• Title/Summary/Keyword: Adhesion assay

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Investigation of the effect of water extract from Cudrania tricuspidata fruit on tight junction in human keratinocyte (꾸지뽕 열매 추출물이 인간 유래 각질 형성 세포의 밀착연접에 미치는 영향)

  • Lee, Sang Soo;Choi, Sun Kyung;Kim, Jae Whan;Han, Hyo Sang;Kim, Kee Kwang
    • The Korea Journal of Herbology
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    • v.34 no.1
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    • pp.99-107
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    • 2019
  • Objectives : Cudrania tricuspidata (C. tricuspidata) is well-known traditional herbal remedy and its root, leaf and fruit were used for treatment of inflammation, tumor and painkilling. However, effect of C. tricuspidata fruit on tight junction is still unknown. The aim of this research was to determine the effect of C. tricuspidata fruit extract on human keratinocyte HaCaT cells. Methods : The antioxidant effects of water extract of C. tricuspidata (WECT) and ethanol extract of C. tricuspidata (EECT) were analyzed by using an ABTS assay. To confirm the cytotoxicity of WECT and EECT, MTS assay was performed. The mRNA expression levels of tight junction related genes were analyzed using quantitative RT-PCR analysis. Furthermore, dispase assay was used to investigate the alteration of cell-cell adhesion strength of EECT treated HaCaT cells. Results : WECT and EECT showed strong antioxidant activity. No obvious cytotoxicity was observed in both WECT and EECT until $2.0mg/m{\ell}$ concentration. The mRNA expression level of Claudin 6 were significantly increased by EECT treatment, whereas the WECT did not affect the expression of Claudin 6. Furthermore, EECT treatment enhances cell-cell adhesion strength. Conclusions : In this study, we investigated the physiological activities of the extracts of Cudrania tricuspidata fruit extracts on human keratinocytes by two different extraction methods. EECT might have an anti-aging activity on the skin by reducing oxidative stress. Moreover, it may be a useful ingredient in atopic dermatitis and skin-moisturizing, given its effects of altering Claudin 6 gene expression and enhancing cell-cell adhesion strength.

Effects of lycopene on number and function of human peripheral blood endothelial progenitor cells cultivated with high glucose

  • Zeng, Yao-Chi;Mu, Gui-Ping;Huang, Shu-Fen;Zeng, Xue-Hui;Cheng, Hong;Li, Zhong-Xin
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.368-376
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    • 2014
  • BACKGROUND/OBJECTIVES: The objectives of this study were to investigate the effects of lycopene on the migration, adhesion, tube formation capacity, and p38 mitogen-activated protein kinase (p38 MAPK) activity of endothelial progenitor cells (EPCs) cultivated with high glucose (HG) and as well as explore the mechanism behind the protective effects of lycopene on peripheral blood EPCs. MATERIALS/METHODS: Mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation. EPCs were identified after induction of cellular differentiation. Third generation EPCs were incubated with HG (33 mmol/L) or 10, 30, and $50{\mu}g/mL$ of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation and apoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performed by replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity was assayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylated and non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and $50{\mu}g/mL$ of lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantly lower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirm the causal relationship between MAPK inhibition and the protective effects of lycopene against HG-induced cellular injury, we treated cells with SB203580, a phosphorylation inhibitor. The inhibitor significantly inhibited HG-induced EPC injury. CONCLUSIONS: Lycopene promotes proliferation, migration, adhesion, and in vitro vasculogenesis capacity as well as reduces apoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-induced EPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-induced EPC injury by inhibiting p38 MAPKs.

Steroidal Saponins from Paris polyphylla Suppress Adhesion, Migration and Invasion of Human Lung Cancer A549 Cells Via Down-Regulating MMP-2 and MMP-9

  • He, Hao;Zheng, Lei;Sun, Yan-Ping;Zhang, Guang-Wei;Yue, Zheng-Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10911-10916
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    • 2015
  • Background: Tumor metastases are the main reasons for oncotherapy failure. Paris polyphylla (Chinese name: Chonglou) has traditionally been used for its anti-cancer actions. In this article, we focus on the regulation of human lung cancer A549 cell metastases and invasion by Paris polyphylla steroidal saponins (PPSS). Materials and Methods: Cell viability was evaluated in A549 cells by MTT assay. Effects of PPSS on invasion and migration were investigated by wound-healing and matrigel invasion chamber assays. Adhesion to type IV collagen and laminin was evaluated by MTT assay. Expression and protease activity of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, were analyzed by Western blotting and gelatin zymography, respectively. Results: PPSS exerted growth inhibitory effects on A549 cells, and effectively inhibited A549 cell adhesion, migration and invasion in a concentration-dependent manner. Western blotting and gelatin zymography analysis revealed that PPSS inhibited the expression and secretion of MMP-2 and MMP-9 in A549 cells. Conclusions: PPSS has the potential to suppress the migration, adhesion and invasion of A549 cells. PPSS could be a potential candidate for interventions against lung cancer metastases.

Focal adhesion linker proteins expression of fibroblast related to adhesion in response to different transmucosal abutment surfaces

  • Moon, Yeon-Hee;Yoon, Mi-Kyeong;Moon, Jung-Sun;Kang, Jee-Hae;Kim, Sun-Hun;Yang, Hong-Seo;Kim, Min-Seok
    • The Journal of Advanced Prosthodontics
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    • v.5 no.3
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    • pp.341-350
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    • 2013
  • PURPOSE. To evaluate adherence of human gingival fibroblasts (HGFs) to transmucosal abutment of dental implant with different surface conditions with time and to investigate the roles of focal adhesion linker proteins (FALPs) involved in HGFs adhesion to abutment surfaces. MATERIALS AND METHODS. Morphologies of cultured HGFs on titanium and ceramic discs with different surface were observed by scanning electron microscopy. Biocompatibility and focal adhesion were evaluated by ultrasonic wave application and cell viability assay. FALPs expression levels were assessed by RT-PCR and western blot. RESULTS. There seemed to be little difference in biocompatibility and adhesion strength of HGFs depending on the surface conditions and materials. In all experimental groups, the number of cells remaining on the disc surface after ultrasonic wave application increased more than 2 times at 3 days after seeding compared to 1-day cultured cells and this continued until 7 days of culture. FALPs expression levels, especially of vinculin and paxillin, also increased in 5-day cultured cells compared to 1-day cultured fibroblasts on the disc surface. CONCLUSION. These results might suggest that the strength of adhesion of fibroblasts to transmucosal abutment surfaces increases with time and it seemed to be related to expressions of FALPs.

Loquat (Eriobotrya japonica) extracts suppress the adhesion, migration and invasion of human breast cancer cell line

  • Kim, Min-Sook;You, Mi-Kyoung;Rhuy, Dong-Young;Kim, Yung-Jae;Baek, Hum-Young;Kim, Hyeon-A
    • Nutrition Research and Practice
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    • v.3 no.4
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    • pp.259-264
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    • 2009
  • We examined the inhibitory effects of loquat methanol extract on the adhesion, migration, invasion and matrix metalloproteinase (MMP) activities of MDA-MB-231 human breast cancer cell line. Cells were cultured with DMSO or with 10, 25, or 50 ${\mu}g/ml$ of loquat methanol extract. Both leaf and seed extracts significantly inhibited growth of MDA-MB-231 cells in a dose-dependent manner, although leaf extract was more effective. Adhesion and migration were significantly inhibited by loquat extracts in a dose-dependent manner. Loquat extract also inhibited the invasion of breast cancer cells in a dose-dependent manner and leaf extract was more effective than seed extract. MMP-2 and MMP-9 activities were also inhibited by loquat extract. Our results indicate that methanol extracts of loquat inhibit the adhesion, migration and invasion of human breast cancer cells partially through the inhibition of MMP activity and leaf extract has more anti-metastatic effects in cell based assay than seed extract. Clinical application of loquat extract as a potent chemopreventive agent may be helpful in limiting breast cancer invasion and metastasis.

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.

Evaluation of C. Albicans and S. Mutans adherence on different provisional crown materials

  • Ozel, Gulsum Sayin;Guneser, Mehmet Burak;Inan, Ozgur;Eldeniz, Ayce Unverdi
    • The Journal of Advanced Prosthodontics
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    • v.9 no.5
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    • pp.335-340
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    • 2017
  • PURPOSE. Bacterial adhesion on provisional crown materials retained for a long time can influence the duration for which permanent prosthetic restorations can be healthily worn in the oral cavity. The aim of this study was to compare seven different commonly used provisional crown materials with regard to Streptococcus mutans and Candida albicans surface adhesion. MATERIALS AND METHODS. For each group, twenty specimens of the provisional fixed prosthodontic materials TemDent ($Sch{\ddot{u}}tz$), Imident (Imicryl), Tab 2000 (Kerr), Structur Premium (Voco), Systemp (Ivoclar Vivadent), Acrytemp (Zhermack), and Takilon-BBF (Takilon) were prepared (diameter, 10.0 mm; height, 2.0 mm). Surface roughness was assessed by atomic force microscopy. Each group was then divided into 2 subgroups (n=10) according to the microbial suspensions used: S. mutans and C. albicans. The specimens were incubated at $37^{\circ}C$ with S. mutans or C. albicans for seven days. Bacterial adherence on surfaces was assessed using the 2,3-bis[2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) assay. RESULTS. S. mutans showed maximum adhesion to Structur, followed by Systemp, Acrytemp, Takilon, Tab 2000, Imident, and TemDent (P<.05). The highest vital C. albicans adhesion was noted on Takilon, followed by Imident and Tab 2000; the lowest adhesion was noted on Systemp (P<.05). CONCLUSION. The materials showed significant differences in the degree of bacterial adhesion. C. albicans showed higher surface adhesion than S. mutans on provisional crown and fixed partial denture denture materials.

Study on Antitumor Activity and Immunomodulatory effects of Seoleosojong-tang (활어소종탕이 항종역반응에 미치는 영향)

  • Son Ki Jeong;Park Yang Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.1
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    • pp.137-147
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    • 2004
  • In order to evaluate the antitumor activity and immunomodulatory effects of Seoleosojong-tang(SST), studies were done. We measured the cytotoxic activity for various kinds of cancer cells, inhibitory effect on activity of DNA topoisomerase I, cell adhesion to complex extracellular matrix, survival time in ICR bearing S-180, pulmonary colonization and histological changes of lung in C57BL/6 injected i.v. with B16-F10, CAM assay, expression of CD4/sup +/, CD8/sup +/, B220/sup +/, cytokine gene in spleen cell. The results were obtained as follows: 1. In cytotoxicity against A549, HT1080, 816-F10, NCL-H661 was showed cytotoxicity as compared with control. 2. The inhibitory effect on adhesion of A549, 816-F10 to complex extracellular matrix was over 40% at 100 ㎍/㎖ of SST. 3. In DNA topoisomerase I assay, SST has inhibitory effect. 4. The T/C% was 120.8 in SST treated group in S-180 bearing ICR mice. 5. In pulmonary colonization assay, a number of colonies were decreased significantly and histological changes were showed that infiltration area of cancer cells were inhibited effectively in SST treated group. 6. In CAM Assay, SST has antiangiogenic effect. 7. On the expression of positive cell to CD4/sup +/, CD8/sup +/ and 8220/sup +/ in spleen cells, CD4/sup +/ cells were increased significantly in SST treated group. 8. Effect of SST on IL-1β gene expression in splenic cell was significantly increased as function of whole concentration. 9. The gene expression of IL-4, IL-6, IL-10, IL-12, IFN-γ, TNF-α were increased in SST treated group. From above results SST could be usefully applied for antitumor activity and immunomodulatory effects, but further research of SST should be required.

Screening of Lactic Acid Bacteria with Potent Adhesive Property in Human Colon using Colonic Mucin-binding Assay (Colonic mucin-binding assay를 이용한 장내 우수 점착능 유산균주의 선별)

  • Kim, Seong-Yeong;Shin, Kwang-Soon;Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.36 no.6
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    • pp.959-967
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    • 2004
  • To screen probiotic lactic acid bacteria with potent adhesive property on human colonic mucosa, colonic mucin-binding assay was introduced. This colonic mucin-binding assay actually measures the binding activity of surface lectin-like protein (SLP) on colonic mucin, and the optimal conditions were examined. The optimal pH for colonic mucin coating on plate wells was 4.8, and ${\times}24,000$ diluted solution of commercially available horseradish peroxidase (HRP) conjugated streptoavidin yielded good results, for rapid screening, $5.0\;{\mu}g/mL$ of biotinylated SLP from lactic acid bacteria was optimal, and optimal scintillation time of 3,3',5,5'-tetramethyl benzidine (TMB) was 10 min. These conditions were useful for both rapid selection and quantitative analysis of lactic acid bacteria that have high adhesion property to human intestinal tract. Among 50 strains of lactic acid bacteria, including 32 type culture strains and 18 isolated strains from infant feces, Lactobacillus species FSB-1 isolated from kimchi showed the highest binding activity to colonic mucin. From taxonomical viewpoints based on morphological study, physico-biochemical study, partial 16S rDNA seguencing, and phylogenetic analysis, L. species FSB-1 was identified as Lactobacillus brevis.

Bioactive characteristics of an implant surface coated with a pH buffering agent: an in vitro study

  • Pae, Hyung-Chul;Kim, Su-Kyoung;Park, Jin-Young;Song, Young Woo;Cha, Jae-Kook;Paik, Jeong-Won;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.49 no.6
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    • pp.366-381
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    • 2019
  • Purpose: The purpose of this study was to evaluate the effectiveness of conventional sandblasted, large-grit, acid-etched (SLA) surface coated with a pH buffering solution based on surface wettability, blood protein adhesion, osteoblast affinity, and platelet adhesion and activation. Methods: Titanium discs and implants with conventional SLA surface (SA), SLA surface in an aqueous calcium chloride solution (CA), and SLA surface with a pH buffering agent (SOI) were prepared. The wetting velocity was measured by the number of threads wetted by blood over an interval of time. Serum albumin adsorption was tested using the bicinchoninic acid assay and by measuring fluorescence intensity. Osteoblast activity assays (osteoblast adhesion, proliferation, differentiation, mineralization, and migration) were also performed, and platelet adhesion and activation assays were conducted. Results: In both the wetting velocity test and the serum albumin adsorption assay, the SOI surface displayed a significantly higher wetting velocity than the SA surface (P=0.000 and P=0.000, respectively). In the osteoblast adhesion, proliferation, differentiation, and mineralization tests, the mean values for SOI were all higher than those for SA and CA. On the osteoblast migration, platelet adhesion, and activation tests, SOI also showed significantly higher values than SA (P=0.040, P=0.000, and P=0.000, respectively). Conclusions: SOI exhibited higher hydrophilicity and affinity for proteins, cells, and platelets than SA. Within the limits of this study, it may be concluded that coating an implant with a pH buffering agent can induce the attachment of platelets, proteins, and cells to the implant surface. Further studies should be conducted to directly compare SOI with other conventional surfaces with regard to its safety and effectiveness in clinical settings.