• 제목/요약/키워드: MC3T3-E1

검색결과 269건 처리시간 0.028초

Chitosan-PLLA 다층 다공성 차폐막의 생체적합성, 골세포활성도 및 골재생 능력에 관한 연구 (Evaluation on the biocompatibility, bone cell activity and bone regenerative capacity of chitosan-PLLA bilayer porous membrane)

  • 박준범;남성헌;김경화;이상철;이승진;김태일;설양조;이용무;구영;류인철;한수부;정종평
    • Journal of Periodontal and Implant Science
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    • 제35권3호
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    • pp.549-561
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    • 2005
  • 이 연구의 목적은 새로이 제작된 chitosan-poly(L-lactic acid)(PLLA) 다층 다공성 차폐막의 생체적합성 및 골세포활성도 및 골재생능을 평가하는 것이다. 제작된 차폐막을 24 well에 넣고 clonal osteoblast-like cell line(MC3T3-E1)을 접종한 군을 실험군으로, 차폐막을 사용하지 않은 대조군으로 하였다. 배양 1일, 7일 및 14일째에 각 well에서 세포수를 측정하였다. 주사전자현미경을 이용하여 차폐막에 부착된 세포의 형태관찰을 시행하였다. RNA 추출 및 RT-PCR을 실시한 후, agarose gel상에서 전기영동하여 조골세포 표식자인 collagen type I(COL), osteopontin(OP) 및 osteocalcin(OC) mRNA의 발현을 관찰하였다. 제작된 매트릭스의 생체적합성 및 골재생능을 관찰하기 위하여 백서의 두개골에 직경 8mm의 원형 결손부를 형성한 후 차폐막을 이식한 군을 실험군으로, 아무 것도 넣지 않은 군을 대조군으로 하여 4주 경과 후 실험동물을 희생시킨 후 조직학적관찰을 시행하였다. 시간경과에 따른 부착세포수 관찰결과, 배양 14일까지 조골세포의 수가 지속적으로 증가하였고, 주사전자현미경으로 세포의 형태 관찰결과, 배양된 세포들은 중층의 형태로 성장하면서 시간경과에 따라 세포가 응집되는 양상을 나타내었다. 관찰 기간동안 COL, OP, 및 OC mRNA의 발현이 관찰되어 배양 전 기간동안 조골세포의 형질이 잘 유지되고 있음을 알 수 있었다. 백서 두개골 결손부에 이식된 차폐막은 염증반응 없이 주위 조직과 우수한 생체적합성을 나타내었으며, 차폐막을 이식하지 하지 않은 대조군에 비해 높은 신생골 형성을 나타내었다. 이상의 관찰결과로 새로이 제작된 chitosan-PLLA 차폐막은 우수한 생체적합성 및 골재생능을 나타냄을 알 수 있었으며, 향후 이를 골조직 재생 및 치주조직유도재생 분야에 응용될 수 있을 것으로 생각된다.

$TNF{\alpha}$ Increases the Expression of ${\beta}2$ Adrenergic Receptors in Osteoblasts

  • Baek, Kyung-Hwa;Lee, Hye-Lim;Hwang, Hyo-Rin;Park, Hyun-Jung;Kwon, A-Rang;Qadir, Abdul S.;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • 제36권4호
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    • pp.173-178
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    • 2011
  • Tumor necrosis factor alpha ($TNF{\alpha}$) is a multifunctional cytokine that is elevated in inflammatory diseases such as atherosclerosis, diabetes and rheumatoid arthritis. Recent evidence has suggested that ${\beta}2$ adrenergic receptor (${\beta}2AR$) activation in osteoblasts suppresses osteogenic activity. In the present study, we explored whether $TNF{\alpha}$ modulates ${\beta}AR$ expression in osteoblastic cells and whether this regulation is associated with the inhibition of osteoblast differentiation by $TNF{\alpha}$. In the experiments, we used C2C12 cells, MC3T3-E1 cells and primary cultured mouse bone marrow stromal cells. Among the three subtypes of ${\beta}AR$, ${\beta}2$ and ${\beta}3AR$ were found in our analysis to be upregulated by $TNF{\alpha}$. Moreover, isoproterenol-induced cAMP production was observed to be significantly enhanced in $TNF{\alpha}$-primed C2C12 cells, indicating that $TNF{\alpha}$ enhances ${\beta}2AR$ signaling in osteoblasts. $TNF{\alpha}$ was further found in C2C12 cells to suppress bone morphogenetic protein 2-induced alkaline phosphatase (ALP) activity and the expression of osteogenic marker genes including Runx2, ALP and osteocalcin. Propranolol, a ${\beta}2AR$ antagonist, attenuated this $TNF{\alpha}$ suppression of osteogenic differentiation. $TNF{\alpha}$ increased the expression of receptor activator of NF-${\kappa}B$ ligand (RANKL), an essential osteoclastogenic factor, in C2C12 cells which was again blocked by propranolol. In summary, our data show that $TNF{\alpha}$ increases ${\beta}2AR$ expression in osteoblasts and that a blockade of ${\beta}2AR$ attenuates the suppression of osteogenic differentiation and stimulation of RANKL expression by $TNF{\alpha}$. These findings imply that a crosstalk between $TNF{\alpha}$ and ${\beta}2AR$ signaling pathways might occur in osteoblasts to modulate their function.

Inhibitory Effect of Biotransformed-Fucoidan on the Differentiation of Osteoclasts Induced by Receptor for Activation of Nuclear Factor-κB Ligand

  • Park, Bobae;Yu, Sun Nyoung;Kim, Sang-Hun;Lee, Junwon;Choi, Sung Jong;Chang, Jeong Hyun;Yang, Eun Ju;Kim, Kwang-Youn;Ahn, Soon-Cheol
    • Journal of Microbiology and Biotechnology
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    • 제32권8호
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    • pp.1017-1025
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    • 2022
  • Bone homeostasis is regulated by constant remodeling through osteogenesis by osteoblasts and osteolysis by osteoclasts and osteoporosis can be provoked when this balance is broken. Present pharmaceutical treatments for osteoporosis have harmful side effects and thus, our goal was to develop therapeutics from intrisincally safe natural products. Fucoidan is a polysaccharide extracted from many species of brown seaweed, with valuable pharmaceutical activities. To intensify the effect of fucoidan on bone homeostasis, we hydrolyzed fucoidan using AMG, Pectinex and Viscozyme. Of these, fucoidan biotransformed by Pectinex (Fu/Pec) powerfully inhibited the induction of tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts differentiated from bone marrow macrophages (BMMs) by the receptor for activation of nuclear factor-κB ligand (RANKL). To investigate potential of lower molecular weight fucoidan it was separated into >300 kDa, 50-300 kDa, and <50 kDa Fu/Pec fractions by ultrafiltration system. The effects of these fractions on TRAP and alkaline phosphatase (ALP) activities were then examined in differentiated osteoclasts and MC3T3-E1 osteoblasts, respectively. Interestingly, 50-300 kDa Fu/Pec suppressed RANKL-induced osteoclasts differentiation from BMMs but did not synergistically enhance osteoblasts differentiation induced by osteogenic agents. In addition, this fraction inhibited the expressions of NFATc1, TRAP, OSCAR, and RANK, which are all key transcriptional factors involved in osteoclast differentiation, and those of Src, c-Fos and Mitf, as determined by RT-PCR. In conclusion, enzymatically low-molecularized 50-300 kDa Fu/Pec suppressed TRAP by downregulating RANKL-related signaling, contributing to the inhibition of osteoclasts differentiation, and represented a potential means of inducing bone remodeling in the background of osteoporosis.

In vitro 검출 시스템을 이용한 한약재 추출물들의 에스트로겐 유사활성 검증 (Verification of Estrogen Like Activities of Herbal Medicines Using an In Vitro Detection System)

  • 박성환;전명제;장민경;이솔지;김보경;전명정;김서연;김미향;이동근;이태호;남재섭;허지원;이상현
    • 동의생리병리학회지
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    • 제27권6호
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    • pp.752-758
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    • 2013
  • Estrogen like activities were evaluated using ethanol and hot water extracts of herbal medicines by using an in vitro detection system. Bokryung (Poria cocos), Sanyak (root of Dioscorea batatas) and Mokdanpi (root skin of Paeonia suffruticosa) represented statistically significant estrogen-like activities (p<0.001), while Omija (fruit of Schizandra chinensis), Taeksa (root of Alisma canaliculatum A. BR.), Jihwang (root of Rhemannia glutinosa), and Sansuyu (fruit of Cornus officinalis) did not. Estrogen-like activities of Bokryung hot water extract (500 ${\mu}g/ml$) and ethanol extract (50 ${\mu}g/ml$) were almost same as that of a C M $17{\beta}$-estradiol. Furthermore, estrogen-like activities of ethanol extracts (500 ${\mu}g/ml$) of Bokryung and Mokdanpi were stronger than that of $10^{-7}$ M $17{\beta}$-estradiol. These results suggest that Bokryung, Sanyak and Mokdanpi show estrogen-like activities. Especially, Sanyak and Mokdanpi represented promotive effect on the proliferation of MC3T3-E1 osteoblastic cells. Bokryung, Sanyak and Mokdanpi also exhibited superior inhibitory effect on the viability of RAW 264.7 cells. In conclusion, these three herbal medicines might be interpreted as candidates for the further study or development of functional foods or medicine to prevent or avoid postmenopausal symptoms of women.

임플랜트에 연결한 영구자석이 임플랜트 주위 뼈모세포의 분포와 부착에 미치는 영향에 관한 연구 (THE EFFECT OF PERMANENT MAGNET CONNECTING WITH DENTAL IMPLANT ON DISTRIBUTION AND ATTACHMENT OF OSTEOBLAST-LIKE CELL AROUND THE DENTAL IMPLANT)

  • 오나희;최부병;권긍록;백진;이성복
    • 대한치과보철학회지
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    • 제43권4호
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    • pp.511-518
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    • 2005
  • Purpose: The purpose of this study is to find the effect of rare earth magnet's magnetic field of to the osteoblast around the implant by the means of observation number, and distribution around the implant which is connected to the permanent magnet but not, counted and compared by the number of cells attached to the surface of the implant. Material and method: The permanent magnets, made in the healing cap form, were connected to the implant future, and placed on the culture plate, The osteoblast-like cell: MC3T3-E1 were used for cell culture. As the control group, the implant were connected to normal healing cap, and cultured in the same conditions. 48 hours later, using inverted microscope, the number and distribution of osteoblast around the implant were observed, and 72 hours later, the number of the cells attached to the implant were counted. Results: As a result, the implant connected to the permanent magnet had proved to have a more concentrated cell distribution rate than the control group. The implant connected to the permanent magnet, neck area : which has about 10 gauss magnetic force, had more cells than apex area. The implant connected to the permanent magnet had proven to attach to the osteoblast more productively than control group's implant. Conclusions: This research showed that the magnetic field of the permanent magnet affected the distribution and growth rate of the osteoblast around the implant. In order to support this study, it also had need to monitor the progress of the permanent magnet specifically shown on the neck area, which has10 gauss magnetic force. So after additional research on the distribution and attachment of the cells, and further more, on bone formation, it will be concluded that the clinical applications ,such as immediate loading of implant treatment are possible.

난소 적출 흰쥐 골다공증 모델에서 금은화(金銀花)가 골밀도 증가에 미치는 효과 (Effect of Lonicerae Japonicae Flos on Bone Density in Ovariectomized Rat Model of Osteoporosis)

  • 이성엽;김민선;홍수연;김재현;김홍식;이충호;정혁상;손영주
    • 대한본초학회지
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    • 제36권5호
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    • pp.81-91
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    • 2021
  • Objectives : Osteoporosis is a systemic skeletal disease that decreases bone density and increases the risk of fractures. Bisphosphonates and SERMs are mainly used to treat osteoporosis, but, long-term use increases the risk of side effects such as jaw bone necrosis and breast cancer. Therefore, it is necessary to develop a therapeutic agent for a natural product with few side effects. Water extract of Lonicerae Japonicae Flos (wLF) was mainly found to have anti-cancer and anti-inflammatory effects. However, the effect of wLF on osteoporosis has not been elucidated. Therefore, this experiment investigated the effect of wLF on osteoclasts, osteoblasts and osteoporosis models. Methods : In order to study the effect of wLF on osteoporosis, the OVX-induced rat model was used for in vivo study. After 8 weeks, we measured body weight, uterine weight, liver weight, femur weight, bone density, trabecular area and tibia ash weight. To determine the effect of wLF on osteoclast differentiation, we measured the number of TRAP-positive cells and TRAP activity. To examine the effect of wLF on the expression of osteoblast-related genes, we measured the mRNA expression of alkaline phosphatase (ALP, Alpl) and osteocalcin (OCN, Bglap2). Results : In vivo experiment, wLF inhibited the reduction of femur weight, trabecular area, bone density and tibia ash weight. In vitro experiment, wLF had no significant effect on osteoclast differentiation. However, wLF increased the mRNA expression of Alpl and Bglap2 in MC3T3-E1 cell. Conclusions : This result suggested that wLF may be used for the treatment and prevention of postmenopausal osteoporosis.

오적산의 추출 용매 차이에 따른 염증 및 염증 관련 질환에 대한 효능 비교 (Effects of water and ethanol extracts from Ojeok-san on inflammation and its related diseases)

  • 유새롬;정수진;김예지;임혜선;진성은;전우영;신인식;신나라;김성실;김정훈;하혜경;이미영;김온순;서창섭;신현규
    • 대한한방내과학회지
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    • 제33권4호
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    • pp.418-428
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    • 2012
  • Objectives : Ojeok-san, a traditional herbal formula, has been used for the treatment of cold illness and its related symptoms such as headache, nausea and indigestion. This study was performed to compare effects of water (OJSW) and 70% ethanol extracts (OJSE) of Ojeok-san on inflammation and its related diseases atopy, asthma and obesity in vitro. Methods : We performed HPLC to investigate contents of index components of OJSW and OJSE. We investigated the effects of OJSW and OJSE with an in vitro model, using 5 cell lines, specifically RAW 264. 7, HaCaT, MC/9, BEAS-2B and 3T3-L1. Results : HPLC analysis displayed that the contents of index components were higher in OJSE than OJSW. In lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, OJSE significantly inhibited productions of interleukin (IL)-6, nitrite and prostaglandin $E_2$ ($PEG_2$). In TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT keratinocytes, OJSE significantly lowered levels of macrophage-derived chemokine (MDC) as well as regulated and normal T cell expressed and secreted (RANTES). OJSE also had a protective effect on inflammatory response by decreasing RANTES secretion in TNF-${\alpha}$-stimulated BEAS-2B cells. Conclusions : We conclude that OJSE could be more appropriate to enhance the biological activities against inflammation and its related diseases, and could be applied as a bioactive material for developing the potent anti-inflammatory agents.

희토류 자석의 자성이 골모세포 성장인자 수용체의 증가에 미치는 영향에 관한 연구 (Effect of the magnetism(neodymium magnet) on growth factor receptors of osteoblasts)

  • 이상민;이성복;최부병
    • 구강회복응용과학지
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    • 제19권2호
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    • pp.87-96
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    • 2003
  • The purposes of this study were to find out the optimum intensity of magnetic field where magnetism could promote the activity of osteoblast, and to discover the possibility of clinical application in the areas of dental implants and bone grafts by confirming the effect of clinically increasing bone formation. In this experiment, we used the Neodymium magnet, which had magnetic power six times as strong as the current ones and enabled the resistances against the demagnetization up to 20 to 50 times to be minimized with the size of 1mm in sight. In order to culture cells, a specially designed device was used. It was made to adjust the distance and accordingly to control the intensity of the magnetic field, by placing the cell culture plate in the center with a magnet of 1mm long and thick installed on the both ends. Using MC3T3-E1 cell, a kind of osteoblast-like cell, we cultured, for 24 hours, not only the test group which had been cultured under the magnetic fields with different intensity of 5, 10, 50, 100, 500, and 1000 Gauss, but also the control group excluding the influences of the magnetic field. After observing the cell's form and the density of the culture medium through an inverted microscope, we made a series of proceedings needed for the immunofluoroscence staining, such as fixation, normal serum reaction, primary antibody reaction, and secondary antibody reaction. And with a fluorescence microscope, we observed those-above and compared the frequency of expression of IFG-1 receptor. To make a Western immunoblotting analysis, the cells cultured under the same condition as the above had the procedure of the lysis buffer and the acrylamide gel electrophoresis was carried out. Protein transferred into the nitrocellulose membrane and tested on the primary and the secondary antibody reactions was observed and compared. The results were as follows: When observed through an inverted microscope, the nuclear divisions of the cells under the magnetic field of 10 Gauss were the most active, and the density of the cells could be observed the most enormously. As the result of an immunofluoroscence staining of IGF-1 receptor, the expression of IFG-1 was the most frequently observed under the magnetic field of 10 Gauss. On the other hand, few differences of consideration were made between the test group cultured under the magnetic fields of 5, 500, and 1000 Gauss and the control group. In respect of the expression of IFG-1 receptor, the test group cultured under the magnetic fields of 50 and 100 Gauss were higher than the control group, and lower than that cultured under the magnetic field of 10 Gauss.(p<0.05) According to the Western immunoblotting analysis, the band of IFG-1 receptor which had 85KDa of molecular weight was the darkest. Judging from the above-mentioned results, the growth factor receptor of an osteoblast cell which was an important criterion for the bone formation was increased in maximum under the magnetic field of 10 Gauss. Moreover it was observed that the optimum intensity of magnetic field in which magnetism made the activity of the osteoblast cell increase was about 10 Gauss.

합성곱 신경망의 비지니스 응용: 런웨이 이미지를 사용한 의류 분류를 중심으로 (Business Application of Convolutional Neural Networks for Apparel Classification Using Runway Image)

  • 서이안;신경식
    • 지능정보연구
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    • 제24권3호
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    • pp.1-19
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    • 2018
  • 최근 딥러닝은 오디오, 텍스트 및 이미지 데이터와 같은 비 체계적인 데이터를 대상으로 다양한 추정, 분류 및 예측 문제에 사용 및 적용되고 있다. 특히, 의류산업에 적용될 경우 딥러닝 기법을 활용한 의류 인식, 의류 검색, 자동 제품 추천 등의 심층 학습을 기반으로 한 응용이 가능하다. 이 때의 핵심모형은 합성곱 신경망을 사용한 이미지 분류이다. 합성곱 신경망은 입력이 전달되고 출력에 도달하는 과정에서 가중치와 같은 매개 변수를 학습하는 뉴런으로 구성되고, 영상 분류에 가장 적합한 방법론으로 사용된다. 기존의 의류 이미지 분류 작업에서 대부분의 분류 모형은 의류 이미지 자체 또는 전문모델 착용 의류와 같이 통제된 상황에서 촬영되는 온라인 제품 이미지를 사용하여 학습을 수행한다. 하지만 본 연구에서는 통제되지 않은 상황에서 촬영되고 사람들의 움직임과 다양한 포즈가 포함된 스트릿 패션 이미지 또는 런웨이 이미지를 분류하려는 상황을 고려하여 분류 모형을 훈련시키는 효과적인 방법을 제안한다. 이동성을 포착하는 런웨이 의류 이미지로 모형을 학습시킴으로써 분류 모형의 다양한 쿼리 이미지에 대한 적응력을 높일 수 있다. 모형 학습 시 먼저 ImageNet 데이터셋을 사용하여 pre-training 과정을 거치고 본 연구를 위해 수집된 32 개 주요 패션 브랜드의 2426개 런웨이 이미지로 구성된 데이터셋을 사용하여 fine-tuning을 수행한다. 학습 과정의 일반화를 고려해 10번의 실험을 수행하고 제안된 모형은 최종 테스트에서 67.2 %의 정확도를 기록했다. 본 연구 모형은 쿼리 이미지가 런웨이 이미지, 제품 이미지 또는 스트릿 패션 이미지가 될 수 있는 다양한 분류 환경에 적용될 수 있다. 구체적으로는 패션 위크에서 모바일 어플리케이션 서비스를 통해 브랜드 검색을 용이하게 하는 서비스를 제공하거나, 패션 잡지사의 편집 작업에 사용되어 브랜드나 스타일을 분류하고 라벨을 붙일 수 있으며, 온라인 쇼핑몰에서 아이템 정보를 제공하거나 유사한 아이템을 추천하는 등의 다양한 목적에 적용될 수 있다.