• Title/Summary/Keyword: L929

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THE CYTOTOXICITY ON L929 CELLS AND ANTIMICROBIAL EFFECT ON SEVERAL STREPTOCOCCI OF CALCIUM HYDROXIDE (수산화칼슘의 L929 세포독성 및 연쇄구균에 대한 항균효과에 관한 연구)

  • Yu, Young-Dae;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.538-548
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    • 1995
  • Calcium hydroxide has been used not only as pulp capping and pulpotomy agents in the operative dentistry, but dressing and temporary filling materials in root canal treatment. Calcium hydroxide was known to stimulate odontoblast to produce new reparative dentin and to eliminate microorganims effectively in the infected root canals. The purpose of this study was to evaluate the effect of calcium hydroxide solution on cultured L929 cells and its antibacterial effect on several streptococci. Calcium hydroxide solution (0.121g/100ml) was added to L929 cells and cell viability was measured using 3-(4,5-dimethylthiazol-2-yl) -2,5-dimethyltetrazolium bromide (MTT) and neutral red (NR) dye. Calcium hydroxide solution (20, 40, 60, 80, 100 and $150{\mu}l$) was added to L929 cells in 96-well microplates for 1, 4 and 24 hours respectively. Cell viability was gradually decreased when the volume and exposure time of calcium hydroxide solution were increased. When $150{\mu}l$ of calcium hydroxide was applied to L929 cells for 24 hours, there was more than fifty percent reduction of cell viability. Calcium hydroxide solution (20g/100ml) showed antibacterial effect against S. uberis, S. intermedius and S. mitis after thirty-second exposure. But 0.121g/100ml concentration of cacium hydroxide solution exhibited no antibacterial effect on six streptococci after one-hour exposure.

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A Study on the Cytotoxic Effect of Heavy metals (Cd, Ni, Zn) on Cultured Mouse Fibroblast L929 Cell line (생쥐 배양섬유 모세포주 L929에 미치는 중금속(Cd, Ni, Zn)류의 세포독성에 관한 연구)

  • 이종빈;나명석;황영진;위성욱;최진희;김선희;유춘만;김재민
    • Journal of Environmental Health Sciences
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    • v.23 no.2
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    • pp.98-105
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    • 1997
  • The study on the cytotoxicity of heavy metals was carried out to evaluate the cytotoxic effect of those on mouse L929 fibroblast cell in 96-well microtiter plates. The cytotoxicity was assayed by the neutral red, tetrazolium MTT, total protein, micronuclei test. The cytotoxicity of the heavy metals by neutral red and tetrazolium MTT was showed in order, cadmium > zinc > nickel for the cationic metals tested. The effect of metal-metal interaction on the cytotoxicity showed a marked reduction of cadmium toxicity by zinc, to a lesser degree, by nickel. The amount of total protein in treated group added heavy metals was less than that of the control and treated cadmium alone was less than those of combination with nickel or zinc. At midpoint cytotoxicity values of heavy metals, the frequency of micronuclei on the cell treated heavy metals was more than that of control and treated cadmium alone was more than those of combination with nickel or zinc. From those results, it could be suggested that the heavy metals decreased the viability of mouse fibroblast L929 cells in a concentration-dependent manner and have cytogenic toxic effects, but mixed group decreased the cytotoxic and cytogenic toxicity on L929 cells.

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CYTOTOXICITY OF DENTIN BONDING SYSTEM ON L929 CELLS (상아질 결합제의 L929 세포 독성에 관한 연구)

  • Im, Mi-Kyung;Ji, Jung-Ho;Kim, Sang-Seop
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.209-219
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    • 1997
  • The research of the dentin bonding system was mainly on the chemistry and bonding strength. And in vitro assessement of biocompatibility of dentin bonding system was not completely developed. The purpose of this study was to evaluate the cytotoxic effect of several dentin primers. Scotchbond Multi-Purpose (3M Dental Products. USA). Gluma (BayerDental. Germany). All-Bond (Bisco. USA). ProBond (CaulkDensply, USA) and VeridonFil (Dongyang Nylon. Korea) were included. Cytotoxicity was tested using MTT cell viability test. 0.5 ul. 1 ul. 2 ul and 10 ul of each primer were added to the 96 well plate of incubated L929 cell lines. After 30-minute. 1. 4. 24 and 72-hour exposures. absorbance of L929 cells was observed with ELISA reader. All data were analyzed using t-test. All primers showed cytotoxicity on L929 cells under every conditions used in this study. Absorbance of L929 cells was decreased by time. Scotch bond group exhibited the lowest absorbance value in all exposure time and value.

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THE EFFECTS OF CELL WALL PROTEINS OF STREPTOCOCCUS SPP. ON DNA SYNTHESIS OF L929 CELLS AND THEIR SDS-PAGE PATTERNS (연쇄 구균의 세포벽 단백질이 L929 세포의 DNA합성에 미치는 영향 및 SDS-PAGE 양상에 관한 연구)

  • Lee, Se-Jong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.71-95
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    • 1995
  • Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

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THE EFFECT OF CELL WALL PROTEINS OF STREPTOCOCCUS SPECIES ON MICROSTRUCTURAL CHANGES OF L929 CELLS (연쇄구균의 세포벽 단백질이 L929 세포의 미세구조 변화에 미치는 영향에 관한 연구)

  • Oh, Sae-Hong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.549-576
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    • 1995
  • Bacteria are one of the most important causative agents of the pulpal and periapical diseases. Streptococci are one of the most frequently isolated facultative anarerobic bacteria in the infected root canals. Bacterial cell wall components have a direct effect in the pathogenesis of the pulpal and periapical infections. Hyaluronidase produced by bacteria has been implicated in dissemination of the diseases. The purpose of this study was to evaluate the effect of cell wall extract of streptococci on the L929 cells using inverted microscope and the transmission electron microscopy (TEM). Hyaluronidase production of streptococcal strains were investigated to determine the correlation between the severity of cell damage and the activity of enzymes. Bacterial cell wall extracts of S. sanguis, S. mitis and S. uberis isolated from infected root canals and ATCC type strains of S. mutans (ATCC 10449) and E. faecalis (ATCC 19433) were prepared by sonication and confirmed with SDS-PAGE. Silver stain of SDS-PAGE of sonic extract was efficient at $100{\mu}g$/ml concentration of cell wall protein, while Coomasie blue stain was efficient at $100{\mu}g$/ml concentration. Inverted microscope showed that sonic extract-treated L929 cells were round and detached from the substratum while others lost their fibroblastic shapes. Transmission electron microscopic examination revealed that streptococcal extracts induced death of L929 cells. Sonic extracts of streptococci had variable effect on microstructure of L929 cells. significant chromatin condensation was observed in the nucleus of the cells. Disappearance of cell surface microvilli and nuclear fragments with dense chromatin were observed. The cell nucleus had an irregular shape and numerous large vacuoles were seen in the cytoplasm and some breaks of the cell membrane could be seen. Cell organelles were in various stages of destruction and cristae of mitochondria were disoriented or disappeared. Eighteen strains of streptococci did not produce hyaluronidase.

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Development of Solid-Gelatin Microcarrier for Large Scale Production of Anchorage-Dependent Animal Gell Lines (부착성 동물세포의 대량배양을 위한 젤라틴 미립담체의 개발)

  • 박정극
    • KSBB Journal
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    • v.4 no.1
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    • pp.18-20
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    • 1989
  • Solid gelatin microcarrier with the size distribution between $100{\mu}{\textrm}{m}$ and $400{\mu}{\textrm}{m}$ was prepared for the attachment and growth experiment for anchorage-dependent animal cell lines, i.e., L 929 and BHK 21. The growth and the maximum cell densities on this gelatin based and polyacrylamide (PAA) microcarriers were compared with those on the commercial dextran based Cytodex 3 microcarrier. Both cell lines showed good comparable attachment and growth patterns on the above three microcarriers. The mouse fibroblast, L 929 showed about the same maximum cell density on PAA, gelatin and Cytodex 3 MC'S BUT BHK 21, the baby hamster kidney cell line, showed the best result on Cytodex 3, which was about $4\times10^6$ cells/ml with the microcarrier concentration of 10 g/1.

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Comparison of TNF-Mediated Glucose Catabolism between the TNF-Sensitive and -Resistant Cell Lines

  • Kim, Yeon-Hyang;Park, Bok-Ryun;Cheong, Hee-Sun;Kwon, Oh-Hwan;Kim, Dae-Que;Kim, Soung-Soo
    • BMB Reports
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    • v.32 no.2
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    • pp.140-146
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    • 1999
  • When murine fibrosarcoma L929 cells, a TNF-sensitive cell line, were treated with recombinant human tumor necrosis factor-$\alpha$ (rhTNF-$\alpha$), the activities of glycolytic regulatory enzymes and lactate dehydrogenase increased up to 100-150% compared to the control L929 cells after TNF treatment. By using various metabolic inhibitors and activators, it was found that cAMP-dependent protein kinase is responsible for the increase of activities of the glycolytic enzymes. The activities of glycolytic regulatory enzymes and lactate dehydrogenase of TNF-resistant A549 cells, a human lung carcinoma cell line, did not increase significantly compared to TNF-sensitive L929 cells upon TNF treatment. In contrast, the pyruvate carboxylase activities of A549 cells, but not L929 cells, increased up to 30~40% after TNF treatment. The data suggest that pyruvate carboxylase activity may contribute to the compensation of energy loss mediated by TNF treatment in TNF-resistant A549 cells.

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Effects of Radioprotectors on DNA Repair Capacity of Tumor Cells

  • Kim, Choon-Mi;Kim, Mi-Kyung
    • Archives of Pharmacal Research
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    • v.16 no.4
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    • pp.259-264
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    • 1993
  • Three cell lines, CHO, L929 and B16 which are non-tumorigenic and cancer cells, respectively, were first tested for their survival in the presence of radioprotective ginseng protein fraction(GPF0. The influence of three radioprotectors-CPF, cysteamine, and 1-Methyl-2-bis[(2-methylthio)vinyl] quinolinium iodide (MVQI) on DNA repair capacity of UV damaged cells survival test, the GPF showed higher cytotoxicity in L929 and B16 than in CHO cells. However, the degree of cell killing was also investigated by measuring $^3H$-thymidine incorporation of PUVA treated cells. In cell survival test, the GPF showed higher cytotoxicity in L929 and B16 than in CHO cells. However, the degree of cell killing was not high enough to consider it as an antitumorigenic agent. Variable results were obtained in the effects on DNA repair capacity depending on the protectors and cell lines used. In pretreatment, the presence of GPF and MVOI brought about a sinificant increase in the capacity in both CHO and B16 cells. However, in L929, the enhancing effect was not shown. In all three cell lines, cysteamine showed lower repair capacity than control, suggesting the primary damage reduction in stronger enhancing effects in L929 and B16 cells, while it was weaker in CHO cells. Here also cystemine hsowed a very little or no increase in the capacity in all three cell lines. These results demonstrate that GPF has mild cytotoxicity in tumorignic cells and that GPF and MVQI enhance DNA repair capacity of UV damaged cells, whether they are tumorigenic or not. On the other hand, cysteamine shows only damage reduction effect. Celles of different genetic origin seem to give different responses to the modifier and different modifiers may possibly work by different mechanisms.

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