Medical/biological study (experimental study)

No evidence of DNA damage by co-exposure to extremely low frequency magnetic fields and aluminum on neuroblastoma cell lines med./bio.

By: Villarini M, Gambelunghe A, Giustarini D, Ambrosini MV, Fatigoni C, Rossi R, Dominici L, Levorato S, Muzi G, Piobbico D, Mariucci G

Published in: Mutat Res Genet Toxicol Environ Mutagen 2017; 823: 11-21

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Aim of study (acc. to author)

To investigate the effects of co-exposure to a 50 Hz magnetic field and aluminium on DNA damage in neuroblastoma cell lines.

Background/further details

In a pre-test, aluminium chloride concentrations above 40 µM were found to be cytotoxic. Therefore, concentrations up to this value were used in the experiments.
Four groups were examined for every cell line, respectively: 1) control group, 2) exposure to aluminium chloride (4 or 40 µM AlCl₃), 3) 50 Hz magnetic field exposure (0.01 mT, 0.1 mT, or 1 mT) and 4) co-exposure to aluminium chloride (4 or 40 µM AlCl₃) and 50 Hz magnetic field (0.01 mT, 0.1 mT, or 1 mT).

Endpoint

genotoxicity/mutation: DNA damage

Exposure

- 50/60 Hz
- magnetic field
- also other exposures without EMF
- co-exposure

Exposure	Parameters
Exposure 1: 50 Hz Exposure duration: 1 hour (continuously)	magnetic flux density: 0.01 mT magnetic flux density: 0.1 mT magnetic flux density: 1 mT
Exposure 2: 50 Hz Exposure duration: 5 hours (intermittent, i.e. 15 min on, 15 min off)	magnetic flux density: 0.01 mT magnetic flux density: 0.1 mT magnetic flux density: 1 mT

Exposure 1

Main characteristics

Frequency	50 Hz
Type	magnetic field
Exposure duration	1 hour (continuously)

Exposure setup

Exposure source	Helmholtz coils
Chamber	cells were exposed in 6-well plates
Setup	magnetic field was generated by an apparatus consisting of a pair of parallel coils (16.5 cm diameter (external) and 12 cm distance between the coils) arranged horizontally in a Helmholtz-configuration; well plates were positioned at the center of the coil system wit 2% field uniformity; coil system was put into a standard CO₂ incubator; temperature was 37.0°C ± 0.3°C

Parameters

Measurand	Value	Type	Method	Mass	Remarks
magnetic flux density	0.01 mT	-	measured	-	-
magnetic flux density	0.1 mT	-	measured	-	-
magnetic flux density	1 mT	-	measured	-	-

Exposure 2

Main characteristics

Frequency	50 Hz
Type	magnetic field
Exposure duration	5 hours (intermittent, i.e. 15 min on, 15 min off)

Exposure setup

Exposure source	same setup as exposure 1

Parameters

Measurand	Value	Type	Method	Mass	Remarks
magnetic flux density	0.01 mT	-	measured	-	-
magnetic flux density	0.1 mT	-	measured	-	-
magnetic flux density	1 mT	-	measured	-	-

Reference articles

Moretti M et al. (2005): Effects of co-exposure to extremely low frequency (ELF) magnetic fields and benzene or benzene metabolites determined in vitro by the alkaline comet assay

Exposed system:

intact cell/cell culture
SH-SY5Y and SK-N-BE-2 (human neuroblastoma cell lines)

Methods Endpoint/measurement parameters/methodology

genotoxicity/mutation: DNA damage (comet assay)
molecular biosynthesis: cellular redox status: level of reduced glutathione (GSH) (UV-vis HPLC) and glutathione disulfide (GSSG) (labeling with the fluorescent marker monobromobimane); protein expression of heat shock protein 70 (Hsp70) (Western blot); gene expression of heat shock protein 70 (Hsp70) (real-time RT-PCR)
cell viability/cell division/proliferation: cell viability (staining with acridine orange and DAPI; automated cytometer)

Investigated system:

Time of investigation:

after exposure

Main outcome of study (acc. to author)

No significant differences in exposed or co-exposed cells were found in any of the examined parameters compared to the control group.
The authors conclude that neither short-term exposure to aluminium chloride or to a 50 Hz magnetic field alone nor a combination of both has any detrimental effect on human neuroblastoma cells.

Study character:

medical/biological study
experimental study
full/main study
evaluation partially blinded

Study funded by

Fondazione Cassa di Risparmio di Perugia, Italy
University of Perugia, Italy

Luukkonen J et al. (2017): Modification of p21 level and cell cycle distribution by 50 Hz magnetic fields in human SH-SY5Y neuroblastoma cells
Falone S et al. (2017): Power frequency magnetic field promotes a more malignant phenotype in neuroblastoma cells via redox-related mechanisms
Benassi B et al. (2016): Extremely low frequency magnetic field (ELF-MF) exposure sensitizes SH-SY5Y cells to the pro-Parkinson's disease toxin MPP+
Kesari KK et al. (2016): Induction of micronuclei and superoxide production in neuroblastoma and glioma cell lines exposed to weak 50 Hz magnetic fields
Luukkonen J et al. (2014): Induction of genomic instability, oxidative processes, and mitochondrial activity by 50Hz magnetic fields in human SH-SY5Y neuroblastoma cells
Luukkonen J et al. (2011): Pre-Exposure to 50 Hz Magnetic Fields Modifies Menadione-Induced Genotoxic Effects in Human SH-SY5Y Neuroblastoma Cells
Villarini M et al. (2006): Effects of co-exposure to extremely low frequency (50 Hz) magnetic fields and xenobiotics determined in vitro by the alkaline comet assay
Moretti M et al. (2005): Effects of co-exposure to extremely low frequency (ELF) magnetic fields and benzene or benzene metabolites determined in vitro by the alkaline comet assay
Cho YH et al. (2003): The effect of extremely low frequency electromagnetic fields (ELF-EMF) on the frequency of micronuclei and sister chromatid exchange in human lymphocytes induced by benzo(a)pyrene
Suri A et al. (1996): A 3 milliTesla 60 Hz magnetic field is neither mutagenic nor co-mutagenic in the presence of menadione and MNU in a transgenic rat cell line

No evidence of DNA damage by co-exposure to extremely low frequency magnetic fields and aluminum on neuroblastoma cell lines med./bio.

Aim of study (acc. to author)

Background/further details

Endpoint

Exposure

Exposure 1

Main characteristics

Exposure setup

Parameters

Exposure 2

Main characteristics

Exposure setup

Parameters

Reference articles

Methods Endpoint/measurement parameters/methodology

Main outcome of study (acc. to author)

Study funded by

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