• Title/Summary/Keyword: 세포부착

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The biological effects of fibronectin typeIII 7-10 to MC3T3-E1 osteoblast (Fibronectin type III 7-10 이 조골세포에 미치는 영향)

  • Hong, Jeong-Ug;Choi, Sang-Mook;Han, Soo-Boo;Chung, Chong-Pyoung;Rhyu, In-Chul;Lee, Yong-Moo;Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.143-160
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    • 2002
  • 타이태늄은 뛰어난 생체적합성과 적절한 물리적 성질을 바탕으로 치과 및 정형외과 영역의 매식재로 널리사용되어져 왔으며, 골과 매식재 사이의 골 융합 정도를 증가시킬 목적으로 물리, 화학적인 방법을 이용한 타이태늄의 표면처리에 관한 많은 연구들이 진행되어 왔다. 최근에는 부착단백질 또는 성장인자를 이용한 생체재료의 표면개질을 통하여 조직적합성 및 치유 능의 개선을 위한 시도들이 있어왔다. Fibronectin(FN)은 주요 세포외기질중의 하나로 생체 내 널리 분포하여 세포의 부착, 이동 및 증식에 관여하는 거대 당단백으로, RGD및 PHSRN 펩타이드 서열이 세포의 인테그린과 결합하여 세포의 활성을 조절하는 것으로 알려져 있다. 이 연구에서는 FN으로 처리된 타이태늄이 조골세포의 부착, 증식 및 분화에 미치는 영향과 이에 따른 석회화 정도에 미치는 영향을 관찰하여 부착분자를 이용한 타이태늄 표면개질의 효과를 규명하고자 하였다. 상업용 순수 타이태늄을 gold thiol법을 이용하여 표면처리 후, 혈장 FN(plasma FN, pFN)과 유전자재조합법을 이용하여 얻은 FN조각(FN type III 7-10, FNIII 7-10)을 피복한 시편을 실험군으로, 아무런 처리를 하지 않은것(smooth surface, SS)과 산 부식(Sandblasted and acid etched, SLA)처리된것을 대조군으로 이용하였다. 배양된 조골세포주(MC3T3-E1)를 사용하여 타이태늄 표면 처리에 따른 세포의 증식, 형태변화, 알칼리성 인산분해효소(ALPase) 생산 및 세포면역형광법을 이용한 분화정도를 시간 경과에 따라 관찰하였다. 조골세포증식의 경우 FNIII 7-10 처리군에서 pFN 처리군 및 대조군에 비해 시간경과에 따라 유의성있는 세포수의 증식이 관찰되었으며(p<0.05), ALPase 생성의 경우에도 FNIII 7-10 처리 군에서 아무 처리도 하지 않은 군에 비해 유의성 있게 높은 효소의 생성이 관찰되었다(p<0.05). 주사전자현미경을 이용한 세포의 형태관찰결과 아무 처리도 하지 않은 군에서는 마름모형태를 나타내었으며, 산 부식 처리된 군에서는 세포가 가시모양의 형태를 보인 반면 FN으로 처리된 두 군에서는 세포의 부착 및 펴짐이 매우 발달되어 있는 모습이 관찰되었다. 세포의 분화정도를 관찰하기 위하여 국소부착키나제(focal adhesion kinase, FAK), 및 actin stress fiber의 분포양상을 세포면역형광법을 이용하여 관찰한 결과 FN으로 표면처리된 두 군에서 아무런 처리도 하지않은 군 및 산 부식처리 한 군에 비해 프라크의 발현이 높게 나타났으며 잘 발달된 actin stress fiber의 소견을 나타내었다. 이 실험의 결과들은 gold thiol 법을 이용한 표면처리 후 FN부착을 통한 타이태늄의 표면개질이 조골세포의 부착, 증식 및 분화에 중요한 역할을 담당하여 석회화 정도를 촉진시키는 것을 보여주었으며, 이런 결과들은 더 짧은 FN조각을 이용한 다른 생체재료의 표면개질에 폭 넓게 응용할 수 있으리라 생각된다.

Anti-Cell Adhesion Effect of Animal Cell with Regional Special Natural Products of Anthrisci radix, Psoraleae semen, Siegesbeckiae herba and Corni fructus (지역 특산 천연산물 전호, 파고지, 희첨 및 산수유의 동물세포간의 부착 억제효과)

  • Shin, Jin-Hyuk;Cha, Gu-Yong;Kim, Hui-Jin;Hwang, Jae-Ho;Han, Kyeong-Ho;Seo, Hyo-Jin;Kim, Min-Yong;Oh, Suk-Jung;Kim, Jong-Deog
    • KSBB Journal
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    • v.24 no.6
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    • pp.541-548
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    • 2009
  • For investigation of anti-angiogenesis mechanism of Anthrisci radix, Psoraleae semen, Siegesbeckiae herba and Corni fructus, anti-cell adhesion experiment was performed. The adhesion of U937 cells to IL-$1\beta$-stimulated HUVECs was completely suppressed by 276% at 0.2 mg/L of Anthrisci radix, 220% at 5 mg/L of Psoraleae semen, 158% at 10 mg/L of Siegesbeckiae herba and 132% at 20 mg/L of Corni fructus, respectively. And the adhesion of PMA-chemical stimulated U937 cells to HUVECs, it was inhibited 139% at 0.2 mg/L of Anthrisci radix, 442% at 5 mg/L of Psoraleae semen, 720% at 10 mg/L of Siegesbeckiae herba and 664% at 20 mg/L of Corni fructus. Also, the adhesion of chemical stimulated U937 cells to IL-$1\beta$/chemical stimullated HUVECs, it was inhibited by 286% at 0.2 mg/L of Anthrisci radix, 146% at 5 mg/L of Psoraleae semen, 436% at 10 mg/L of Siegesbeckiae herba and 297% at 20 mg/L of Corni fructus, respectively. It would be a useful substance for anti-cell adhesion based on anti-angiogenesis for anti-obesity and anti-cancer.

Beta-4 Integrin Transfection, Cloning and Functional Assay in Squamous Cell Carcinoma (Beta-4 Integrin 유전자 주입, 클로닝과 편평상피암에서의 Beta-4 Integrin 기능에 관한 연구)

  • Kim Young-Min;Carey Tomas E.
    • Korean Journal of Head & Neck Oncology
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    • v.13 no.2
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    • pp.169-179
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    • 1997
  • 서론 : Laminin의 수용기로 알려진 Integrin $\alpha6\beta4$의 세포내 표현 정도는 편평상피암을 위시한 여러 악성종양의 전이능력 및 예후와 밀접한 상관관계가 없다고 알려져 있다. 이 Integrin은 Laminin과 같은 세포와 리간드와 결합하면 상피세포의 기저막 지주 구조물인 hemidesmosome의 세포체질 요소(cytoskeletal element)와 연관되어 그 결과 세포의 기저막과 세포내 케라틴을 연결하는 역할을 한다. Integrin $\alpha6\beta4$는 구조적으로 다른 많은 integrin들과 달리 $\beta$4의 세포질내 영역(cytoplasmic domain)이 특징적으로 크다. 이 세포질내 영역 $\beta$4 integrin의 기능은 아직 밝혀지지 않고 있으나 아마 세포 성장의 신호전달 및 악성종양의 특징인 침윤 전이에 관련할 것으로 보아지고 있다. 재료 및 방법: 저자들은 우선 $\beta$4 integrin의 wild type s-DNA와 $\beta$4 세포질내 영역(cytoplasmic domain) 및 $\beta$4의 tyrosine 인산화 반응 부위가 각각 결손된 c-DNA를 PCR을 통하여 합성하여 pRc/CMV 벡터에 삽입한 후 원래 $\beta$4 integrin의 발현이 결집된 인간 방광암 세포에 Calcium phosphate precipitation 방법으로 주입(transfection)시켜 형질변환된 세포를 면역형광법, Flow cytometry 및 Immunoprecipitation 방법으로 클로닝하여 wild type $\beta$4-full length(Clone FL), truncated $\beta$4-cytoplasmic domain(C1one CD), 및 mutated $\beta$4-tyrosine phosphorylation site (Clone M)을 얻었다. 암 세포의 부착 및 침투 능력의 기능적 연구로 모노 클로날 항체와 fibronectin, laminin, Matrigel을 단백질 기질로 사용하였으며 결과 비교를 위하여 pRc/CMV 벡터만 주입시켰던 클로운과 방광암 세포주를 $\beta$4 integrin 음성 대조군으로 또한 이 Integrin의 높은 발현을 보이는 두경부 편평상피암 세포주를 양성 대조군으로 이용하였다. 결과 : 세포부착능력에 있어서 온전한 $\beta$4 cytoplasmic domain이 존재하는 클로운이 laminin에 강한 부착능력을 보였으나 fibronectin의 부착정도는 $\beta$4 integrin의 표현정도와 관계없이 모든 클로운에서 비슷하였다. Matrigel을 투과하는 암세포 침윤 능력에서는 $\beta$4 integrin의 표현이 존재하는 클로운들이 투과 능력이 높았으나 세포외 리간드가 없는 control membrane을 사용하였을 때와 비교하여 투과능력의 차이를 보이지 않았다. 결론 : 유전자 주입(transfection) 방법으로 integrin의 다양한 클로운의 합성이 가능하여 이 Integrin의 암 세포의 부착 및 침투 능력에서의 기능을 규명 할 수 있게 한다. $\beta$4 integrin은 편평상피 암세포의 부착에 있어서 세포외 리간드 laminin과 특이 결합하여 부착 능력을 높이는 중요한 역할을 하며 편평상피 암세포의 침투에 있어서는 $\beta$4 integrin의 표현이 침투 능력을 높이는 역할을 하나 이때에는 laminin과 같은 리간드와의 특이 결합에 의존하지는 않는 것으로 사료된다.

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Improvement of an hydroxyapatite bead adherence assay for streptococcus sanguis (Streptococcus sanguis의 구형 Hydroxyapatite 비드에의 부착 Assay 방법의 개량)

  • 최선진;이시영;송요한
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.48-55
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    • 1989
  • The purpose of the present investigation was to improve several procedures being used in the adherence assay of Streptococcus sanguis cells to hydroxyapatite (HA) beads and to study the effect of the beads on the counting of radioactivity. The standard adhere assay involved the adherence of radiolabeled bacteria to 40mg of HA beads. The beads were mixed with ['H]thymidine-labeled bacterial cells and incubated for 60 minutes at room temperature. Unadsorbed cells were removed, the beads with adsorbed cells were dried, and the radioactivity was monitered in a scintillation spectrometer. The 30 seconds sonication of cells in a form of long chains appeared to be adequate for obtaining mostly singlet or doublet cells. Unlike the counting of S. sanguis cell suspension, bacterial cells adhered to HA or saliva-coated HA(SHA) required smaller volume (2.5ml) of scintillaton fluid for better counting. Eighteen percent quenching of counts could be attributed to the beads. Among 3 procedures commonly used to equilibrate the beads for adherence assay, no differences were found in their effectiveness. The HA beads on which the bacteria remained attached in scintillant during the counting were found to be the source of sample self-absorption representing 34.5% of the total radioactivity counts resulting from the beads dissolved in HCl solution.

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Adherence-induced gene expression in human alveolar macrophages (표면부착에 의한 사람 폐포대식세포의 유전자 발현에 관한 연구)

  • Chung, Man Pyo;Yoo, Chul Gyu;Han, Sung Koo;Shim, Young-Soo;Rhee, Chong H.;Han, Yang Chol;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.936-944
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    • 1996
  • Background: Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. Adhesion molecules and gene transcription of the inflammatory mediators are known to be associated in this process. To evaluate whether adhesion molecule and transcriptional activation of the inflammatory substances are also involved in the activation of human alveolar macrophage by the adherence procedure, we designed this experiment. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be nonnal by chest cr and alveolar macrophage was harvested. To measure the expression of Interleukin-8(IL-8) mRNA, manganese superoxide dismutase(SOD) mRNA and CD11/CD18 mRNA in human alveolar macrophage of both adherence state and suspension state, Northern blot analysis was done at 0, 2, 4, 8 and 24hrs after the adherence to plastic surface and during suspension state. Then, phorbol myristate acetate(pMA) and N-formyl-methionyl-leucyl-phenylalanine(fMLP) were added respectively in the same experimental condition. Result : 1) Human alveolar macrophages in the adherent state induced IL-8 mRNA and SOD mRNA expression which was maximal at 8 hours after the adherence to plastic surface. But we could not observe the upregulation of CD18 mRNA by surface adherence. 2) PMA induced these mRNA expression both in the adherent cell and the nonadherem cells, but the induction of mRNA expression by fMLP occurred only in the adherent cells. Conclusion: These results suggest that adherence of huamn alveolar macropahge is an important cell-activating event that may play a critical role in the modulation of lung inflammatory respones.

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Adhesion of Bifidobacteria to Caco-2 Cells and in Relation to Cell Surface Hydrophobicity (비피도박테리아의 Caco-2 세포에 대한 부착성과 세포 표면 소수성)

  • Lim, Kwang-Sei;Huh, Chul-Sung
    • Food Science of Animal Resources
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    • v.26 no.4
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    • pp.497-502
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    • 2006
  • The adhesion of 16 bifidobacterial strains, including 10 isolates from Korea infants, to Caco-2 cells and their cell surface hydrophobicity were tested. The results of adhesion and cell surface hydrophobicity of for various bifidobacterial strains were obtained and correlations between adhesion and hydrophobicity were strain-dependent properties. Any correlations between species of tested strains were not observed. Among the tested strains, Bifidobacterim longum D6, B. longum H4, B. thermophilum ATCC 25525, B. suis ATCC 27533, and B. animalis subsp. lactis BB12 had higher adherent properties and B. bifidum B3, B. longum D6, B. longum stronger hydrophobicity, respectively. Due to the strain-dependant correlation between adhesion to Caco-2 cells and cell surface hydrophobicity of bifidobacteria, these results provide a possible method for preliminary selection of bifidobacteria potentially adherent to Caco-2 cells by means of cell surface hydrophobic properties.

3,3'-Diindolylmethane (DIM) decrease adhesion, migration and invasion in human prostate cancer cells (3,3'-Diindolylmethane (DIM)이 인체 전립선암 세포의 부착, 이동 및 침윤성에 미치는 영향)

  • Kim, Hyeon-A
    • Food Science and Preservation
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    • v.22 no.1
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    • pp.19-26
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    • 2015
  • Indole 3-carbinol (I3C), important component of cruciferous vegetables and its major acid-catalyzed metabolite, 3,3'-diindolylmethane (DIM) have been suggested to have an inhibitory effect on the tumor growth and metastasis. This study investigated the effect of DIM on the adhesion, migration and invasion of highly invasive PC3 and DU145 human prostate cancer cell lines. Cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 3.0 g/L glucose, 3.7 g/L sodium bicarbonate and 10% fetal bovine and were incubated in a humidified incubator at $37^{\circ}C$ and 5% $CO_2$. DIM reduced the adhesion of PC3 and DU145 cells in a dose dependent manner. The pretreatment of PC3 cells with DIM reduced the adhesion dose dependantly, but inhibition was less effective than the treatment with DIM during the adhesion assay. The migration and invasion of PC3 and DU145 cells were reduced by DIM dose dependantly, and the inhibition of DIM was less effective in the DU145 cells than in the PC3 cells. The pretreatment of PC3 cells with DIM for 24 hr before the assay reduced invasion of PC3 cells by 37%. These results suggest that DIM inhibits adhesion, migration and invasion of the PC3 and DU145 cells and may be an effective antimetastatic therapy in addition to traditional chemotherapy.

Cell Patterning on Various Substrates Using Polyelectrolyte Multilayer and Microstructure of Poly(Ethylene Glycol) (다양한 기판 위에서 고분자 전해질 다층 막과 폴리에틸렌글리콜 미세 구조물을 이용한 세포 패터닝 방법)

  • Shim, Hyun-Woo;Lee, Ji-Hye;Choi, Ho-Suk;Lee, Chang-Soo
    • Korean Chemical Engineering Research
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    • v.46 no.6
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    • pp.1100-1106
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    • 2008
  • In this study, we presented rapid and simple fabrication method of functionalized surface on various substrates as a universal platform for the selective immobilization of cells. The functionalized surface was achieved by using deposition of polyelectrolyte such as poly(allyamine hydrochloride) (PAH), poly(diallyldimethyl ammonium chloride) (PDAC), poly(4-ammonium styrene sulfonic acid) (PSS), poly(acrylic acid) (PAA) and fabrication of poly(ethylene glycol) (PEG) microstructure through micro-molding in capillaries (MIMIC) technique on each glass, poly(methyl methacrylate) (PMMA), polystyrene (PS) and poly(dimethyl siloxane) (PDMS) substrate. The polyelectrolyte multilayer provides adhesion force via strong electrostatic attraction between cell and surface. On the other hand, PEG microstructures also lead to prevent non-specific binding of cells because of physical and biological barrier. The characteristic of each modified surface was examined by using static contact angle measurement. The modified surface onto several substrates provides appropriate environment for cellular adhesion, which is essential technology for cell patterning with high yield and viability in the micropatterning technology. The proposed method is reproducible, convenient and rapid. In addition, the fabrication process is environmentally friendly process due to the no use of harsh solvent. It can be applied to the fabrication of biological sensor, biomolecules patterning, microelectronics devices, screening system, and study of cell-surface interaction.

Study on attachment of bacteria to tonsillar epithelial cell during acute tonsillitis (급성편도선염에서 편도상피세포의 세균부착성에 관한 연구)

  • 이흥만;정형목;최충식;이우섭;이상학;황순재
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.98-98
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    • 1993
  • To cause invasive disease, microorganism must attach firmly to the tonsillar epithelial cell. Once attached, the microorganisms can proliferated, form colonies and release extracillular toxins which can injure the underlying cells. The purpose of present study was to asertain whether or not there exist in vivo differences in bacterial attachment between patients with acute tonsillitis and healthy individuals as a control. This study was carried out on 20 patients suffering from acute tonsillitis and 20 healthy persons used as control. After scraping of the surface of tonsil, cellular mixture was stained with Acridine orange and the number of attached bacteria was calculated using a fluorescent microscope. The adherence rate was calculated as number of bacteria attached to each of 50 epithelial cells. simultaneously, we peformed conventional bacterial culture. Conclusively, the attachment of more than 10% bacteria to the tonsillar epithelial cell was significantly greater in acute tonsillitis group than control group, and there was a significant correlation between age and the number of the attached bacteria to the epithlium.

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Attaching Nature and Community Variation of Epiphytic Diatoms on Leaf of Zostera spp. (Zostera속 해초에 부착하는 규조류의 부착 특성과 해초 종별 군집 변화)

  • Chung, Mi-Hee;Youn, Seok-Hyun;Yoon, Won-Duk
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.4
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    • pp.184-191
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    • 2010
  • Epiphytic diatoms are very important organisms in the seagrass ecosystem because their colonization on leaves increases microtopography and provides attachment sites that make the leaves more hospitable for other epiphytes. Epiphytic diatoms were attached to the leaves in the following 3 manners: (1) parallel to the cells of the seagrass leaf or by molding the shape of the diatom along the cell shape of the leaf; (2) with increasing diatom density toward the leaf tip; (3) Cocconeis species as attaching species than the Naviculoid species as the second attaching species on the leaf tip. In addition, the epiphytic diatom communities on Zostera marina leaves differed from those on the Zostera japonica leaves, but were very similar to the epiphytic communities on Zostera caespitosa leaves. Our results suggest that the epiphytic community on seagrass leaves varied according to the leaf shape such as leaf length and width, but the leaf cell shape or size did not influence the dynamics of the diatom communities.