• Title/Summary/Keyword: Adhesion assay

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Adhesion of Human Intervertebral Disk Cells on Aiginate/PLGA Microspheres (Alginate/PLGA 미립구에 대한 인간디스크 세포 부착 효과)

  • Lee, Jun-Hee;Jang, Ji-Wook;So, Jeong-Won;Choi, Jin-Hee;Park, Jong-Hak;Ahn, Shik-Il;Son, Young-Suk;Min, Byoung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.7-12
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    • 2009
  • PLGA microspheres have been known as an injectable system for tissue engineering. The purpose of this study was to investigate the condition of emulsion formation and cell adhesion on the microsphere surface. BSA-loaded PLGA microsphere was fabricated by oil-in-water (O/W) and water-in-oil-in-water (W/O/W) solvent evaporation method. Sodium alginate was dissolved in water phase to control initial burst release and to improve lag time by PLGA bulk degradation. In addition, the morphology of cells attached on the micro spheres was studied using a scanning electron microscopy (SEM). Cellular proliferation behavior of human disc cells cultivated on PLGA micro spheres was analyzed using a MTT assay. MTT assay revealed that the cells can attach and proliferate on PLGA microspheres. According to these results, we concluded that BSA -loaded alginate/PLGA microspheres can be used as an injectable system for tissue engineering application.

A Comparison of the Effects of Silica and Hydroxyapatite Nanoparticles on Poly(ε-caprolactone)-Poly(ethylene glycol)-Poly(ε-caprolactone)/Chitosan Nanofibrous Scaffolds for Bone Tissue Engineering

  • Hokmabad, Vahideh Raeisdasteh;Davaran, Soodabeh;Aghazadeh, Marziyeh;Alizadeh, Effat;Salehi, Roya;Ramazani, Ali
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.735-750
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    • 2018
  • BACKGROUND: The major challenge of tissue engineering is to develop constructions with suitable properties which would mimic the natural extracellular matrix to induce the proliferation and differentiation of cells. Poly(${\varepsilon}$-caprolactone)-poly(ethylene glycol)-poly(${\varepsilon}$-caprolactone) (PCL-PEG-PCL, PCEC), chitosan (CS), nano-silica ($n-SiO_2$) and nano-hydroxyapatite (n-HA) are biomaterials successfully applied for the preparation of 3D structures appropriate for tissue engineering. METHODS: We evaluated the effect of n-HA and $n-SiO_2$ incorporated PCEC-CS nanofibers on physical properties and osteogenic differentiation of human dental pulp stem cells (hDPSCs). Fourier transform infrared spectroscopy, field emission scanning electron microscope, transmission electron microscope, thermogravimetric analysis, contact angle and mechanical test were applied to evaluate the physicochemical properties of nanofibers. Cell adhesion and proliferation of hDPSCs and their osteoblastic differentiation on nanofibers were assessed using MTT assay, DAPI staining, alizarin red S staining, and QRT-PCR assay. RESULTS: All the samples demonstrated bead-less morphologies with an average diameter in the range of 190-260 nm. The mechanical test studies showed that scaffolds incorporated with n-HA had a higher tensile strength than ones incorporated with $n-SiO_2$. While the hydrophilicity of $n-SiO_2$ incorporated PCEC-CS nanofibers was higher than that of samples enriched with n-HA. Cell adhesion and proliferation studies showed that n-HA incorporated nanofibers were slightly superior to $n-SiO_2$ incorporated ones. Alizarin red S staining and QRT-PCR analysis confirmed the osteogenic differentiation of hDPSCs on PCEC-CS nanofibers incorporated with n-HA and $n-SiO_2$. CONCLUSION: Compared to other groups, PCEC-CS nanofibers incorporated with 15 wt% n-HA were able to support more cell adhesion and differentiation, thus are better candidates for bone tissue engineering applications.

The High Expressed Serum Soluble Neural Cell Adhesion Molecule, a High Risk Factor Indicating Hepatic Encephalopathy in Hepatocelular Carcinoma Patients

  • Liu, Tian-Hua;Guo, Kun;Liu, Ri-Qiang;Zhang, Shu;Huang, Zhuo-Hui;Liu, Yin-Kun
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3131-3135
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    • 2015
  • Objective: To investigate whether the expression of serum soluble neural cell adhesion molecule (sNCAM) is associated with hepatic encephalopathy (HE) in hepatocelular carcinoma (HCC) patients. Materials and Methods: The Oncomine Cancer Microarray database was used to determine the clinical relevance of NCAM expression in different kinds of human cancers. Sera from 75 HCC cases enrolled in this study were assessed for expression of sNCAM by enzyme linked immunosorbent assay (ELISA). Results: Dependent on the Oncomine Cancer Microarray database analysis, NCAM was down regulated in 10 different kinds of cancer, like bladder cancer, brain and central nervous system cancer, while up-regulated in lung cancer, uterine corpus leiomyoma and sarcoma, compared to normal groups. Puzzlingly, NCAM expression demonstrated no significant difference between normal and HCC groups. However, we found by quantitative ELISA that the level of sNCAM in sera from HCC patients with HE ($347.4{\pm}151.9ng/ml$) was significantly more up-regulated than that in HCC patients without HE ($260.3{\pm}104.2ng/ml$), the p-value being 0.008. sNCAM may be an important risk factor of HE in HCC patients, the correlation coefficients was 0.278 (P<0.05) on rank correlation analysis. Conclusions: This study highlights that up-regulated level of serum sNCAM is associated with HE in HCC patients and suggests that the high expression can be used as an indicator.

Antibacterial Activity of Curcuma longa against Methicillin-resistant Staphylococcus aureus

  • You Yong Ouk;Yu Hyeon Hee;Jeon Byung Hun;Jeong Seung Il;Cha Jung Dan;Kim Shin Moo;Kim Kang Ju
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.574-579
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    • 2003
  • Methicillin-resistant Staphylococcus aureus (MRSA) has been emerging worldwide as one of the most important hospital and community pathogens. Therefore, new agents are needed to treat the MRSA. In the present study, we investigated antimicrobial activity of ethyl acetate, methanol, and water extracts of Curcuma longa L. (C. longa) aganist clinical isolates of MRSA. The ethyl acetate extract of C. long a demonstrated a higher antibacterial activity than the methanol extract or water extract. Since the ethyl acetate extract was more active than other extracts, we examined whether ethyl acetate extract may restore the antibacterial activity of β-lactams and alter the adhesion and invasion of MRSA to human mucosal fibroblasts (HMFs). In the checkerboard test, ethyl acetate extract of C. longa markedly lowered the MICs of ampicillin and oxacillin against MRSA. In the bacterial adhesion and invasion assay, MRSA intracellular invasion were notably decreased in the presence of 0.125 - 2 mg/ml of C. longa extract compared to the control group. These results suggest that ethyl acetate extract of C. longa may have antibacterial activity and the potential to restore the effectiveness of β-lactams against MRSA, and inhibit the MRSA adhesion and invasion to HMFs.

Effects of Antioxidant on the Hypoxia-induced Expression of ICAM-1 in Cultured Human Synovial Fibroblasts (저산소증에 의한 활막 섬유모세포의 ICAM-1 발현에 대한 항산화제의 영향)

  • Kim, Jung Ryul;Yoo, Wan Hee
    • IMMUNE NETWORK
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    • v.2 no.1
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    • pp.25-34
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    • 2002
  • Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia and joint destruction. The synovial fibroblasts express cell adhesion molecules and have a role in adhesive interation with inflammatory cells in synovial tissue. It has been suggested that hypoxic conditioins are thought to exist in arthritic joints, and several studies indicate that reactive oxygen species (ROS) produced in hypoxic condition can initiate events that lead to pro-adhesive changes via increased expression of adhesion molecules. So, this study wsa designed to examine whether antioxidant can inhibit hypoxia-induced expression of ICAM-1 in cultured human synovial fibroblasts. Methods: Synovial fibroblasts were isolated from synovial tissue in patients with RA and cultured at hypoxic condition. Antioxidant, PDTC (pyrrolidine dithiocarbamate) were pre-treated for an hour before the hypoxic culture and synovial fibroblasts were harvested at 0, 6, 12, 24, 48 hours time points. Cell surface ICAM-1 expression in synovial fibroblasts was examined by the flow cytometric analysis. To analyse the expression of ICAM-1 mRNA, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. The levels of cytokines in culture supernatants were measured by ELISA, and activation of NF-${\kappa}B$ was analysed by electrophoretic mobility shift assay. The adhesive reaction between synovial fibroblasts and lymphocytes was assayed by measurement of fluorescent intensity of BCECF-AM in lymphocytes. Results: Hypoxic stimuli up-regulated the ICAM-1 expression as well as the adhesive interaction of human synvial fibroblasts to lymphocytes in a time-dependent manner, and PDTC inhibited hpyoxia-induced ICAM-1 expression and cell-cell interaction. PDTC also inhibited the hypoxia-induced activation of intracellular transcription factor, NF-${\kappa}B$. PDTC decreased the amount of hypoxia-induced production of IL-$1{\beta}$ and TNF-${\alpha}$. Conclusion: These studies demonstrate that PDTC inhibit the hypoxia-induced expression of the adhesion molecule, ICAM-1 and activation of NF-${\kappa}B$ in cultured human synovial fibroblasts.

Effects of troxerutin on vascular inflammatory mediators and expression of microRNA-146a/NF-κB signaling pathway in aorta of healthy and diabetic rats

  • Che, Xing;Dai, Xiang;Li, Caiying
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.5
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    • pp.395-402
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    • 2020
  • This study has investigated the effect of a potent bioflavonoid, troxerutin, on diabetes-induced changes in pro-inflammatory mediators and expression of microRNA-146a and nuclear factor-kappa-B (NF-κB) signaling pathway in aortic tissue of type-I diabetic rats. Male Wistar rats were randomly divided into four groups (n = 6/each): healthy, healthy-troxerutin, diabetic, and diabetic-troxerutin. Diabetes was induced by streptozotocin injection (60 mg/kg; intraperitoneally) and lasted 10 weeks. Troxerutin (150 mg/kg/day) was administered orally for last month of experiment. Inflammatory cytokines IL-1β, IL-6, and TNF-α, as well as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), cyclooxygenase-II (COX-II), and inducible-nitric oxide synthase (iNOS) were measured on aortic samples by enzyme-linked immunosorbent assay. Gene expressions for transcription factor NF-κB, interleukin-1 receptor-associated kinase-1 (IRAK-1), TNF receptor-associated factor-6 (TRAF-6), and microRNA-146a were determined using real-time polymerase chain reaction. Ten-week diabetes significantly increased mRNA levels of IRAK-1, TRAF-6, NF-κB, and protein levels of cytokines IL-1β, IL-6, TNF-α, adhesion molecules ICAM-1, VCAM, and iNOS, COX-II, and decreased expression of microRNA-146a as compared with healthy rats (p < 0.05 to p < 0.01). However, one month treatment of diabetic rats with troxerutin restored glucose and insulin levels, significantly decreased expression of inflammatory genes and pro-inflammatory mediators and increased microRNA level in comparison to diabetic group (p < 0.05 to p < 0.01). In healthy rats, troxerutin had significant reducing effect only on NF-κB, TNF-α and COX-II levels (p < 0.05). Beside slight improvement of hyperglycemia, troxerutin prevented the activation of NF-κB-dependent inflammatory signaling in the aorta of diabetic rats, and this response may be regulated by microRNA-146a.

In vitro evaluation of a removable partial denture framework using multi-directionally forged titanium

  • Suzuki, Ginga;Shimizu, Satoshi;Torii, Mana;Tokue, Ai;Ying, Guo;Yoshinari, Masao;Hoshi, Noriyuki;Kimoto, Katsuhiko;Miura, Hiromi;Hayakawa, Tohru;Ohkubo, Chikahiro
    • The Journal of Advanced Prosthodontics
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    • v.12 no.6
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    • pp.369-375
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    • 2020
  • PURPOSE. This study evaluated the availability of multi-directionally forged (MDF) titanium (Ti) as a component of removable partial dentures (RPDs). MDF-Ti remarkably improved the mechanical properties of RPDs due to its ultrafine-grained structure. MATERIALS AND METHODS. The wear resistance, plaque adhesion, and machinability of MDF-Ti were tested. As controls, commercially pure (CP) titanium was used for wear, plaque adhesion, and machinability tests. For wear resistance, the volume losses of the titanium teeth before and after wear tests were evaluated. Plaque adhesion was evaluated by the assay of Streptococcus mutans. In the machinability test, samples were cut and ground by a steel fissure bur and carborundum (SiC) point. An unpaired t-test was employed for the analysis of the significant differences between MDF-Ti and the control in the results for each test. RESULTS. Wear resistance and plaque adherence of MDF-Ti similar to those of CP-Ti (P>.05) were indicated. MDF-Ti exhibited significantly larger volume loss than CP-Ti in all conditions except 100/30,000 g/rpm in machinability tests (P<.05). CONCLUSION. Although the wear resistance and plaque adherence of MDF-Ti were comparable to those of controls, MDF-Ti showed better machinability than did CP-Ti. MDF-Ti could be used as a framework material for RPDs.

Modulation of Adhesion Proteins Integrin β1 and FAK, and Cytoskeletal Protein Actin by Spermine in MCF-7 Cells (MCF-7 세포에서 spermine에 의한 부착단백질 Integrin β1과 FAK, 세포골격 단백질 actin의 조절)

  • Jee, Hye-Jin;Kim, Byeong-Gee
    • Journal of Life Science
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    • v.22 no.1
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    • pp.16-24
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    • 2012
  • Polyamines are essential for cell growth and differentiation; however their precise roles are unclear yet. In the present study, the cytotoxic effect of spermine (spm) on MCF-7 cells was investigated. In the MTT assay of MCF-7 cells treated with spm, cell viability was significantly decreased in a time-and dose-dependent manner. Cell viability measurement was confirmed by trypan blue staining. FACS analysis shows that sub-G1 was increased in a time-and dose-dependent manner too. When the cells were treated with spm, cells started to show morphological changes within 2 hrs. The expression of adhesion proteins (FAK and integrin ${\beta}1$), and cytoskeletal protein (actin) was checked by Western blotting analysis. Integrin ${\beta}1$ levels were slightly decreased, and FAK and actin levels were rapidly decreased with spm treatment. In confocal laser scanning microscopy, the distribution of actin did not change but the expression decreased in a dose-dependent manner with spm treatment. FAK was evenly distributed under the plasma membrane in the untreated control. However, at 10 ${\mu}M$ spm FAK seemed to move toward the cell nucleus. Integrin ${\beta}1$, which was mainly found in the focal point of the plasma membrane in the untreated control, dispersed through the entire plasma membrane in spm treatment. The present results indicate that cytotoxic effects of spm are triggered by the disruption of adhesion proteins and cytoskeletal protein.

Effects of Antibacteria and Adhesive Inhibition of Scutellaria baicalensis Extract on Streptococcus mutans (황금(Scutellaria baicalensis) 추출물에 의한 Streptococcus mutans의 항균 및 부착억제 효과)

  • Paek, Jong-Yoon;Kim, Young-Hyun;Kwon, Hyun-Jeoung;Kim, Eun-Nim;Kim, Wan-Jong;Han, Man-Deuk
    • Journal of dental hygiene science
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    • v.8 no.4
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    • pp.367-373
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    • 2008
  • The natural products are used to be development of new antibacterial substances against human pathogenic bacteria. Adherence to the tooth surface by S. mutans is an important step in initiation of dental caries. This study was to examine antibacterial activity and anti-adhesive effect of Scutellaria baicalensis extract against S. mutans. Extracts of S. baicalensis were tested for antimicrobial activities by paper disc methods and radial diffusion assay methods, and bacterial adherence assay using 3 type of hydroxyapatite. The antibacterial level of ethyl acetate extract, IPK-3 on the growth of S. mutans was 125 mg/ml of minimum inhibitory concentration (MIC). The maximum growth of S. mutans in medium added with IPK-3 extract (50 mg/ml) was delayed to 30 hr, while the highest at 24 hr in control medium. The pH values of the control medium was 5.63 at 18 hr, but the media supplemented with IPK-3 extract was pH 6.50 at 12 hr. In adhesive inhibition assay, S. mutans was labelled with the fluorescent indicator DAPI and measured with fluorescence microscope. Adhesion of S. mutans on hydroxyapatite beads was inhibited by IPK-3 extracts. These results suggest that S. baicalensis extract can be used as an effective material for antibacterial activity and adhesive inhibition against S. mutans.

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Probiotic Potential of Pediococcus acidilactici and Enterococcus faecium Isolated from Indigenous Yogurt and Raw Goat Milk

  • Sarkar, Shovon Lal;Hossain, Md. Iqbal;Monika, Sharmin Akter;Sanyal, Santonu Kumar;Roy, Pravas Chandra;Hossain, Md. Anwar;Jahid, Iqbal Kabir
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.276-286
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    • 2020
  • Probiotics are live microorganisms that, when administered in adequate amounts, confer health benefits to the host. This study was conducted for the isolation of potential lactic acid bacteria (LAB) with probiotic properties from goat milk and yogurt. Several tests were conducted in vitro using the standard procedures for evaluating the inhibitory spectra of LAB against pathogenic bacteria; tolerance to NaCl, bile salt, and phenol; hemolytic, milk coagulation, and bile salt hydrolase activities; gastrointestinal transit tolerance; adhesion properties; and antibiotic susceptibility. Among 40 LAB strains screened according to culture characteristics, five isolates exhibited antagonistic properties. Three were identified as Pediococcus acidilactici, and two were identified as Enterococcus faecium, exploiting 16S rRNA gene sequencing. All the isolates succeeded in the gastrointestinal transit tolerance assay and successively colonized mucosal epithelial cells. Based on the results of these in vitro assays, both P. acidilactici and E. faecium can be considered as potential probiotic candidates.