Medical/biological study (experimental study)

ELF-EMFs induced effects on cell lines: controlling ELF generation in laboratory med./bio.

By: Farina M, Mariggio MA, Pietrangelo T, Stupak JJ, Morini A, Fano G

Published in: Progr Electromagn Res B (PIER B) 2010; 24: 131-153

Aim of study (acc. to author)

The aim of the study was twofold: 1) to investigate the production of reactive oxygen species and mitochondrial membrane potential of different cell models (neuronal, glial and skeletal muscle cells) during in vitro cell exposure to extremely low frequency electromagnetic fields and 2) to describe the protocols, in particular the equipment developed to guarantee a controlled exposure of the biological models at a 50 Hz electromagnetic field.
Additionally, data of previous studies on animal models (Falone et al. 2008) and cellular models with prokaryotic cells (Cellini et al. 2008) were also summarized. Details of those studies are not given in the present extract.

Endpoint

oxidative stress (production of reactive oxygen species and mitochondrial membrane potential)

Exposure

Exposure	Parameters
Exposure 1: 50 Hz Exposure duration: continuous for 30 min (acute exposure) or 7 days (chronic exposure)	magnetic flux density: 0.1 mT minimum magnetic flux density: 1 mT maximum

Exposure 1

Main characteristics

Frequency	50 Hz
Type	magnetic field
Waveform	sinusoidal
Exposure duration	continuous for 30 min (acute exposure) or 7 days (chronic exposure)

Exposure setup

Exposure source	Helmholtz coils
Setup	Helmholtz coils with a side length of 828 mm, a separation of 451 mm and 64 turns of AWG (American Wire Gauge) 12 wire
Sham exposure	A sham exposure was conducted.
Additional info	whether the chronic exposure was also done with Helmholtz coils or instead with a solenoid is not stated clearly in the article

Parameters

Measurand	Value	Type	Method	Mass	Remarks
magnetic flux density	0.1 mT	minimum	-	-	-
magnetic flux density	1 mT	maximum	-	-	-

Reference articles

Di Loreto S et al. (2009): Fifty hertz extremely low-frequency magnetic field exposure elicits redox and trophic response in rat-cortical neurons
Cellini L et al. (2008): Bacterial response to the exposure of 50 Hz electromagnetic fields
Falone S et al. (2008): Chronic exposure to 50Hz magnetic fields causes a significant weakening of antioxidant defence systems in aged rat brain
Falone S et al. (2007): Fifty hertz extremely low-frequency electromagnetic field causes changes in redox and differentiative status in neuroblastoma cells
Simko M et al. (2004): Extremely low frequency electromagnetic fields as effectors of cellular responses in vitro: possible immune cell activation

Exposed system:

intact cell/cell culture
PC-12 neuroblastoma cells, C2C12 cells, GL15 glioblastoma cells

Methods Endpoint/measurement parameters/methodology

cell viability/cell division/proliferation: chronic exposure experiments only: cell proliferation and cell differentiation (in the presence of NGF or low serum medium; immunoreactivity to specific differentiation markers)
oxidative stress: production of reactive oxygen species (during acute exposure; confocal microscopy via dichlorofluorescein fluorescence) and mitochondrial membrane potential (via JC-1 staining and fluorescence microscopy)

Time of investigation:

during exposure
after exposure

Main outcome of study (acc. to author)

The data indicated that the cell response (ROS production and/or mitochondrial membrane potential) after acute exposure to extremely low frequency electromagnetic fields strictly depended on the cell model rather than on the applied magnetic field strength or duration of exposure. In both neuronal-like (PC12) and glial (GL15)-like cell lines a significant increase of reactive oxygen species production was detected, with a different time-course in each cell line. In GL15 cells, this probably caused the decrease of mitochondrial membrane potential also observed in C2C12 cells.
Cell proliferation or cell differentiation did not change after chronic exposure to extremely low frequency electromagnetic fields up to 1 mT.

Study character:

medical/biological study
experimental study
pilot/exploratory/preliminary study

Calcabrini C et al. (2017): Effect of extremely low-frequency electromagnetic fields on antioxidant activity in the human keratinocyte cell line NCTC 2544
Feng B et al. (2016): Exposure to a 50-Hz magnetic field induced mitochondrial permeability transition through the ROS/GSK-3beta signaling pathway
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+
Calabro E et al. (2013): 50 Hz electromagnetic field produced changes in FTIR spectroscopy associated with mitochondrial transmembrane potential reduction in neuronal-like SH-SY5Y cells
Hong MN et al. (2012): Extremely Low Frequency Magnetic Fields Do Not Elicit Oxidative Stress in MCF10A Cells
Morabito C et al. (2010): Modulation of redox status and calcium handling by extremely low frequency electromagnetic fields in C2C12 muscle cells: A real-time, single-cell approach
Morabito C et al. (2010): Effects of Acute and Chronic Low Frequency Electromagnetic Field Exposure on PC12 Cells during Neuronal Differentiation
Di Loreto S et al. (2009): Fifty hertz extremely low-frequency magnetic field exposure elicits redox and trophic response in rat-cortical neurons
Falone S et al. (2007): Fifty hertz extremely low-frequency electromagnetic field causes changes in redox and differentiative status in neuroblastoma cells
Pozzi D et al. (2007): Effect of 50 Hz magnetic field exposure on neuroblastoma morphology
Zwirska-Korczala K et al. (2005): Effect of extremely low frequency of electromagnetic fields on cell proliferation, antioxidative enzyme activities and lipid peroxidation in 3T3-L1 preadipocytes--an in vitro study
Simko M et al. (2004): Extremely low frequency electromagnetic fields as effectors of cellular responses in vitro: possible immune cell activation