• Title/Summary/Keyword: adherent cells

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Comparison of alkaline phosphatase activity of MC3T3-E1 cells cultured on different Ti surfaces: modified sandblasted with large grit and acid-etched (MSLA), laser-treated, and laser and acid-treated Ti surfaces

  • Li, Lin-Jie;Kim, So-Nam;Cho, Sung-Am
    • The Journal of Advanced Prosthodontics
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    • v.8 no.3
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    • pp.235-240
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    • 2016
  • PURPOSE. In this study, the aim of this study was to evaluate the effect of implant surface treatment on cell differentiation of osteoblast cells. For this purpose, three surfaces were compared: (1) a modified SLA (MSLA: sand-blasted with large grit, acid-etched, and immersed in 0.9% NaCl), (2) a laser treatment (LT: laser treatment) titanium surface and (3) a laser and acid-treated (LAT: laser treatment, acid-etched) titanium surface. MATERIALS AND METHODS. The MSLA surfaces were considered as the control group, and LT and LAT surfaces as test groups. Alkaline phosphatase expression (ALP) was used to quantify osteoblastic differentiation of MC3T3-E1 cell. Surface roughness was evaluated by a contact profilometer (URFPAK-SV; Mitutoyo, Kawasaki, Japan) and characterized by two parameters: mean roughness (Ra) and maximum peak-to-valley height (Rt). RESULTS. Scanning electron microscope revealed that MSLA (control group) surface was not as rough as LT, LAT surface (test groups). Alkaline phosphatase expression, the measure of osteoblastic differentiation, and total ALP expression by surface-adherent cells were found to be highest at 21 days for all three surfaces tested (P<.05). Furthermore, ALP expression levels of MSLA and LAT surfaces were significantly higher than expression levels of LT surface-adherent cells at 7, 14, and 21 days, respectively (P<.05). However, ALP expression levels between MSLA and LAT surface were equal at 7, 14, and 21 days (P>.05). CONCLUSION. This study suggested that MSLA and LAT surfaces exhibited more favorable environment for osteoblast differentiation when compared with LT surface, the results that are important for implant surface modification studies.

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.

IN VITRO STUDY ON THE ADHERENCE AND PENETRATION OF CANDIDA ALBICANS INTO DENTURE SOFT LINING MATERIALS (의치 연성이장재에 대한 Candida albicans의 부착과 침투연구)

  • Kim Min-Ju;Shin Sang-Wan;Lee Jeong-Yeol
    • The Journal of Korean Academy of Prosthodontics
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    • v.44 no.4
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    • pp.466-476
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    • 2006
  • Purpose : Colonization of denture soft lining materials by Candida albicans can result in clinical problem, and deterioration of the materials. This study aimed to compare the retention and penetration of C. albicans into four denture soft lining materials commonly used. Materials and methods : Four denture soft lining materials (Coe-comfort$^{(R)}$, Coe-soft$^{(R)}$, GC soft liner$^{(R)}$, and Tissue conditioner$^{(R)}$) discs were prepared to glass slide and dental stone. Adherence of yeast to surfaces was monitored after one hour incubation of standardized washed cell suspension with test disc surfaces. Adherent cells stained with acridine orange were counted fluorescence microscopy. Penetration of yeast into materials bonded with acrylic resin after 1, 2, 3,4, 5, 6 and 7 days incubation was observed through sections stained using acridine orange and estimated to quantitative analysis using radioisotope. Results : There was statistical significance in cell numbers between smooth and rough surfaces(p<0.05). Higher numbers of cells were observed on rough surfaces. There was statistical significance in adherent cell numbers into smooth and rough surfaces individually(p<0.05). According to the increase of incubation periods, the cells penetrated into denture soft lining materials were shown to increase. The differences among all kinds of soft liner were statistically significant(p<0.05),and the largest number of cells penetrated into soft liners was observed in the Coe-soft$^{(R)}$. Conclusion : Initial adherence and penetration of yeast into denture soft lining materials has been influenced by surface roughness and chemical composition of them. The selection of appropriate materials and their fabrication may promote clinical performance.

Heat-Shock Protein 70 as a Tumor Antigen for in vitro Dendritic Cell Pulsing in Renal Cell Carcinoma Cases

  • Meng, Fan-Dong;Sui, Cheng-Guang;Tian, Xin;Li, Yan;Yang, Chun-Ming;Ma, Ping;Liu, Yun-Peng;Jiang, You-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8947-8950
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    • 2014
  • Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

Opsonized Streptococcus mutans influenced maturation of human dendritic cells

  • Ida, N.;Ozaki, K.;Suenobu, S.;Takashi, K.;Yamaguchi, D.;Matsuo, T.
    • Proceedings of the KACD Conference
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    • 2003.11a
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    • pp.602-602
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    • 2003
  • I. Objectives It is reported that there are complements and immunogloblins in serum from dental pulp in dentinal tubules, and it is thought that dental caries bacteria is opsonized by these serum ingredients, and it is presented by dendritic cells(DCs) in dental pulp. So, we examined whether a maturational difference of DCs occured when S. mutans was opsonized. II. Materials and Methods PBMC was divided from normal human peripheral blood and collected CD14 positive cells by magnetic beads system. Adherent cells were incubated in 5% FCS-RPMI medium included GM-CSF, IL-4 for seven days.(omitted)

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The Prevalence of Enteroadherent Escherichia coli(EAEC) and the Study of its Pathogenic Role in Korean Children with Diarrhea (우리나라 소아 설사에 있어서의 Enteroadherent Escherichia coli의 병원적 역학)

  • Kim, Jung-Mogg;Kim, Kyung-Hee;Cho, Yaug-Ja;Suh, Inn-Soo
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.139-145
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    • 1987
  • Adherence to HEp-2 cells has been proposed as a virulence characteristic of enteropathogenic E. coli (EPEC). The role of the HEp-2 adherent E. coli was evaluated in a group of children with endemic diarrhea admitted to Hanyang University Hospital in Seoul, Korea. HEp-2-adherent E. coli was detected in fecal samples of 59 (59%) of 100 cases and ten (22.7%) of 44 concurrent control children (p<0.0005). Adherence was exhibited by 15 serogroups and subgroups, but within these groups more than one adherence pattern was frequently observed. Of 17 strains belonging to traditional infantile EPEC serogroups, 12(70.6%) gave a positive adherence. Of 45 enterotoxin producing strains, 24 (53.3%) gave a positive adherence. HEp-2-adherent strains that did not belong to classic EPEC serogroups and did not produce heat-stable and/or heat-labile enterotoxins(referred as enteroadherent E. coli, EAEC) was found in 29 (29%) of the patients with diarrhea and in six (13.6%) of the well children (p<0.05). From 22 of the 29 cases, no pathogen other than EAEC was isolated. These findings strongly implicate EAEC as the cause of diarrhea in the children. Our study supports the concept that EAEC may be an important cause of endemic diarrhea in Korean children.

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Screening and Isolation of the Cell Differentiation Inducers from Medicinal Plants (I) (생약으로부터 세포분화유도물질의 검색 및 분리 및 분리 (I))

  • Park, Eun-Jung;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.225-232
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    • 1997
  • 300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

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Modulation of the activity of ex vivo cultured leukemic-DC by L-ascorbic acid (LAA)

  • Lee, Hyun-Ah;Baek, So-Young;Lee, Hong-Gi;Kim, Ki-Hyun;Park, Chan-Hyung
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.311.2-311.2
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    • 2002
  • L -ascorbic acid (LAA) was shown to modulate the in vitro growth of leukemic-colony forming cells from patients with acute myelogeneous leukemia (AML). Dendritic cells (DCs) were successfully cultured from the leukemic blasts by us and others. The effects of LAA on the ex vivo cultured leukemic-DC were studied. Plastic adherent cells from the leukemic blasts were cultured with GM-CSF and IL -4 (each 103 U/$m\ell$) with or without LAA (300 ${\mu}$M) for 7 days and harvested. (omitted)

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Measuring Glutathione Regeneration Capacity in Stem Cells

  • Jihye Kim;Yi-Xi Gong;Eui Man Jeong
    • International Journal of Stem Cells
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    • v.16 no.3
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    • pp.356-362
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    • 2023
  • Glutathione (GSH) is a chief cellular antioxidant, affecting stem cell functions. The cellular GSH level is dynamically altered by the redox buffering system and transcription factors, including NRF2. Additionally, GSH is differentially regulated in each organelle. We previously reported a protocol for monitoring the real-time GSH levels in live stem cells using the reversible GSH sensor FreSHtracer. However, GSH-based stem cell analysis needs be comprehensive and organelle-specific. Hence, in this study, we demonstrate a detailed protocol to measure the GSH regeneration capacity (GRC) in living stem cells by measuring the intensities of the FreSHtracer and the mitochondrial GSH sensor MitoFreSHtracer using a high-content screening confocal microscope. This protocol typically analyses the GRC in approximately 4 h following the seeding of the cells onto plates. This protocol is simple and quantitative. With some minor modifications, it can be employed flexibly to measure the GRC for the whole-cell area or just the mitochondria in all adherent mammalian stem cells.

The virulence of Brucella abortus isolated from cattle in Korea (국내 소에서 분리한 Brucella abortus의 병원성 분석)

  • Lim, Jeong Ju;Kim, Jeong-Hwa;Kim, Dong Hyeok;Lee, Jin Ju;Kim, Dae Geun;Jun, Moo-Hyung;Kim, Sang Hun;Chang, Hong Hee;Lee, Hu Jang;Min, Won-Gi;Kim, Suk
    • Korean Journal of Veterinary Research
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    • v.51 no.1
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    • pp.15-20
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    • 2011
  • In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.