domain on Human Osteoblast-like cells (Fibronectin type III 10 도메인이 조골양 세포에 미치는 생물학적 영향)

  • Lee, Chang-Seok;Jang, Jun-Hyeog;Kim, Tae-Il;Lee, Yong-Moo;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.293-301
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    • 2004
  • 1. 연구 목적 Fibronectin은 세포외기질의 주요성분인 거대 당단백질로서, 조골세포의 부착과 증식 및 이동능에 중요한 역할을 담당한다고 알려져 있다. 이러한 fibronectin의 조골세포에 대한 영향을 실제 임상에 적용하기 위해서, 전체 fibronectin 단백질을 사용하는 것은 면역학적으로나 경제적으로 많은 단점을 안고 있어서, 유효한 반응단위만을 추출하여 활용하는 것이 바람직한 방법으로 알려져 있다. 이 연구의 목적은 세포부착에 주로 관여하는 fibronectin type III분절 중 10번 도메인이 조골양 세포에 미치는 영향을 전체 fibronectin단백질과 fibronectin type III 7-10 도메인 분절과 비교, 관찰하는 것이다. 2. 연구 방법 사람의 fibronectin을 기초로 한 적절한 primer로서, 유전자 재조합법을 이용하여 fibronectin type III 10 도메인과 fibronectin type III 7-10 도메인 분절을 얻었으며, 전체 fibronectin분자는 상용으로 준비하여 24-well 세포배양 용기에 도포하였다. 배양된 조골양세포(HOS cell)를 $1x10^5$ cells/well의 농도로 각 well에 분주하여 $37^{\circ}C$에서 1시간 배양을 하였다. Cell adhesion assay를 실시하기 위해 10% formaldehyde로 고정시키고 1% Crystal Violet으로 염색하여 광학현미경을 관찰 후 2% SDS를 처리하여 microplate reader기를 이용하여 570nm에서 혼탁정도를 측정하였다. 음성대조군으로는 RPMI 용액을 사용하였다. 동일한 방법을 이용하여 준비된 $35mm^2배양접시에 HOS cell을 $37^{\circ}C$에서 4일간 배양 후, MTS assay를 이용하여 세포 증식도에 미치는 영향을 관찰하였다. 6일째 405nm에서 활성화된 세포에서 분비된 p-nitrophenol을 이용한 alkaline phosphatase activity를 측정하였다. 3. 결과 및 고찰 Fibronectin type III 10 도메인은 HOS cell에 대한 생물학적인 효과면에서, 전체 fibronectin 분자 및 fibronectin type III 7-10 분절과 통계적으로 유사한 세포부착도를 보여주었으며, 세포증식도와 alkaline phosphatse 활성도면에서도 큰 차이가 나타나지 않았다. 이상의 연구결과로 볼 때, fibronectin type III 10 도메인이 조골세포의 증식을 목적으로 사용하는 생체재료의 표면개질 부착물질로 응용할 수 있는 가능성이 있다고 하겠다.

Effects of Extracts from Fusobacterium nucleatum on the Growth of Human Gingival Fibroblasts and HOS 941 Cells, and on the TNF-α Production of Mouse Splenocytes (Fusobacterium nucleatum 추출물이 사람 치은 섬유아세포와 HOS 941세포의 성장과 마우스 비장세포의 TNF-α 생성에 미치는 효과)

  • Oh, Hee-Myung;Song, Yo-Han;Shin, Keum-Back
    • Journal of Oral Medicine and Pain
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    • v.24 no.4
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    • pp.361-374
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    • 1999
  • F. nucleatum is a gram-negative obligate anaerobe which is the principal and most frequent cause of gingival inflammation and is the predominant pathogen isolated in subsequent periodontal breakdown. It is also one of the most numerous bacteria found in subgingival plaque samples from healthy sites; its numbers are about 10-fold greater in plaque from periodontally diseased sites. The purpose of this study is to examine the effects of outer membrane(OM), outer membrane vesicle(OMV), and lipopolysaccharide(LPS) from F. nucleatum ATCC 25586 strain on the growth of human gingival fibroblasts and HOS 941 cells, and on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes. For the examination of cytotoxic effects, $TNF-{\alpha}$ production and $TNF-{\alpha}$ mRNA expression, the MTT assay, the ELISA and the RT-PCR were performed, respectively. All extracts of F. nucleatum tested were cytotoxic to both of human gingival fibroblasts and HOS 941 cells, and the significant difference of cytotoxic activity among the extracts was not observed. In the effects of these extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes, all extracts of F. nucleatum tested also stimulated the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression, but the effects of the OM extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression were higher than those of the OMV and the LPS extracts. The pattern of the $TNF-{\alpha}$ mRNA expression was similar to that of the $TNF-{\alpha}$ production. These results indicate that F. nucleatum seems to contribute to the pathogenesis of periodontal diseases at least by its cytotoxicity, directly and its $TNF-{\alpha}$ production, indirectly.

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The Effects of Boron on the Proliferation of Osteoblastic and Neuroblastoma Cells

  • Choi, Hye-Sook;Hang, Do;Choi, Mi-Kyeong;Lee, Sung-Ryul;Pyo, Suhkneung;Son, Eun-Wha;Kim, Mi-Hyun
    • Preventive Nutrition and Food Science
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    • v.10 no.4
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    • pp.353-356
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    • 2005
  • It has been recently reported that boron affects bone metabolism in humans and animals. In this study we examined whether boron affects the proliferation on various cell types, MG-63, HOS, Raw 264.7 and SK-N-SH. When treated with different concentrations of boron $(1,\;10,\;100{\mu}M)$ for 24 and 48 hr, the proliferation of MG-63 cells was enhanced at $10{\mu}M\;(p<0.05)$, for 24 hr. In HOS cells, boron had no effect on cell proliferation at 24 or 48 hr. In addition, treatment of pre-osteoclastic cells (Raw 264.7) with 1, 10, $100{\mu}M$ boron resulted in no effect on cell proliferation. Proliferation of neuronal cells (SK-N-SH) was enhanced by boron in a concentration dependent manner at low concentrations (0.1, 0.5, $1{\mu}M$). Besides proliferation activity, boron has an effect on the enhancement of NO production in SK-N-SH cells in a concentration-dependent manner. These studies showed that boron enhances proliferation of osteoblastic cells (especially MG-63), depending upon the concentration of boron. These results also provide further evidence of the positive effects of boron in neuronal disease.

ANALYSES OF THE EXPRESSION PROFILES OF GENES RESPONSIBLE FOR THE GROWTH OF OSTEOBLAST UPON VELVET ANTLERS TREATMENT (녹용이 조골세포(造骨細胞) 성장에 미치는 유전자 발현 profile 분석에 대한 연구)

  • Lee, Jong-Woo;Kim, Doeg-Kon
    • The Journal of Pediatrics of Korean Medicine
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    • v.16 no.1
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    • pp.39-74
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    • 2002
  • Recently, increased attention has been paid to the growth of the height of children and adolescents. To accelerate growth, velvet antlers are typically used in Oriental medicine. The present study investigated the effects of velvet antlers of velvet antlers on bone growth using the cell line of Human Osteosarcoma (Hos), derived from the bone-generating cells essential to bone growth. In order to give certain stress to this Hos, the medium contained 1% FBS was used for culturing for Hos cell instead of 10% in control. In this condition of which the proliferation had been significantly decreased, the ethanol extract of upper part of velvet antlers was added, As a result, the cells proliferation rate was significantly increased. Using Oligonucleotide DNA microarray, comparison and analyses were done to see what kind of specific genes would be differentially expressed. The result showed that as opposed to the control group, the stressed group indicated a decrease in the expressions of 6 kinds of genes such as, Id1, retinoid X receptor(RXRB) and 14-3-3 epsilon, etc. The velvet antler treated group, as opposed to the control group, showed a decreased in the expressions of 8 kinds of genes such as Id1, etc. and an increase in the expressions of 24 kinds of genes. The number of genes that showed differences in the velvet antler treated group compared with the stressed group was 7 the expression of 1 kind of gene was decreased, and the expressions of 6 kinds of genes were increased. Considering the mechanism by which velvet antlers affected the growth of osteoblast through reviewing the functions of these genes, the following results were attained. The constraint in the proliferation of Hos cells resulting from the medium contained 1% FBS seems to be caused by three important factors: 1) the decrease of the expression of 14-3-3 epsilon involved in the signal transduction and metabolism of growth, 2) the decrease of the expression of Id1 gene involved in the metabolism of bone formation, and 3) the decreased of expression of RXRB gene involved in the metabolism of retinoci acid. It is suggested that the improvement of the cell proliferating effects by velvet antler treatment, in stressed condition si mediated by increment of 6 genes particularly 14-3-3 epsilon, RXRB, and IGF2, with are the crucial factors for the cell growth and differentiation, metabolism of retinoic acid and osteoblast proliferation, respectively.

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Involvement of p53-Mediated Mitochondrial Stress in the Apoptosis Induced by Flavonoids Purified from Rhus verniciflua Stokes in Human Osteosarcoma Cells

  • Chung, Song-Woo;Lee, Seung-Ah;Park, Jong-Sun;Ryu, Kwon-Woo;Jang, Mun-Ju;Park, Song-Soo;Lee, Choon-Bong;Kim, Jong-Ghee;Jeon, Young-Mi;Lee, Jeong-Chae
    • Natural Product Sciences
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    • v.13 no.1
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    • pp.1-5
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    • 2007
  • Dietary flavonoids have antioxidant and antitumor promoting effects. Rhus verniciflua Stokes (RVS) is a flavonoid-rich herbal medicine and has long been used as a food additive and an antitumor agent in Korea. Previous study demonstrated that a purified flavonoid fraction prepared from RVS, herein named RCMF (the RVS chloroform-methanol fraction), exhibited growth inhibition and induced apoptosis in human osteosarcoma(HOS) cells. This study evaluated if p53-mediated pathway is associated with the RCMF-induced apoptosis in HOS cells. RCMF was shown to be capable of inducing apoptosis of the cells, as expected, and transparently increased p53 expression in the cells. However, the RCMF-induced cytotoxicity was suppressed by transfecting the cells with antisense p53 oligonucleotide, which also inhibited the decrease of Bcl-2 and the increase of Bax in mitochondria, and the release of cytochrome c into cytosol. This finding suggests that p53-mediated mitochondrial stress is required for RCMF-induced apoptosis in HOS cells.

STUDIES ON ANTICANCER EFFECTS OF EXTRACTS CAESALPINIA SAPPAN ON ORAL CARCINOMA AND OSTEOSARCOMA CELLS (소목 추출물의 구강암 및 골육종 세포주에 대한 항암작용에 관한 연구)

  • Lee, Jong-Su;Kim, Yeo-Gab;Kim, Jeong-Hee
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.4
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    • pp.281-288
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    • 2001
  • Anticancer effect of methanol extract of Caesalpinia sappan L. on oral carcinoma (KB) and osteosarcoma (HOS) cells were investigated in this study. In order to elucidate the anticancer mechanism of Caesalpinia sappan L, we analyzed telomerase inhibitory effect of the methanol extract of Caesalpinia sappan L. In addition we prepared 5 fraction samples according to its polarity differences and analyzed anticancer effects on oral carcinoma and osteosarcoma cells. Following results are obtained in this study. 1. 50% cell proliferation inhibitory value ($IC_{50}$) of the methanol extract of Caesalpinia sappan L. against oral carcinoma (KB) cells and osteosarcoma (HOS) cells were $9.0{\mu}g/ml$ and $10.9{\mu}g/ml$, respectively. 2. The methanol extract of Caesalpinia sappan L. showed inhibitory effect of telomerase which is required for cancer cell immortality. Therefore, it seems that the anticancer effect of methanol extract of Caesalpinia sappan is at least partially due to telomerase inhibitory effect. 3. Five fraction samples were prepared according to its polarity and 88.7% of ingredient of total methanol extract was transferred to ethylacetate fraction. Thin layer chromatography analysis showed that dichloromethane fraction contained ingredient with relatively high polarity and ethylacetate fraction contained similar ingredient found in total methanol extract. 4. Anticancer effect was observed in n-hexane, dichloromethane, and ethylacetate fractions. The highest anticancer effect was found in dichloromethane fraction which had $IC_{50}$ value of 4.4 and $>4.0{\mu}g/ml$ against oral carcinoma (KB) cells and osteosarcoma (HOS) cells, respectively.

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ANTICANCER EFFECTS OF CAESALPINIA SAPPAN EXTRACTS ON ORAL CARCINOMA AND OSTEOSARCOMA CELLS (소목 추출물의 구강암 및 골육종 세포주에 대한 항암작용에 관한 연구)

  • Lee, Jong-Su;Kim, Jeong-Hee;Kim, Yeo-Gab
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.6
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    • pp.583-590
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    • 2007
  • Anticancer effect of methanol extract of Caesalpinia sappan L. on oral carcinoma(KB) and osteosarcoma(HOS) cells were investigated in this study. In order to elucidate the anticancer mechanism of Caesalpinia sappan L, we analyzed telomerase inhibitory effect of the methanol extract of Caesalpinia sappan L. In addition, we prepared 5 fraction samples according to its polarity differences and analyzed anticancer effects on oral carcinoma and osteosarcoma cells. Following results are obtained in this study. 1. 50% cell proliferation inhibitory value($IC_{50}$) of the methanol extract of Caesalpinia sappan L. against oral carcinoma(KB) cells and osteosarco ma(HOS) cells were $9.0{\mu}g/ml\;and\;10.9{\mu}g/ml$, respectively. 2. The methanol extract of Caesalpinia sappan L. showed inhibitory effect of telomerase which is required for cancer cell immortality. Therefore, it seems that the anticancer effect of methanol extract of Caesalpinia sappan is at least partially due to telomerase inhibitory effect. 3. Five fraction samples were prepared according to its polarity and 88.7% of ingredient of total methanol extract was transferred to ethylacetate fraction. Thin layer chromatography analysis showed that dichloromethane fraction contained ingredient with relatively high polarity and ethylacetate fraction contained similar ingredient found in total methanol extract. 4. Anticancer effect was observed in n-hexane, dichloromethane, and ethylacetate fractions. The highest anticancer effect was found in dichloromethane fraction which had $IC_{50}$ value of $4.4{\mu}g/ml$ and > $4.0{\mu}g/ml$ against oral carcinoma(KB) cells and osteosarcoma(HOS) cells, respectively.

Handover based on Maximum Cell Residence Time and Adaptive TTT for LTE-R High-Speed Railways

  • Cho, Hanbyeog;Han, Donghyuk;Shin, Sungjin;Cho, Hyoungjun;Lee, Changsung;Lim, Goeun;Kang, Mingoo;Chung, Jong-Moon
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.11 no.8
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    • pp.4061-4076
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    • 2017
  • With the development of high-speed railway technologies, train velocities can now reach speeds up to 350 km/h, and higher in the future. In high-speed railway systems (HSRs), loss of communication can result in serious accidents, especially when the train is controlled through wireless communications. For to this reason, operators of Long Term Evolution for Railway (LTE-R) communication systems install eNodeBs (eNBs) with high density to achieve highly reliable communications. However, densely located eNBs can result in unnecessary frequent handovers (HOs) resulting in instability because, during every HO process, there is a period of time in which the communication link is disconnected. To solve this problem, in this paper, an HO scheme based on the maximum cell residence time (CRT) and adaptive time to trigger (aTTT), which are collectively called CaT, is proposed to reduce unnecessary HOs (using CRT estimations) and decrease HO failures by improving the handover command transmission point (HCTP) in LTE-R HSR communications.

Wettability and cellular response of UV light irradiated anodized titanium surface

  • Park, Kyou-Hwa;Koak, Jai-Young;Kim, Seong-Kyun;Heo, Seong-Joo
    • The Journal of Advanced Prosthodontics
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    • v.3 no.2
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    • pp.63-68
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    • 2011
  • PURPOSE. The object of this study was to investigate the effect of UV irradiation (by a general commercial UV sterilizer) on anodized titanium surface. Surface characteristics and cellular responses were compared between anodized titanium discs and UV irradiated anodized titanium discs. MATERIALS AND METHODS. Titanium discs were anodized and divided into the following groups: Group 1, anodized (control), and Goup 2, anodized and UV irradiated for 24 hours. The surface characteristics including contact angle, roughness, phase of oxide layer, and chemical elemental composition were inspected. The osteoblast-like human osteogenic sarcoma (HOS) cells were cultured on control and test group discs. Initial cellular attachment, MTS-based cell proliferation assay, and ALP synthesis level were compared between the two groups for the evaluation of cellular response. RESULTS. After UV irradiation, the contact angle decreased significantly (P<.001). The surface roughness and phase of oxide layer did not show definite changes, but carbon showed a considerable decrease after UV irradiation. Initial cell attachment was increased in test group (P=.004). Cells cultured on test group samples proliferated more actively (P=.009 at day 2, 5, and 7) and the ALP synthesis also increased in cells cultured on the test group (P=.016 at day 3, P=.009 at day 7 and 14). CONCLUSION. UV irradiation induced enhanced wettability, and increased initial cellular responses of HOS cells on anodized titanium surface.

Effects of Interleukin-1${\beta}$ and Tumor Necrosis $Factor-{\alpha}$ on the Release of Collagenase and Gelatinase from Osteoblasts

  • Eun, Jong-Gab;Baek, Dong-Heon;Kim, Se-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.5
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    • pp.269-274
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    • 2002
  • A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.