• Title/Summary/Keyword: Adhesion assay

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Effect of Ferulic Acid on Cell Viability and Cell Adhesion Activity in Normal Human Gingival Fibroblasts

  • Lee Joo-Hyun;Jin Byung-Jo;Son Il-Hong;Han Du-Seok
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.269-273
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    • 2004
  • This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferolic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferolic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 2,130.0 μM and 1,773.7 μM ferolic acid, respectively. These results suggest that ferolic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

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EFFECT OF TITANIUM SURFACE ROUGHNESS ON CELL ADHESION OF HUMAN OSTEOBLAST-LIKE CELLS (MG63)

  • Yim Soon-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.261-266
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    • 2004
  • Statement of problem. The effects of surface roughness have not or insufficiently been analyzed on earlier events such as cell adhesion though cell behavior most germane to implant performance is cell adhesion. Purpose. The purpose of this study was to evaluate cell adhesion of osteoblast-like cells (MG63) onto three types of titanium disks with varying roughness using the Elisa assay. Materials and methods. Representative disks from each group (SLA, HA, machined) were subjected to surface analysis and surface roughness was measured by the optical interferometer (Accura 2000, Intekplus Co., Seoul, Korea). Following this, MG63 cells were cultured on the titanium disks and released. Cell adhesion measurements using the Elisa assay were performed specifically at three points: after 24, 48, and 72 hours of culture. Results. Among the 3 types of surface analyzed, the SLA surface was the roughest with a Ra value of $1.114{\mu}m$ followed by HA coated surface and machined surface, consecutively. The optical density values for the SLA surface group was significantly higher than that of the machined and HA coated surface groups following 24 and 48 hours of culture. The cell culture on HA coated surface showed significantly higher values compared to the machined surface following 24, 48 and 72 hours of culture. Conclusion. The results suggest that surface treatment of titanium surfaces enhanced cell adhesion of human osteoblast-like cells (MG63).

Inhibition of Helicobacter pylori Adhesion by Acidic Polysaccharide Isolated from Artemisia capillaris

  • Woo, Jeung-S.;Ha, Byung-H.;Kim, Tae-G.;Lim, Yoon-Gho;Kim, Kyung-H.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.853-858
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    • 2003
  • Helicobacter pylori specifically adhere to host cells through a number of putative receptors and ligands, mainly based on carbohydrate-protein interactions. Polysaccharide fractions isolated from the leaves of Artemisia capillaris showed different inhibitory activities against H. pylori adhesion by using hemagglutination assay. Among these fractions, an acidic polysaccharide fraction FlA showed highly effective inhibitory activity, and its minimum inhibition concentration was 0.63 mg/ml. The inhibition results by the hemagglutination assay were consistent with those obtained by the enzymelinked glycosorbent assay, which was developed by the conjugation of horseradish peroxidase with fetuin, a sialic acid-containing glycoprotein which was specific to H. pylori adhesion. FlA contained the highest carbohydrate content among polysaccharide fractions, and no protein was detectable when further purified by gel filtration FPLC. Sugar composition analysis using GC revealed the highest amount of galacturonic acid among sugars, which suggests that FlA contains essentially acidic polysaccharides. Our data suggest that acidic polysaccharides may play an important role in the inhibition of H. pylori adhesion to host cells.

Modulatory Effect of BAY11-7082 on CD29-mediated Cell-cell Adhesion in Monocytic U937 Cells (BAY11-7082에 의한 U937 세포의 CD29-매개성 세포간 유착과정 조절 효과)

  • Kim, Byung-Hun;Cho, Jae-Youl
    • YAKHAK HOEJI
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    • v.52 no.5
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    • pp.412-417
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    • 2008
  • BAY11-7082 was initially found to be an anti-inflammatory drug with NF-${\kappa}B$ inhibitory property. In this study, we evaluated modulatory function of BAY11-7082 on U937 cell-cell adhesion induced by CD29 (${\beta}1$-integrins). BAY11-7082 strongly blocked functional activation of CD29 (${\beta}1$-integrins), as assessed by cell-cell adhesion assay. However, this compound did not block a simple activation of CD29, as assessed by cell-fibronectin adhesion assay. In particular, to understand molecular mechanism of BAY11-7082-mediated inhibition, the regulatory roles of CD29-induced actin cytoskeleton rearrangement under cell-cell adhesion and surface level of CD29 were examined using confocal and flow cytometic analysis. Interestingly, this compound strongly suppressed the molecular association of actin cytoskeleton with CD29 at cell-cell adhesion site. Moreover, BAY11-7082 also diminished surface levels of CD29 as well as its-associated adhesion molecule CD147, but not other adhesion molecules such as CD18 and CD43. Therefore, our data suggest that BAY11-7082 may be involved in regulating immune responses managed by CD29-mediated cell-cell adhesion.

Enzyme-Linked, Biotin-Streptavidin Bacterial-Adhesion Assay for Helicobacter pylori Lectin-Like Interactions with Cultured Cells

  • Murillo, Guzman;Antonia, Maria;Ascencio, Felipe
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.35-39
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    • 2001
  • A simple method for studying the lectin-like interactions between Helicobacter pylori and cultured human epithelial cell lines was developed using an enzyme-linked, biotin-streptavidin bacterial-adhesion assay. The present study suggests that this method is suitable for evaluating the participation of lectin interactions in the adhesion of H. pylori to cultured HeLa S3 and Kato III cells, both fixed and glycosidase-treated cells, as well as assessing glycoconjugated binding inhibition studies. The time-course and dose-dependent kinetics of the biotin-labeled H. pylori adhesion th the formaldehyde-fixed Hela S3 and Kato III cell lines exhibited saturation. In addition, the binding of the biotin-labeled H. pylori to the formaldehyde-fixed cultured cells was partially blocked by pre-incubation with glycoconjugates and polyclonal antibodies against a heparan sulfate binding protein from H. pylori.

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Inhibitory Effect of Agaricus Mixed Prescription on Metastasis and Tumor Formation

  • Chu Ji Hee;Lim Nak Cheol;Kim Sung Hoon;Ryu Ki Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.560-567
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    • 2003
  • Antimetastatic effects of Agaricus mixed prescription (AMP) were studied in the respect of blood-borne metastasis. For this aim, cytotoxicity against various cancer cells and normal cells, Chicken Chorioallantoic membrane (CAM) assay, cancer cell adhesion assay, platelet aggregation assay, pulmonary colonization, life span of S-180 implanted mice, and cytokine release assay were evaluated, respectively. The results were summarized as follows; AMP did not exert any cytotoxicity against all cell lines with IC50 of 25mg/ml on B16BL6. AMP disrupted formation of CAM at 1mg/ml. AMP was suppressive in adhesion assay of B16BL6. AMP also inhibited tumor induced platelet aggregation. In pulmonary colonization assay by B16BL6, the number of colonies in the lungs was significantly decreased in sample group than in control group. In animal study with S-180, the life span of AMP treated group was extended than that of control group. IL-12 was effectively increased in AMP treated group in cytokine release assay. Taken together, AMP can be possibly applied to cancer or metastasis.

Cytotoxic Effect of Syringic Acid on Human Oral Epithelioid Carcinoma Cells

  • Lee Joo-Hyun;Han Du-Suk;Jekal Seung-Joo;Lee Jae-Hyung;Kim Chong-Ho;Yoo Min;Park Seung-Taeck
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.337-341
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    • 2005
  • This study was undertaken to clerify the cytotoxic effect of syringic acid by colorimetric assay on human cancer cells. For the evaluation of cytotoxicity of syringic acid, the cell viability and cell adhesion activity of syringic acid on cancer cells, human oral epithelioid carcinoma cells were determined using by colorimetric assays such as MTT (3-[4,5­dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and XTT (2,3-bis-[2-methoxy-4-nitro-5-sulfophenyl]­2H-tetrazolium-5-caboxanilide) assay, respectively after human oral epithelioid carcinoma cells were treated with syringic acid for 48 hours. In this study, the cell viability of syringic acid on human oral epithelioid carcinoma cells showed a significant decrease by MTT assay compared with control, and also, the cell adhesion activity by XTT assay was decreased significantly in these cells after cells were treated with various concentrations of syringic acid for 48 hours. $MTT_{50}\;and\;XTT_{50}\;were\;282.3\;{\mu}M\;and\;418.8{\mu}M$ syringic acid, respectively. These results suggest that syringic acid shows midcytotoxic effect on human oral epithelioid carcinoma cells by the decreasement of the cell viability and the cell adehision activity assessed by colorimetric assay in these cultures.

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Effect of Various Factors on Early THP-1 Cell Adhesion Induced Phorbol 12-Myristate 13-Acetate (PMA) (Phorbol 12-myristate 13-acetate (PMA) 처리로 유도되는 THP-1 세포의 초기 부착에 관한 다양한 인자의 효과)

  • Jo, Yong-Sam;Shin, Ji-Hyun;Choi, Tae-Saeng
    • Journal of Life Science
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    • v.18 no.7
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    • pp.952-957
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    • 2008
  • We evaluated the effects of various factors (e.g., serum, inhibitors of protein synthesis, and cytoskeleton and protein kinases) on early PMA-induced THP-1 cell adhesion using an adhesion assay with Sulforhodamine B (SRB) staining, which was used to assess the proliferation of the attached cells. THP-1 cell adhesion to a plastic substrate was detected 1 hr after exposure to Phorbol 12-Myristate 13-Acetate (PMA) and peaked after 18 hr. At concentrations > 25 nM PMA, the level of adhesion did not change. Based on our preliminary results, we used 25 nM PMA and 5 hr of culture as standard assay conditions. Early PMA-induced cell adhesion was not affected by the presence of serum or PD 98059 in the culture medium, but was affected by the addition of PKC inhibitors and cycloheximide. In the presence of actin inhibitor with PMA, the cell adhesion increased when comparing with PMA treatment only. Thus, early PMA-induced adhesion of THP-1 cells does not require serum in the culture medium, MAP-kinase activation, or actin polymerization, but does require de novo protein synthesis and PKC activation. Our SRB-based cell adhesion assay may be used to screen other PKC inhibitors.

Norcantharidin Anti-Angiogenesis Activity Possibly through an Endothelial Cell Pathway in Human Colorectal Cancer

  • Yu, Tao;Hou, Fenggang;Liu, Manman;Zhou, Lihong;Li, Dan;Liu, Jianrong;Fan, Zhongze;Li, Qi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.499-503
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    • 2012
  • The present study was based on the unexpected discovery that norcantharidin exerted anti-angiogenesis activity when effects on growth of human colon cancer were studied. The aim was to further verify this finding and explore possible mechanisms using a tumor xenograft model in nude mice. We confirmed that norcantharidin (5 or 15 mg/kg) could inhibit angiogenesis of human colon cancer in vivo. In vitro, crossing river assay, cell adhesion assay and tube formation assay indicated that NCTD could reduce the migration, adhesion and vascular network tube formation ability of HUVECs. At the same time, the expression levels of VEGF and VEGFR-2 proteins which play important roles in angiogenesis were reduced as examined by western blotting analysis. Taken together, the results firstly showed NCTD could inhibit angiogenesis of human colon cancer in vivo, probably associated with effects on migration, adhesion and vascular network tube formation of HUVECs and expression levels of VEGF and VEGFR-2 proteins.

The Inhibitory Effects of Poncirin against Nickel Induced Cytotoxicity (Poncirin의 니켈에 대한 세포독성억제효과)

  • Yang, Seung-Jin;Kwak, Dong-Keun;Han, Du-Suk
    • The Korea Journal of Herbology
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    • v.21 no.2
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    • pp.121-127
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    • 2006
  • Objectives : Nickel is a major metal used in the nickel-chromium alloys of most orthodontic appliances, partial denture and implants. This study was carried out for the examination of the cytotoxicity on nickel sulfide in cultured NIH3T3 fibroblasts, and poncirin effect on nickel-induced cytotoxicity. Methods : Cell viability for the MTT assay and cell adhesion activity for the XTT assay. Results : The $IC_{50}$ of nickel sulfide by the MTT assay was $93.7\;{\mu}M$. Poncirin was significantly increased the cell viability and cell adhesion activity. Conclusion : Nickel was highly toxic and poncirin has the inhibitory effects against nickel induced cytotoxicity.

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