• Title/Summary/Keyword: Adhesion assay

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Characterization of Adhesion of Bifidobacterium sp. BGN4 to Human Enterocyte-Like Caco-2 Cells

  • Kim, In-Hee;Park, Myung-Soo;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.276-281
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    • 2003
  • The adhesion of probiotic bacteria to the intestinal mucosa is one of the desirable properties for their colonization in the intestinal tract, where these bacteria constantly compete with other bacteria. The adhesion of different strains of bifidobacteria to Caco-2 cells was compared. Among the strains examined, BGN-4 showed the highest adhesion level and the greatest cell surface hydrophobicity (CSH). No close relationship was found between the adhesion and CSH of the strains. Upon protease and heat treatment, the adhesion of the BGN-4 to the Caco-2 cells decreased significantly. The cells grown at $42^{\circ}C$ showed a lower CSH and self-aggregation levels than cells grown at $37^{\circ}C$. The treatment of EGTA did not have any effect on the adhesion. The degree of adhesion did not differ among the experimental groups in which galactose, mannose, or fucose were added in the adhesion assay mixture. The results suggest that the adhesion of the Bifidobacterium to the epithelial cells may be affected by the composition and structure of the cell membrane and interacting surfaces.

Biofilm-forming ability and adherence to poly-(methyl-methacrylate) acrylic resin materials of oral Candida albicans strains isolated from HIV positive subjects

  • Uzunoglu, Emel;Bicer, Arzu Zeynep Yildirim;Dolapci, Istar;Dogan, Arife
    • The Journal of Advanced Prosthodontics
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    • v.6 no.1
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    • pp.30-34
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    • 2014
  • PURPOSE. This study evaluated the adhesion to acrylic resin specimens and biofilm formation capability of Candida albicans strains isolated from HIV positive subjects' oral rinse solutions. MATERIALS AND METHODS. The material tested was a heat-cured acrylic resin (Acron Duo). Using the adhesion and crystal violet assays, 14 oral Candida albicans isolated from HIV-positive subjects and 2 references Candida strains (C. albicans ATCC 90028 and C. albicans ATCC 90128) were compared for their biofilm production and adhesion properties to acrylic surfaces in vitro. RESULTS. There were no significant differences in adhesion (P=.52) and biofilm formation assays (P=.42) by statistical analysis with Mann-Whitney test. CONCLUSION. Denture stomatitis and increased prevalence of candidal carriage in HIV infected patients is unlikely to be related to the biofilm formation and adhesion abilities of C. albicans to acrylic resin materials.

Cell-cell Adhesion of Jurkat T Cells Induced by CD29 and CD98 Activation and its Application (CD29 및 CD98 활성 매개에 의한 Jurkat T 세포의 유착과 그 활용)

  • Kim, Byung-Hun;Cho, Jae-Youl
    • YAKHAK HOEJI
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    • v.53 no.3
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    • pp.119-124
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    • 2009
  • Cell-cell adhesion managed by various adhesion molecules plays an important role in regulating functional activation of cells. This event mediates attachment of inflammatory cells to endothelial cells, interaction of antigen-presenting cells with T cells and metastatic adherence of cancer cells to epithelial tissue cells. Therefore, this cellular response is considered as one of therapeutic target to treat various cancers and inflammatory diseases. To develop proper model for evaluation of functional activation of adhesion molecules, the ability of U937 and Jurkat T cells responsive to various adhesion inducers such as phorbal-12-myristate-13-acetate (PMA), staurosporin and monoclonal antibodies to CD29, CD43 and CD98 was investigated using quantitative cell-cell adhesion assay. U937 cells made more cell-cell clusters by the treatment of antibodies to CD29 and CD43 than Jurkat T cells, while Jurkat T cells exhibited increased cell-cell adhesion ability in CD98 antibody treatment. In agreement, the surface levels of CD29 and CD98 were highly observed in U937 and Jurkat T cells, respectively. Therefore, our data suggest that Jurkat T and U937 cells can be used for model system to evaluate functional activation of adhesion molecules such as CD29 and CD98.

A Study on the Cytotoxic Effects of Several Plant Extracts on the Cell viability and Cell Adhesion Activity in Cultured NIH3T3 Fibroblast (몇 가지 식물추출물이 배양 NIH3T3 섬유모세포의 세포생존율과 세포부착률에 미치는 세포독성에 관한 연구)

  • Rim, Yo-Sup;Song, Won-Seob;Seo, Young-Mi;Park, Seung-Taeck;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.3
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    • pp.116-124
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    • 2010
  • This study was aimed to clerify the cytotoxicity of some plant extracts such as Hosta longissima HONDA (HL), Hemerocallis fulva var. Kwanso REGL (HFVK), Hemerocallis fulva L (HF), Macrocapium officinale NAKAI (MO) and Mentha canadensis var. piperascens HARA (MCVP), the cultured NIH3T3 fibroblasts were treated with 25, 50, 100, 150 and $200{\mu}g/mL$ of five kinds of plant extracts for 48 hours, respectively. The cytotoxicity of plant extracts was measured by MTT and NR assays for the cell viability, and XTT assay for the cell adhesion activity. In this study, HL, MO and FHVK extracts showed the range of midtoxic-non toxic by the criteria of chemical cytotoxicity. While, the HF and MCVP extracts showed midtoxic. In the extract cytotoxicity, HL, MO and FHVK extracts showed non-toxic by the criteria of extract cytotoxicity. While, HF extract was determined as lower-toxic. In the responsive sensitivity of each plant extract on colorimetric assays, HF extract was sensitive to mitochondrial enzyme by MTT assay, lysosomal enzyme by NR assay and mitochondrial nucleus by XTT assay. While, MCVP extract was sensitive to mitochondrial enzyme by MTT assay and lysosomal enzyme by NR assay than other assays. While, HL, HFVK and MO extracts were most sensitive to NR assay. Cell culture is one of useful materials in the screening of cytotoxic and recovary effect on the putative chemical agents or plant extract. And also, colorimetric assay is regarded as very useful tools for quantitative measurement of cytotoxic effect on plant extracts in vitro.

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Hydrothermal Synthesis of $TiO_2$ Nanowire Array for Osteoblast Adhesion

  • Yun, Young-Sik;Kang, Eun-Hye;Hong, Min-Eui;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.275-275
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    • 2013
  • Osteoblast is one of cells related with osseointegration and many research have conducted the adhesion of osteoblast onto the surface of implant. In the osseointegration, biocompatibility of the implant and cell adhesion to the surface are important factors. The researches related to cell adhesion have a direction from micro-scaled surface roughness to nano-scaled surface roughness with advancing nanotechnology. A cell reacts and sense to stimuli from extracellular matrix (ECM) and topography of the ECM [1]. Thus, for better osseointegration, we should provide an environment similar to ECM. In this study, we synthesize TiO2 nanowires using hydrothermal reaction because TiO2 provides inertness to titanium on its surface and enables it used as an implant material for the orthopedic treatment such as fixation of the bone fracture [2]. Ti substrate is immersed into NaOH aqueous solution. The solution are heated at $140{\sim}200^{\circ}C$ for various time (10~720 minutes). After heat treatment, we take out the sample and immerse it into HCl aqueous solution for 1 hour. The acid treated sample is heated again at $500^{\circ}C$ for 3 hours [3]. Then, we culture osteoblast on the TiO2 nanowires. For investigating cell adhesion onto nanostructured surface, we conduct several tests such as MTT assay, ALP (Alkaline phosphatase) activity assay, measuring calcium expression, and so on. These preliminary results of the cell culture on the nanowires are foundation for investigating cell-material interaction especially with nanostructure interaction.

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Surface Immobilization of Amphiphilic Comb-like Polymer on Polydimethylsiloxane and in vitro Cytotoxicity Assay (양친성 빗 모양 고분자의 PDMS 표면 고정화 및 세포독성 평가)

  • Choi, Jaeyoo;Jung, Jaeyeon;Cheng, Jie;Lee, Jonghwan;Hyun, Jinho;Kim, Hyunjoong
    • Journal of Adhesion and Interface
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    • v.11 no.2
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    • pp.70-75
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    • 2010
  • It described the modification of polydimethylsiloxane (PDMS) with amphiphilic methyl methacrylate-based polyethylene glycol (PMMA-b-PEG) to enhance the hydrophilicity of a PDMS surface and cytotoxicity of it. PMMA-b-PEG solutions in water/ethanol mixture was spun-cast on the PDMS surface and the surface was characterized by long-term measurement of water contact angle. The morphology of PDMS surfaces coated with PMMA-b-PEG was characterized by field emission scanning electron microscopy and atomic force microscope. Cytotoxicity of the modified surfaces was investigated by MTT assay which would be necessary for the evaluation of tissue compatibility after implantation of the materials. Based on the MTT assay, PDMS coated with PMMA-b-PEG didn't show any significant cytotoxcity.

Inhibitory effect of the extract of Catalpa ovata G. Don. on endothelial adhesion molecule expression (개오동나무 추출물의 내피세포 부착분자 발현 억제 효과)

  • Choi, Byung-Min;Chong, Myong-Soo;Song, Ho-Joon
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.137-143
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    • 2007
  • Objectives : Catalpa ovata G. Don (Bignoniaceae) has been shown to possess a variety of pharmacological activities. However, the effect of Catalpa ovata G. Don on endothelial adhesion molecule expression has not been reported. Methods : To examine the effect of Catalpa ovata G. Don on the expression of adhesion molecules in human umbilical vein endothelial cells (HUVECs) stimulated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), we used various methods such as Western blot analysis, reverse tranascription-polymerase chain reaction (RT-PCR), and luciferase activity assay. Results : 1. The extract of Catalpa ovata G. Don inhibited the expression of intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in HUVECs stimulated with TNF-${\alpha}$. 2. The extract of Catalpa ovata G. Don reduced TNF-${\alpha}$-induced adhesion of leukocytes to HUVECs. 3. In addition, The extract of Catalpa ovata G. Don inhibited the promoter activities of ICAM-1 and VCAM-1. Conclusions : These results that Catalpa ovata G. Don may be beneficial in the treatment of inflammatory such as atherosclerosis.

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Modulatory Effect of Diethylstilbestrol on CD29-Mediated Cell-cell Adhesion in Monocytic U937 Cells (Diethylstilbestrol의 단핵구의 세포간 유착과정 조절효과)

  • Kim, Byung-Hun;Cho, Jae-Youl
    • YAKHAK HOEJI
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    • v.52 no.2
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    • pp.111-116
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    • 2008
  • Diethylstilbestrol (DESB) is a synthetic estrogen not only that routinely prescribed, but also that known to be a teratogen. In this study, we found a novel pharmacological feature that DESB is able to positively modulate CD29 $({\beta}1-integrin)$ function. Thus, DESB up-regulated homotypic cell-cell adhesion of monocytic U937 cells mediated by CD29. However, DESB did not increase the surface level of CD29 and its binding activity to ligand (fibronectin), according to flow cytometric analysis and cell-fibronectin adhesion assay. Instead, the DESB-mediated up-regulation of cell-cell adhesion was blocked by several signaling enzyme inhibitors. Treatment of U0126 [an extracellular signal-regulated kinase (ERK) inhibitor], SB20358 (a p38 inhibitor) or Rp-8-pCPT-cGMP (a protein kinase G inhibitor) clearly inhibited DESB-mediated up-regulation of cell-cell adhesion induced by CD29. However, estrogen receptor antagonist ICI 182,780 failed to abrogate DESB effect. Therefore, our data suggest that DESB may up-regulate CD29-mediated cell-cell adhesion via modulating intracellular signaling enzymes such as ERK, PKG, and p38, independent of estrogen receptor function.

Adhesion Properties of Indigenous Dadih Lactic Acid Bacteria on Human Intestinal Mucosal Surface

  • Dharmawan, Jorry;Surono, Ingrid S.;Kun, Lee Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.751-755
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    • 2006
  • Dadih is Indonesian traditional fermented buffalo milk believed by the natives to have beneficial effects on human health. This may be due to the probiotic properties possessed by the lactic acid bacteria (LAB) involved in its fermentation process. It was discovered that ten strains of dadih lactic isolates possessed some probiotic properties in vitro. In this study, the adhesion properties of dadih LAB, in comparison with documented probiotic strains, were investigated in vitro by using mucin extracted from human faeces and Caco-2 cells as the models for human intestinal mucosal surface and intestinal cells respectively. The adhesion results showed the distinction of Lactobacillus reuteri IS-27560 in adhering to both mucus layer and Caco-2 cells. The competition assay for adhesion to the mucus layer between dadih LAB and selected pathogens indicated the competence of Lactococcus lactis IS-16183 and Lactobacillus rhamnosus IS-7257 in significantly inhibiting the adhesion of Escherichia coli O157:H7. Accordingly, these two strains may be potential candidates for use as probiotic strains. Overall, the adhesion properties of all dadih LAB strains were relatively comparable to that of Lactobacillus casei Shirota and Lactobacillus rhamnosus GG, the documented probiotic strains.

Delphinidin Chloride Effects on the Expression of TNF-$\alpha$ Induced Cell Adhesion Molecules (TNF-$\alpha$에 의해 유도된 세포부착분자의 발현에 대한 Delphinidin chloride의 억제 효과)

  • Koh, Eun-Gyeong;Chae, Soo-Chul;Seo, Eun-Sun;Na, Myung-Suk;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.88-94
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    • 2009
  • The process of atherosclerosis begins through secretion of inflammatory cytokine or adhesion of leukocyte from damage in blood vessels and transmigration. This study was conducted to investigate the effects of delphinidin chloride (DC) in the initial process of atherosclerosis on the expression of ICAM-1 (Intracellular Adhesion Molecule-1) and VCAM-1 (Vascular Adhesion Molecule-1) related to adhesion of leukocyte at the HUVEC (human umbilical vein endothelial cell line. As a result, cell growth inhibition rate at 50 ${\mu}M$ was respectively 4, 3 and 5% without cell toxicity. As a result of morphological observation monocyte-endothelial cell adhesion assay and optical microscope carried out to measure attachment of mononuclear cells to endothelial cells induced by Tumor necrosis factor-alpha (TNF-$\alpha$) at concentrations without cell toxicity, DC concentration-dependently suppressed attachment. When effects on the expression of VCAM-1 and ICAM-1, cell adhesion molecules induced from endothelial cells by TNF-$\alpha$, were comparatively analyzed using western blot analysis and RT-PCR methods, protein of VCAM-1 and ICAM-1 and expression at the level of mRNA were concentration-dependently reduced. Taken together, the results of this studies provide evidence that DC possess an anti-metastatic activity.