1.Cytotoxicity and its mechanism of zinc oxide nanoparticles on human leukemic monocyte lymphoma cell line U937.
Jia-hui LIU ; Sheng-tao YANG ; Hai-fang WANG ; Yuan-fang LIU
Chinese Journal of Preventive Medicine 2010;44(9):790-794
OBJECTIVETo investigate the cytotoxicity and its mechanism of ZnO nanoparticles on human leukemic monocyte lymphoma cell line U937.
METHODSFour different size ZnO (10, 30, 60, 500 nm) were carefully characterized. The survival rate and viability were measured by trypan blue assay and MTT assay for each size ZnO particles at different concentrations (12, 120, 240, 600, 1200 µmol/L). The zinc probe, Fluozin-3, was used to detect the intracellular free zinc. Transmission electron microscopy was adopted to observe the cellular ultrastructure and the uptake of ZnO.
RESULTSAll four kinds of ZnO were rod shape, with a purity of > 99.9 wt%, and they were classified as zincite phase crystal and their surface areas were in accordance with the sizes. The viability (ZnO-n10: (97 ± 19)%, (91 ± 4)%, (24 ± 4)%, (15 ± 2)%; ZnO-n30: (111 ± 4)%, (81 ± 3)%, (24 ± 2)%, (27 ± 8)%; ZnO-n60: (105 ± 11)%, (73 ± 20)%, (43 ± 11)%, (28 ± 14)%; ZnO-µm: (88 ± 16)%, (62 ± 7)%, (22 ± 4)%, (13 ± 5)%) of cells exposed to ZnO decreased with the increasing of the concentration of ZnO from 12 to 600 µmol/L (r values were 0.965, 0.979, 0.998, 0.992, and the t values were 19.8, 25.3, 76.3, 40.9, respectively, P < 0.05). The liability (ZnO-n10: (98 ± 1)%, (67 ± 2)%, (59 ± 7)%, (13 ± 13)%, (5 ± 4)%; ZnO-n30: (98 ± 1)%, (97 ± 2)%, (50 ± 3)%, (20 ± 14)%, (7 ± 2)%; ZnO-n60: (97 ± 2)%, (88 ± 5)%, (48 ± 10)%, (12 ± 5)%, (4 ± 1)%; ZnO-µm: (96 ± 1)%, (76 ± 3)%, (58 ± 3)%, (19 ± 5)%, (20 ± 10)%) of cells exposed to ZnO decreased with the increasing of the concentration of ZnO from 12 to 600 µmol/L (r valued at 0.982, 0.956, 0.972, 0.980, and the t valued at 19.3, 12.1, 15.6, 18.5, respectively, P < 0.05). The increase of the zinc concentration showed by the zinc fluorescence probe was 121 ± 11, which was similar to the fluorescence of cells treated with ZnAc(2) (132 ± 14, F = 0.6, P > 0.05) at the Zn-equivalent concentration. There was no statistic difference for the percents of high zinc content cells in total cells exposed to ZnO-n30 (87.6 ± 2.6)% and these exposed to ZnAc(2) (86.9 ± 3.2)% (F = 1.5, P > 0.05).
CONCLUSIONZnO nanoparticles are highly cytotoxic to U937 cells and the solubilization of ZnO is the main toxicological mechanism.
Cell Survival ; Humans ; Monocytes ; drug effects ; ultrastructure ; Nanoparticles ; toxicity ; U937 Cells ; Zinc Oxide ; toxicity
2.Immunity of peritoneal monocytes against Plasmodium yoelii infected erythrocytes.
Ying-hong MAO ; Jie YUAN ; Bao-feng LIU
Acta Academiae Medicinae Sinicae 2004;26(4):415-417
OBJECTIVETo test the immunity of peritoneal monocytes against Plasmodium yoelii infected red blood cells (target cells).
METHODSSaponinized Plasmodium yoelii infected red blood cells (SPRBC, Ghost erythrocyte) were used to immunize mice i.p twice. Three weeks later, the infected red blood cells were injected i.p.; 90 min later, the total peritoneal cells were isolated and washed for scanning electromicroscopy to observe the effects of the peritoneal monocyte to the target cell.
RESULTSThe peritoneal cells of the immunized mice were activated after 90 min of the challenge of target cells. The size of the cell was not even and the pili on the cell surface turned to be long and densed. Cell interconnections were found among the cells. In some peritoneal monocytes, their cell plasma were scattered (omlette-like) or with the shape as "cellular bomb". The scattered or the sheeted pili and spredding cell plasma could adhere to the target cells which were perforated densely and damaged.
CONCLUSIONThe protective adaptive immunity exists in the peritoneal monocytes of immunized mice.
Animals ; Antibodies, Protozoan ; immunology ; Erythrocyte Membrane ; parasitology ; Female ; Malaria Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Monocytes ; immunology ; ultrastructure ; Peritoneum ; cytology ; Plasmodium yoelii ; immunology ; ultrastructure
3.In vitro model of atherosclerosis using coculture of arterial wall cells and macrophage.
Youichiro WADA ; Akira SUGIYAMA ; Takahide KOHRO ; Mika KOBAYASHI ; Motohiro TAKEYA ; Makoto NAITO ; Tatsuhiko KODAMA
Yonsei Medical Journal 2000;41(6):740-755
In order to determine the precise mechanism of the interactions between different types of cells, which are common phenomena in tissues and organs, the importance of coculture techniques are becoming increasingly important. In the area of cardiology, artificial arteries have been developed, based on the understanding of physiological communication of the arterial smooth muscle cells (SMC), endothelial cells (EC), and the extracellular matrix (ECM). In the study of atherosclerosis, the modification of low-density lipoprotein (LDL), which result in the recruitment and accumulation of white blood cells, especially, monocytes/macrophages, and foam cell formation, are hypothesized. Although there are well known animal models, an in vitro model of atherogenesis with a precisely known atherogenesis mechanism has not yet been developed. In this paper, an arterial wall reconstruction model using rabbit primary cultivated aortic SMCs and ECs, was shown. In addition, human peripheral monocytes were used and the transmigration of monocytes was observed by scanning electron and laser confocal microscopy. Monocyte differentiation into macrophages was shown by immunohistochemistry and comprehensive gene expression analysis. With the modified form of LDL, the macrophages were observed to accumulate lipids with a foamy appearance and differentiate into the foam cells in the ECM between the ECs and SMCs in the area of our coculture model.
Animal
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Aorta/physiology*
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Aorta/cytology*
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Arteriosclerosis/etiology*
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Cell Differentiation/physiology
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Cell Movement
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Coculture
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Endothelium, Vascular/physiology
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Endothelium, Vascular/cytology
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Extracellular Matrix/metabolism
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Foam Cells/ultrastructure
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Foam Cells/cytology
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Macrophages/physiology*
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Macrophages/cytology
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Male
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Microscopy, Confocal
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Microscopy, Electron
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Microscopy, Electron, Scanning
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Monocytes/ultrastructure
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Monocytes/physiology
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Muscle, Smooth, Vascular/physiology
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Muscle, Smooth, Vascular/cytology
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Myosin/metabolism
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Protein Isoforms/metabolism
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Rabbits