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Cytotoxic effects and the mechanism of three types of magnetic nanoparticles on human hepatoma BEL-7402 cells.

Kai W, Xiaojun X, Ximing P, Zhenqing H, Qiqing Z - Nanoscale Res Lett (2011)

Bottom Line: The increase in apoptosis was accompanied with the Bax over-expression, mitochondrial membrane potential decrease, and the release of cytochrome C from mitochondria into cytosol.Our results revealed that certain concentrations of the three types of MNPs affect BEL-7402 cells viability via cell arrest and inducing apoptosis, and the MNPs-induced apoptosis is mediated through the mitochondrial-dependent pathway.The influence potency of MNPs observed in all experiments would be: C-Fe > Fe3O4 > OA-Fe3O4.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Chemistry, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, PR China. qiq@xmu.edu.cn.

ABSTRACT
The evaluation of the toxicity of magnetic nanoparticles (MNPs) has attracted much attention in recent years. The current study aimed to investigate the cytotoxic effects of Fe3O4, oleic acid-coated Fe3O4 (OA-Fe3O4), and carbon-coated Fe (C-Fe) nanoparticles on human hepatoma BEL-7402 cells and the mechanisms. WST-1 assay demonstrated that the cytotoxicity of three types of MNPs was in a dose-dependent manner. G1 (Fe3O4 and OA-Fe3O4) phase and G2 (C-Fe) phase cell arrests and apoptosis induced by MNPs were detected by flow cytometry analysis. The increase in apoptosis was accompanied with the Bax over-expression, mitochondrial membrane potential decrease, and the release of cytochrome C from mitochondria into cytosol. Moreover, apoptosis was further confirmed by morphological and biochemical hallmarks, such as swollen mitochondria with lysing cristae and caspase-3 activation. Our results revealed that certain concentrations of the three types of MNPs affect BEL-7402 cells viability via cell arrest and inducing apoptosis, and the MNPs-induced apoptosis is mediated through the mitochondrial-dependent pathway. The influence potency of MNPs observed in all experiments would be: C-Fe > Fe3O4 > OA-Fe3O4.

No MeSH data available.


Related in: MedlinePlus

The viability of BEL-7402 Cells incubated with MNPs. Cells viability was determined by WST-1 assay after BEL-7402 cells were treated with MNPs (0.01, 0.05, 0.1, 0.5, 1, and 2 mg/mL) for 24 h. The percentage of viable cells was calculated as a ratio of absorbance at 490 nm of treated to control cells.
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Figure 2: The viability of BEL-7402 Cells incubated with MNPs. Cells viability was determined by WST-1 assay after BEL-7402 cells were treated with MNPs (0.01, 0.05, 0.1, 0.5, 1, and 2 mg/mL) for 24 h. The percentage of viable cells was calculated as a ratio of absorbance at 490 nm of treated to control cells.

Mentions: 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and lactate dehydrogenase assays are frequently adopted in assessing nanoparticle toxicity. These assays are used in drug studies, but can lead to aberrant results when using nanoparticles as they can sometimes interfere with the assay components or the readout [34]. Due to its convenience and great sensitivity, recently, the WST-1 assay has become a very popular cytotoxicity assay in the nanotoxicity study [22]. After 24 h exposure at varying doses of Fe3O4, OA-Fe3O4, and C-Fe MNPs, BEL-7402 cell viabilities detected by the WST-1 assay resulted in explicit dose-dependent reduction (Figure 2). The viabilities of BEL-7402 cells exposed to all three types of MNPs were above 60% at the concentration of 0.1 mg/mL and below. When the MNPs concentrations increased more than 1 mg/mL, the cell viabilities dropped to below 60%. The viabilities of cells exposed to Fe3O4 were lower than that to OA-Fe3O4, but higher than to C-Fe at all concentrations, which were correlated with the TEM observations. The cytotoxicity is thus very likely caused by particle overload to cells [35]. It is well known that the surface of BEL-7402 cells is negatively charge. The MNPs absorbed by the cells reduced with the decrease in positively charged surfaces of MNPs due to the electrostatic effects, which could affect the amount of MNPs entering the cells and further affect cytotoxicity. In our results, the surface charge of Fe3O4, OA-Fe3O4, and C-Fe were 14.4, 4.5, and 23.7 mV, respectively, which were consistent with WST-1 data trend.


Cytotoxic effects and the mechanism of three types of magnetic nanoparticles on human hepatoma BEL-7402 cells.

Kai W, Xiaojun X, Ximing P, Zhenqing H, Qiqing Z - Nanoscale Res Lett (2011)

The viability of BEL-7402 Cells incubated with MNPs. Cells viability was determined by WST-1 assay after BEL-7402 cells were treated with MNPs (0.01, 0.05, 0.1, 0.5, 1, and 2 mg/mL) for 24 h. The percentage of viable cells was calculated as a ratio of absorbance at 490 nm of treated to control cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3211994&req=5

Figure 2: The viability of BEL-7402 Cells incubated with MNPs. Cells viability was determined by WST-1 assay after BEL-7402 cells were treated with MNPs (0.01, 0.05, 0.1, 0.5, 1, and 2 mg/mL) for 24 h. The percentage of viable cells was calculated as a ratio of absorbance at 490 nm of treated to control cells.
Mentions: 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and lactate dehydrogenase assays are frequently adopted in assessing nanoparticle toxicity. These assays are used in drug studies, but can lead to aberrant results when using nanoparticles as they can sometimes interfere with the assay components or the readout [34]. Due to its convenience and great sensitivity, recently, the WST-1 assay has become a very popular cytotoxicity assay in the nanotoxicity study [22]. After 24 h exposure at varying doses of Fe3O4, OA-Fe3O4, and C-Fe MNPs, BEL-7402 cell viabilities detected by the WST-1 assay resulted in explicit dose-dependent reduction (Figure 2). The viabilities of BEL-7402 cells exposed to all three types of MNPs were above 60% at the concentration of 0.1 mg/mL and below. When the MNPs concentrations increased more than 1 mg/mL, the cell viabilities dropped to below 60%. The viabilities of cells exposed to Fe3O4 were lower than that to OA-Fe3O4, but higher than to C-Fe at all concentrations, which were correlated with the TEM observations. The cytotoxicity is thus very likely caused by particle overload to cells [35]. It is well known that the surface of BEL-7402 cells is negatively charge. The MNPs absorbed by the cells reduced with the decrease in positively charged surfaces of MNPs due to the electrostatic effects, which could affect the amount of MNPs entering the cells and further affect cytotoxicity. In our results, the surface charge of Fe3O4, OA-Fe3O4, and C-Fe were 14.4, 4.5, and 23.7 mV, respectively, which were consistent with WST-1 data trend.

Bottom Line: The increase in apoptosis was accompanied with the Bax over-expression, mitochondrial membrane potential decrease, and the release of cytochrome C from mitochondria into cytosol.Our results revealed that certain concentrations of the three types of MNPs affect BEL-7402 cells viability via cell arrest and inducing apoptosis, and the MNPs-induced apoptosis is mediated through the mitochondrial-dependent pathway.The influence potency of MNPs observed in all experiments would be: C-Fe > Fe3O4 > OA-Fe3O4.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Chemistry, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, PR China. qiq@xmu.edu.cn.

ABSTRACT
The evaluation of the toxicity of magnetic nanoparticles (MNPs) has attracted much attention in recent years. The current study aimed to investigate the cytotoxic effects of Fe3O4, oleic acid-coated Fe3O4 (OA-Fe3O4), and carbon-coated Fe (C-Fe) nanoparticles on human hepatoma BEL-7402 cells and the mechanisms. WST-1 assay demonstrated that the cytotoxicity of three types of MNPs was in a dose-dependent manner. G1 (Fe3O4 and OA-Fe3O4) phase and G2 (C-Fe) phase cell arrests and apoptosis induced by MNPs were detected by flow cytometry analysis. The increase in apoptosis was accompanied with the Bax over-expression, mitochondrial membrane potential decrease, and the release of cytochrome C from mitochondria into cytosol. Moreover, apoptosis was further confirmed by morphological and biochemical hallmarks, such as swollen mitochondria with lysing cristae and caspase-3 activation. Our results revealed that certain concentrations of the three types of MNPs affect BEL-7402 cells viability via cell arrest and inducing apoptosis, and the MNPs-induced apoptosis is mediated through the mitochondrial-dependent pathway. The influence potency of MNPs observed in all experiments would be: C-Fe > Fe3O4 > OA-Fe3O4.

No MeSH data available.


Related in: MedlinePlus