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Cytotoxicity of iron oxide nanoparticles with different coatings on human neuronal cells
Abstracts / Toxicology Letters 229S (2014) S40–S252
epigenetic effects of NPs exposure in vivo. We exposed BALB/C mice to well-characterized gold NPs (Au NPs) of 5 nm, 60 nm and 250 nm diameter; single-walled carbon nanotubes (SWCNTs) and multi-walled carbon nanotubes (MWCNTs) at two dose levels (high dose: 1 mg/ml; low dose: 100 g/ml) for 48 h. Control mice received either vehicle (0.2% saline) or air (sham-control). Mice were sacrificed after 48 h, and lung and blood cells were analyzed for epigenetic endpoints i.e., global and gene specific methylation, and DNA hydroxymethylation. NPs exposure showed alterations in global DNA hydroxymethylation in mice lung DNA while no alterations were observed for global DNA methylation. NPs exposure in mice also altered methylation level of number of genes in lung and blood tissues. The epigenetically altered genes were involved in DNA repair pathway, cell cycle control pathway, oxidative stress response pathway and immune related pathways. We are further translating in vivo findings in humans who are exposed to NPs. In conclusion our results indicated the involvement of epigenetic factors in response to NPs exposure in vivo. http://dx.doi.org/10.1016/j.toxlet.2014.06.672 P-3.160 Cytotoxicity of iron oxide nanoparticles with different coatings on human neuronal cells Carla Costa 1,2 , Gozde Kilic¸ 3,4 , Fátima Brandão 1 , Maria João Bessa 1 , Solange Costa 1,2 , Eduardo Pásaro 3 , Vanessa Valdiglesias 3 , Blanca Laffon 3 , João Paulo Teixeira 1,2,∗ 1
Department of Environmental Health, Portuguese National Institute of Health, Porto, Portugal, 2 Institute of Public Health, University of Porto, Porto, Portugal, 3 DICOMOSA Group, Department of Psychology, Area of Psychobiology, Universidade da Coru˜ na, A Coru˜ na, Spain, 4 Department of Cell and Molecular Biology, Universidade da Coru˜ na, A Coru˜ na, Spain The rapid development of nanotechnology has led to the wide application of nanoparticles (NPs) in various fields. Superparamagnetic iron oxide NPs have different biomedical applications, like as contrast agents in magnetic resonance imaging. After systemic administration, NPs are small enough to penetrate small capillaries and offer an effective approach to target certain tissues such as the brain. In this work, human neuroblastoma SHSY5Y cells were exposed to different concentrations of two iron oxide magnetite NPs (one coated with silica and another with oleic acid) (range 5–300 g/ml), prepared in complete cell culture medium and incomplete cell culture medium (serum-free) for three exposure times (3, 6, 24 h). Cytotoxicity was evaluated after exposure to NPs by means of the MTT, AB, LDH and NRU assays. Characterisation of the main physical–chemical properties of the tested NPs was also performed. Exposure to magnetite NPs coated with oleic acid for 24 h treatments was responsible for a significant increase in the percentage of LDH, both in incomplete and complete media. Silica-coated magnetite NPs prepared in incomplete medium also induced a decrease in viability of SHSY5Y cells at the highest concentrations in the 24 h treatments, assessed by MTT and NRU assays. Although results suggest that these NPs are biocompatible at low concentrations, further studies are necessary to examine possible genotoxic effects even at low concentrations. Research was funded by Xunta de Galicia (EM 2012/079) and by TD1204 MODENA COST Action. G. Kilic¸ was supported by a fellow˜ ship from the University of A Coruna. http://dx.doi.org/10.1016/j.toxlet.2014.06.673
S199
P-3.161 Evaluation of potency of antibacterial activity of some -lactum antibiotics conjugated with CDSE/ZNS quantum dots Manmohan Satnami, Sandeep Vaishanav ∗ Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India -Lactum antibiotics are broad class of antibiotics which kills bacteria by inhibiting the formation of peptidoglycan that constitutes the bacterial cell wall. The resistance that develops in bacteria for antibiotics led the scientific world to think about the future aspects for modifying the way through which antibiotics are acted on the bacteria and become lethal for them. Nowadays nanodrug delivery system for targeted delivery of a drug is the hottest area of research. In this consequence, we try to evaluate the potential of latest marketed antibiotics that e.g. Amoxiciline (I), ceftazidim(II) after being conjugated with quantum dots. The mechanism of QD-conjugated antibiotics against the bacteria has been investigated. We have synthesized CdSe/ZnS quantum dots that have been characterized through various spectroscopic techniques viz. UV–vis spectroscopy, fluorescence measurements, FTIR analysis, X-ray diffraction (XRD) study and transmission electron microscopy (TEM). The surface of quantum dots have been conjugated with antibiotics by corbodiimied coupling with the help of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) as conjugating agent between antibiotic and functionalized quantum dots. The antibacterial properties of QD-conjugated antibiotics have been determined by disc diffusion assay. The potency of QD-conjugated antibiotics have been estimated by determining their IC50 for the selected strain of gram negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi. http://dx.doi.org/10.1016/j.toxlet.2014.06.674 P-3.162 Effects of single and repeated oral administration of mesoporous silica nanoparticles in Wistar rats Virginia Tzankova, Vessela Vitcheva ∗ Faculty of Pharmacy, Medical University – Sofia, Laboratory of “Drug metabolism and drug toxicity”, Department of “Pharmacology and Toxicology”, Sofia, Bulgaria Mesoporous silica nanoparticles (MCM-41) are relatively new and promising nanomaterials. They have been widely studied as drug carriers, because of their unique mesoporous structure and greater surface area. However, there are very few studies in regard to the toxicological potential of MCM-41 in vivo. On the basis of these data, the aim of the following study was to investigate the oral toxicity of MCM-41 after single (100 mg/kg, bw) and 14 days (10 mg/kg, bw) administration in male Wistar rats. The production of thiobarbituric acid reactive substances, measured as malonedialdehide (MDA), level of reduced glutathione (GSH) and activities of the following antioxidant enzymes: glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST) were measured in liver homogenate spectrophotometrically. Hematological (Er, Ht, Leuc, Plt) and blood biochemical parameters (ASAT, ALAT, LDH, Alb, Urea) were also assessed.
epigenetic effects of NPs exposure in vivo. We exposed BALB/C mice to well-characterized gold NPs (Au NPs) of 5 nm, 60 nm and 250 nm diameter; single-walled carbon nanotubes (SWCNTs) and multi-walled carbon nanotubes (MWCNTs) at two dose levels (high dose: 1 mg/ml; low dose: 100 g/ml) for 48 h. Control mice received either vehicle (0.2% saline) or air (sham-control). Mice were sacrificed after 48 h, and lung and blood cells were analyzed for epigenetic endpoints i.e., global and gene specific methylation, and DNA hydroxymethylation. NPs exposure showed alterations in global DNA hydroxymethylation in mice lung DNA while no alterations were observed for global DNA methylation. NPs exposure in mice also altered methylation level of number of genes in lung and blood tissues. The epigenetically altered genes were involved in DNA repair pathway, cell cycle control pathway, oxidative stress response pathway and immune related pathways. We are further translating in vivo findings in humans who are exposed to NPs. In conclusion our results indicated the involvement of epigenetic factors in response to NPs exposure in vivo. http://dx.doi.org/10.1016/j.toxlet.2014.06.672 P-3.160 Cytotoxicity of iron oxide nanoparticles with different coatings on human neuronal cells Carla Costa 1,2 , Gozde Kilic¸ 3,4 , Fátima Brandão 1 , Maria João Bessa 1 , Solange Costa 1,2 , Eduardo Pásaro 3 , Vanessa Valdiglesias 3 , Blanca Laffon 3 , João Paulo Teixeira 1,2,∗ 1
Department of Environmental Health, Portuguese National Institute of Health, Porto, Portugal, 2 Institute of Public Health, University of Porto, Porto, Portugal, 3 DICOMOSA Group, Department of Psychology, Area of Psychobiology, Universidade da Coru˜ na, A Coru˜ na, Spain, 4 Department of Cell and Molecular Biology, Universidade da Coru˜ na, A Coru˜ na, Spain The rapid development of nanotechnology has led to the wide application of nanoparticles (NPs) in various fields. Superparamagnetic iron oxide NPs have different biomedical applications, like as contrast agents in magnetic resonance imaging. After systemic administration, NPs are small enough to penetrate small capillaries and offer an effective approach to target certain tissues such as the brain. In this work, human neuroblastoma SHSY5Y cells were exposed to different concentrations of two iron oxide magnetite NPs (one coated with silica and another with oleic acid) (range 5–300 g/ml), prepared in complete cell culture medium and incomplete cell culture medium (serum-free) for three exposure times (3, 6, 24 h). Cytotoxicity was evaluated after exposure to NPs by means of the MTT, AB, LDH and NRU assays. Characterisation of the main physical–chemical properties of the tested NPs was also performed. Exposure to magnetite NPs coated with oleic acid for 24 h treatments was responsible for a significant increase in the percentage of LDH, both in incomplete and complete media. Silica-coated magnetite NPs prepared in incomplete medium also induced a decrease in viability of SHSY5Y cells at the highest concentrations in the 24 h treatments, assessed by MTT and NRU assays. Although results suggest that these NPs are biocompatible at low concentrations, further studies are necessary to examine possible genotoxic effects even at low concentrations. Research was funded by Xunta de Galicia (EM 2012/079) and by TD1204 MODENA COST Action. G. Kilic¸ was supported by a fellow˜ ship from the University of A Coruna. http://dx.doi.org/10.1016/j.toxlet.2014.06.673
S199
P-3.161 Evaluation of potency of antibacterial activity of some -lactum antibiotics conjugated with CDSE/ZNS quantum dots Manmohan Satnami, Sandeep Vaishanav ∗ Pt. Ravishankar Shukla University, Raipur, Chhattisgarh, India -Lactum antibiotics are broad class of antibiotics which kills bacteria by inhibiting the formation of peptidoglycan that constitutes the bacterial cell wall. The resistance that develops in bacteria for antibiotics led the scientific world to think about the future aspects for modifying the way through which antibiotics are acted on the bacteria and become lethal for them. Nowadays nanodrug delivery system for targeted delivery of a drug is the hottest area of research. In this consequence, we try to evaluate the potential of latest marketed antibiotics that e.g. Amoxiciline (I), ceftazidim(II) after being conjugated with quantum dots. The mechanism of QD-conjugated antibiotics against the bacteria has been investigated. We have synthesized CdSe/ZnS quantum dots that have been characterized through various spectroscopic techniques viz. UV–vis spectroscopy, fluorescence measurements, FTIR analysis, X-ray diffraction (XRD) study and transmission electron microscopy (TEM). The surface of quantum dots have been conjugated with antibiotics by corbodiimied coupling with the help of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) as conjugating agent between antibiotic and functionalized quantum dots. The antibacterial properties of QD-conjugated antibiotics have been determined by disc diffusion assay. The potency of QD-conjugated antibiotics have been estimated by determining their IC50 for the selected strain of gram negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi. http://dx.doi.org/10.1016/j.toxlet.2014.06.674 P-3.162 Effects of single and repeated oral administration of mesoporous silica nanoparticles in Wistar rats Virginia Tzankova, Vessela Vitcheva ∗ Faculty of Pharmacy, Medical University – Sofia, Laboratory of “Drug metabolism and drug toxicity”, Department of “Pharmacology and Toxicology”, Sofia, Bulgaria Mesoporous silica nanoparticles (MCM-41) are relatively new and promising nanomaterials. They have been widely studied as drug carriers, because of their unique mesoporous structure and greater surface area. However, there are very few studies in regard to the toxicological potential of MCM-41 in vivo. On the basis of these data, the aim of the following study was to investigate the oral toxicity of MCM-41 after single (100 mg/kg, bw) and 14 days (10 mg/kg, bw) administration in male Wistar rats. The production of thiobarbituric acid reactive substances, measured as malonedialdehide (MDA), level of reduced glutathione (GSH) and activities of the following antioxidant enzymes: glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST) were measured in liver homogenate spectrophotometrically. Hematological (Er, Ht, Leuc, Plt) and blood biochemical parameters (ASAT, ALAT, LDH, Alb, Urea) were also assessed.