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Prednisolone inhibits SaOS2 osteosarcoma cell proliferation by activating inducible nitric oxide synthase

World Journal of Translational Medicine 2016年第1期5卷 53-58页

作者： Alessandra Cazzaniga jeanette am maier Sara Castiglioni Dipartimento di Scienze Biomediche e Cliniche Luigi Sacco Università di Milano

AIM:To investigate the effect of prednisolone,a synthetic glucocorticoid used in inflammatory diseases,on the growth of cultured osteosarcoma ***:Two osteosarcoma cell lines with different degree of differentiation were ***2 show a rather mature phenotype,while U2 OS are negative for almost all osteoblastic *** cells were exposed to different concentrations of prednisolone(1-9 μmol/L) with or without antioxidants or the inhibitor of inducible nitric oxide synthase(i NOS) l-N6-(iminoethyl)-lysine-HCl(L-NIL).Cell growth was assessed by counting viable *** production of nitric oxide(NO) was measured in the conditioned media by the Griess *** production of reactive oxygen species was quantified using 2'-7'-dichlorofluorescein *** blot with specific antibodies against NOSs was performed on cell ***:Prednisolone inhibited SaOS2 cell growth in a dose dependent *** significant effects were observed in *** inhibition of SaOS2 growth is not due to oxidative stress,because antioxidants do not rescue cell *** high concentrations of NO inhibit bone formation,we also measured NO and found it induced in SaOS2,but not in U2 OS,exposed to prednisolone,because of the upregulation of i NOS as detected by western ***,we treated SaOS2 with prednisolone in the presence or in the absence of L-NIL.L-NIL prevented NO release induced by prednisolone at all the concentrations apart from 9 μmol/*** the same concentrations,we found that L-NIL rescued SaOS2 growth after exposure to *** U2 OS cells,prednisolone did not induce NO production nor affected cell *** together,these data indicate that a link exists between increased amounts of NO and growth inhibition in response to prednisolone in ***:Prednisolone inhibited SaOS2 proliferation by increasing the release of NO through the upregulation of i NOS,while no effect was exerted on U2OS.

关键词： Osteosarcoma cells Prednisolone Nitric oxide Inducible nitric oxide synthase Endothelial nitric oxide synthase Reactive oxygen species

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Short- and long-term effects of silver nanoparticles on human microvascular endothelial cells

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World Journal of Biological Chemistry 2014年第4期5卷 457-464页

作者： Sara Castiglioni Clelia Caspani Alessandra Cazzaniga jeanette am maier Dipartimento di Scienze Biomediche e Cliniche Luigi Sacco Università di Milano Milano 20157 Italy

AIM: To study the response to silver nanoparticles(Ag NP) of human microvascular endothelial cells, protagonists of angiogenesis. METHODS: We cultured human microvascular endothelial cells and endothelial colony-forming cells in their corresponding growth medium. Stock solutions of Ag NP were prepared in culture medium and sonicated before use. They were added at different concentrations and for different times to culture media. The toxicity of Ag NP was investigated by measuring the reduction of yellow tetrazolium salt to dark purple formazan(MTT assay) at 575 nm. After staining with trypan blue, cell proliferation was assessed by counting viable cells. The lactate dehydrogenase leakage assay was performed on culture media by following the oxidation of NADH to NAD+ and monitoring the reaction kinetically at 340 nm. Reactive oxygen species production was quantified using 2'-7'-dichlorofluorescein diacetate. The alkaline comet assay was performed after mixing the cells with low melting-point agarose. Electrophoresis was then conducted and the samples were stained with ethidium bromide and analyzed with a fluorescence ***: Ag NP are cytotoxic in a dose and time dependent fashion for HMEC. At high concentrations, Ag NP determine loss of membrane integrity as demonstrated by the increased activity of lactate dehydrogenase in the culture medium. Ag NP rapidly stimulate the formation of free radicals. However, pre-incubation with Trolox, apocynin, or N-acetyl-L-cysteine, antioxidants which have different structure and act through different mechanisms, is not sufficient to prevent cytotoxicity. Ag NP also induce DNA damage dose-dependently, as shown by comet assay. When exposed to sublethal concentrations of Ag NP for long times, the cells remain viable but are growth retarded. Interestingly, removal of Ag NP partially rescues cell growth. Also genotoxicity is reversible upon removal of Ag NP from culture medium, suggesting that no permanent modifications oc

关键词： Endothelial cells Nanoparticles Genotoxicity Cytotoxicity Angiogenesis

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Comparison of ICare and IOPen vs Goldmann applanation tonometry according to international standards 8612 in glaucoma patients

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International Journal of Ophthalmology(English edition) 2016年第11期9卷 1624-1628页

作者： Milena Pahlitzsch jeanette Brünner Johannes Gonnermann Anna-Karina B.maier Necip Torun Eckart Bertelmann Matthias KJ.Klamann Glaucoma and Retinal Degeneration Research Group UCLInstitute of Ophthalmology Department of Ophthalmology Campus Virchow ClinicCharite University MedicineAugustenburger Platz 1

AIM： To compare IOPen and ICare rebound tonometry to Goldmann applanation tonometry（GAT） according to International Standards Organization（ISO） 8612 ***： Totally 191 eyes（n =107 individuals） were included. Criteria of ISO 8612 were fulfilled： 3 clusters of IOP, measured by GAT, were formed. The GAT results were given as mean±standard ***： GAT（19.7±0.5 mm Hg） showed a significant correlation to ICare（19.8±0.5 mm Hg）（r =0.547, P 〈0.001）and IOPen（19.5 ±0.5 mm Hg）（r =0.526, P 〈0.001）.According to ISO 8612 criteria in all 3 IOP groups the number of outliers（of the 95% limits of agreement）exceeded 5% for ICare and IOPen vs GAT： No.1（n =68）29.4% and 22.1%, No.2（n =62） 35.5% and 37.1%, No.3（n =61） 26.2% and 42.6%, ***： The strict requirements of the ISO 8612 are not fulfilled in a glaucoma collective by ICare and IOPen at present. As long as the Goldmann tonometry is applicable it should be used first of all for reproducible IOP readings. ICare and IOPen tonometry should be considered as an alternative tool, if application of Goldmann tonometry is not possible.

关键词： ICare tonometry IOPen tonometry Goldmann applanation tonometry glaucoma intraocularpressure

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