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Cytotoxicity of cobalt ferrite nanoparticles on breast cancer cell lines
S206
Abstracts / Toxicology Letters 211S (2012) S43–S216
able to induce intracellular ROS, which was in case of ZnO and CuO but not Ag NPs solely caused by dissolved metal ions. This suite of recombinant biosensors with variation in the sensing elements (specific towards other metals and stress signals) can be also used as a high-throughput screening tool for in vitro toxicological profiling of other types of metallic NPs. The results are informative not only to bacteria but also to other unicellular organisms not internalizing NPs. The support from SF0690063s08, ETF8561, ESF and FP7 project NanoValid (contract No 263147) is acknowledged. doi:10.1016/j.toxlet.2012.03.736
P33-27 Tetrahymena thermophila converts toxic silver ions to less toxic silver nanoparticles Katre Juganson 1 , Monika Mortimer 2 , Kaja Kasemets 2 , Anne Kahru 2 1
NICPB, Estonia, 2 Laboratory of Molecular Genetics, NICPB, Estonia
The use of silver nanoparticles (nanoAg) in various products, including textiles, has remarkably increased. In the current study the toxicity of AgNO3 (J.T. Baker) and nanoAg (Sigma–Aldrich) to Tetrahymena thermophila was studied as protozoa are key organisms in aerobic wastewater treatment and may become affected by (nano)silver. The 24-h EC50 value of AgNO3 to protozoa in MilliQ water was ∼3 mg/L showing their quite high tolerance to silver ions as most aquatic organisms, e.g., algae, fish and daphnids are affected by Ag-ions already at g/L level. Interestingly, during the incubation of protozoa with AgNO3 the colour of the incubation medium turned slightly maroon. NanoAg formation in the exudates of various organisms has been reported before. In order to verify that protozoan cell exudates may transform AgNO3 to nanoAg, protozoa were incubated in MilliQ for 24 h and the protozoan-free soluble extracellular fraction (SEF) was obtained using (ultra)centrifugation. AgNO3 (170 mg/L) was incubated in SEF and MilliQ (control) at 30 ◦ C for 24 h. In case of SEF-incubated AgNO3 (but not MilliQ) an absorbance peak characteristic to nanoAg at 430 nm was observed. DLS-analysis (Malvern) showed that the average hydrodynamic particle diameter was 190 nm. Thus, protozoan exudates seem to promote formation of nanoAg from Ag ions. As nanoAg was remarkably less toxic to protozoa (EC50 > 100 mg/L) this may be part of their adaptation mechanisms to toxic metal ions. SF0690063s08, ETF8561, ETF7686 and FP7 project NanoValid (contract No 263147) is acknowledged. doi:10.1016/j.toxlet.2012.03.737
P33-28 Genotoxicity of cobalt ferrite nanoparticles on MDA-231 cells using Comet assay Gonca Cakmak Demircigil 1 , Elif Asik 2 , Murat Kaya 3 , Tuba Nur Alp 3 , Murvet Volkan 3 , Mesude Iscan 4 1
Gazi University, Faculty of Pharmacy, Turkey, 2 Dept of Biotechnology, METU, Turkey, 3 Dept of Chemistry, METU, Turkey, 4 Dept of Biology, METU, Turkey
Breast cancer is the most common cancer of women; comprising 23% of all female cancers around the globe. Cobalt ferrite magnetic nanoparticles (MNPs) are attractive in cancer as imaging and therapeutic agents, and/or drug delivery vehicles. In this study, DNA
damage caused by cobalt ferrite (MNPs) were investigated on MDA231 breast cancer cells using Comet assay. Cobalt ferrite MNPs, CoFe2 O4 , with a narrow size distribution, were synthesized by co-precipitation method. MDA-231 cells which were exposed to varying concentrations of nanoparticles for 24 h were subjected to Comet Assay. Fifty randomly chosen cells from each slide were analyzed by fluorescence microscope equipped with image analyzer software. Tail intensity was used as a measure of DNA migration. The number-length mean size and volume weight mean size were calculated as, 12.9 nm and 14.06 nm, respectively, from the FE-SEM image. The polydispersivity index (PDI) value was 1.08. The zeta average mean size of the agglomerates at pH ∼ 7 was 127.4 nm. The size distribution was in the range of 50 and 200 nm. Saturation magnetization and coactivity values were measured as 46.16 emu/g and 146.974 Oe, respectively. As the MDA-231 cells were incubated with MNPs at concentrations of 17–136 g/ml for 24 h, there were no significant differences in tail intensities with respect to control cells. The tail intensities measured at each concentration were not significantly different than each other, and no concentration dependency was observed under the present experimental conditions. This work was financially supported by Project No: BAP-07-022011-004. doi:10.1016/j.toxlet.2012.03.738
P33-29 Cytotoxicity of cobalt ferrite nanoparticles on breast cancer cell lines Elif Asik 1 , Murat Kaya 2 , Tuba Nur Alp 2 , Murvet Volkan 2 , Mesude Iscan 3 1 METU, Turkey, 2 Dept of Chemistry, METU, Turkey, 3 Dept of Biology, METU, Turkey
Breast cancer is the most common cancer of women around the globe. Although there are many current treatment strategies, cobalt ferrite magnetic nanoparticles (MNPs) are attractive in breast cancer as theranostic agents. In this study, cytotoxicity of cobalt ferrite MNPs was investigated on MCF-7 and MDA-231 breast cancer cells. Cobalt ferrites MNPs, CoFe2 O4 , were synthesized by coprecipitation method. Their cytotoxicity was determined on MCF-7 and MDA-231 cells by using Cell Proliferation XTT-Kit of Biological Industries, and by tryphan blue staining. The zeta average mean size of the agglomerates at pH ∼ 7 was 127.4 nm. The size distribution was in the range of 50 and 200 nm. Saturation magnetization and coactivity values were measured as 46.16 emu/g and 146.974 Oe, respectively. The cell lines were treated with varying concentrations of nanoparticles for 24 and 48 h. Nanoparticles caused a concentration dependent decrease in viable cell numbers of both cell lines. As determined by XTT, IC50 values of nanoparticles on MCF-7 cells were calculated as 219 g/ml for 24 h and 289 g/ml for 48 h. IC50 values on MDA231 cells were found as 211 g/ml for 24 h and 189 g/ml for 48 h. IC50 values of cobalt ferrite nanoparticles were also calculated by trypan blue staining of MCF-7 cells as 268 and 311 g/ml for 24 h and 48 h, respectively. Similarly, IC50 values were determined as 293 and 302 g/ml for 24 h and 48 h treatment of MDA-231 cells, respectively, by tryphan blue staining. This work was financially supported by Project No: BAP-07-022011-004. doi:10.1016/j.toxlet.2012.03.739
Abstracts / Toxicology Letters 211S (2012) S43–S216
able to induce intracellular ROS, which was in case of ZnO and CuO but not Ag NPs solely caused by dissolved metal ions. This suite of recombinant biosensors with variation in the sensing elements (specific towards other metals and stress signals) can be also used as a high-throughput screening tool for in vitro toxicological profiling of other types of metallic NPs. The results are informative not only to bacteria but also to other unicellular organisms not internalizing NPs. The support from SF0690063s08, ETF8561, ESF and FP7 project NanoValid (contract No 263147) is acknowledged. doi:10.1016/j.toxlet.2012.03.736
P33-27 Tetrahymena thermophila converts toxic silver ions to less toxic silver nanoparticles Katre Juganson 1 , Monika Mortimer 2 , Kaja Kasemets 2 , Anne Kahru 2 1
NICPB, Estonia, 2 Laboratory of Molecular Genetics, NICPB, Estonia
The use of silver nanoparticles (nanoAg) in various products, including textiles, has remarkably increased. In the current study the toxicity of AgNO3 (J.T. Baker) and nanoAg (Sigma–Aldrich) to Tetrahymena thermophila was studied as protozoa are key organisms in aerobic wastewater treatment and may become affected by (nano)silver. The 24-h EC50 value of AgNO3 to protozoa in MilliQ water was ∼3 mg/L showing their quite high tolerance to silver ions as most aquatic organisms, e.g., algae, fish and daphnids are affected by Ag-ions already at g/L level. Interestingly, during the incubation of protozoa with AgNO3 the colour of the incubation medium turned slightly maroon. NanoAg formation in the exudates of various organisms has been reported before. In order to verify that protozoan cell exudates may transform AgNO3 to nanoAg, protozoa were incubated in MilliQ for 24 h and the protozoan-free soluble extracellular fraction (SEF) was obtained using (ultra)centrifugation. AgNO3 (170 mg/L) was incubated in SEF and MilliQ (control) at 30 ◦ C for 24 h. In case of SEF-incubated AgNO3 (but not MilliQ) an absorbance peak characteristic to nanoAg at 430 nm was observed. DLS-analysis (Malvern) showed that the average hydrodynamic particle diameter was 190 nm. Thus, protozoan exudates seem to promote formation of nanoAg from Ag ions. As nanoAg was remarkably less toxic to protozoa (EC50 > 100 mg/L) this may be part of their adaptation mechanisms to toxic metal ions. SF0690063s08, ETF8561, ETF7686 and FP7 project NanoValid (contract No 263147) is acknowledged. doi:10.1016/j.toxlet.2012.03.737
P33-28 Genotoxicity of cobalt ferrite nanoparticles on MDA-231 cells using Comet assay Gonca Cakmak Demircigil 1 , Elif Asik 2 , Murat Kaya 3 , Tuba Nur Alp 3 , Murvet Volkan 3 , Mesude Iscan 4 1
Gazi University, Faculty of Pharmacy, Turkey, 2 Dept of Biotechnology, METU, Turkey, 3 Dept of Chemistry, METU, Turkey, 4 Dept of Biology, METU, Turkey
Breast cancer is the most common cancer of women; comprising 23% of all female cancers around the globe. Cobalt ferrite magnetic nanoparticles (MNPs) are attractive in cancer as imaging and therapeutic agents, and/or drug delivery vehicles. In this study, DNA
damage caused by cobalt ferrite (MNPs) were investigated on MDA231 breast cancer cells using Comet assay. Cobalt ferrite MNPs, CoFe2 O4 , with a narrow size distribution, were synthesized by co-precipitation method. MDA-231 cells which were exposed to varying concentrations of nanoparticles for 24 h were subjected to Comet Assay. Fifty randomly chosen cells from each slide were analyzed by fluorescence microscope equipped with image analyzer software. Tail intensity was used as a measure of DNA migration. The number-length mean size and volume weight mean size were calculated as, 12.9 nm and 14.06 nm, respectively, from the FE-SEM image. The polydispersivity index (PDI) value was 1.08. The zeta average mean size of the agglomerates at pH ∼ 7 was 127.4 nm. The size distribution was in the range of 50 and 200 nm. Saturation magnetization and coactivity values were measured as 46.16 emu/g and 146.974 Oe, respectively. As the MDA-231 cells were incubated with MNPs at concentrations of 17–136 g/ml for 24 h, there were no significant differences in tail intensities with respect to control cells. The tail intensities measured at each concentration were not significantly different than each other, and no concentration dependency was observed under the present experimental conditions. This work was financially supported by Project No: BAP-07-022011-004. doi:10.1016/j.toxlet.2012.03.738
P33-29 Cytotoxicity of cobalt ferrite nanoparticles on breast cancer cell lines Elif Asik 1 , Murat Kaya 2 , Tuba Nur Alp 2 , Murvet Volkan 2 , Mesude Iscan 3 1 METU, Turkey, 2 Dept of Chemistry, METU, Turkey, 3 Dept of Biology, METU, Turkey
Breast cancer is the most common cancer of women around the globe. Although there are many current treatment strategies, cobalt ferrite magnetic nanoparticles (MNPs) are attractive in breast cancer as theranostic agents. In this study, cytotoxicity of cobalt ferrite MNPs was investigated on MCF-7 and MDA-231 breast cancer cells. Cobalt ferrites MNPs, CoFe2 O4 , were synthesized by coprecipitation method. Their cytotoxicity was determined on MCF-7 and MDA-231 cells by using Cell Proliferation XTT-Kit of Biological Industries, and by tryphan blue staining. The zeta average mean size of the agglomerates at pH ∼ 7 was 127.4 nm. The size distribution was in the range of 50 and 200 nm. Saturation magnetization and coactivity values were measured as 46.16 emu/g and 146.974 Oe, respectively. The cell lines were treated with varying concentrations of nanoparticles for 24 and 48 h. Nanoparticles caused a concentration dependent decrease in viable cell numbers of both cell lines. As determined by XTT, IC50 values of nanoparticles on MCF-7 cells were calculated as 219 g/ml for 24 h and 289 g/ml for 48 h. IC50 values on MDA231 cells were found as 211 g/ml for 24 h and 189 g/ml for 48 h. IC50 values of cobalt ferrite nanoparticles were also calculated by trypan blue staining of MCF-7 cells as 268 and 311 g/ml for 24 h and 48 h, respectively. Similarly, IC50 values were determined as 293 and 302 g/ml for 24 h and 48 h treatment of MDA-231 cells, respectively, by tryphan blue staining. This work was financially supported by Project No: BAP-07-022011-004. doi:10.1016/j.toxlet.2012.03.739