Morphologic responses of osteoblast-like cells in monolayer culture to ELF electromagnetic fields

Osteoblast-like cells (MC 3T3-E1) were exposed for 24 h, immediately after plating, to a 60 Hz, 0.7 mT rms magnetic flux density, sufficient to induce an electric field of 0.5 mV/m rms, in order to investigate the influence of ELF field exposure on cell morphology. Using phase contrast images of the live cells, computerized image-analysis permitted rapid and objective quantification of cell length, width, area, perimeter, circularity and angular orientation. While the field-exposed cells were consistently smaller than sham treated cells, the morphologic alterations were not significantly different in the exposed cell population when cell orientation was not considered. When analyzed with respect to cell orientation, cells oriented parallel to the induced electric field (orthogonal to the applied magnetic field) demonstrated a significant decrease in cell length and an increase in roundness. These results confirm and extend previous studies on the morphologic adaptation of cells to low level ELF electromagnetic fields. The results suggest that the observed responses most likely depend on the induced electric field, with a field intensity threshold well below 1 mV/m. Further, these results provide important clues to the specific mechanism by which such low level fields may be capable of influencing cell behavior, and help to explain some of the difficulties in obtaining robust responses in in vitro EMF experiments.     

Long-term effects of 60-Hz electric vs. magnetic fields on IL-1 and IL-2 activity in sheep

This study was designed to assess the effect of exposure to long-term extremely low-frequency electric and magnetic fields (ELF-EMF) from a 500 kV transmission line on IL-1 and IL-2 activity in sheep. The primary hypothesis was that the reduction in IL-1 activity observed in our two previous short-term studies (10 months) was due to EMF exposure from this transmission line. To repeat and expand these studies and to characterize the components of EMF responsible for the previously observed reduction in IL-1 activity, the current experiment examined not only the effect of exposure to electric and magnetic fields, but also the magnetic field component alone. In the current study, IL-2 was examined to characterize the effects of EMF exposure on an indicator of T cell responses. 45 Suffolk ewe lambs were randomized into three groups of 15 animals each. One group of animals was placed in the EMF pen, located directly beneath the transmission line. A second group was placed in the shielded MF (magnetic field only) pen, also directly beneath the transmission line. The third group of animals was placed in the control pen located several hundred meters away from the transmission line. During the 27 month exposure period, blood samples were taken from all animals monthly. When the data were analyzed collectively over time, no significant differences between the groups were found for IL-1 or IL-2 activity. In previous studies ewe lambs of 8-10 weeks of age were used as the study animals and significant differences in IL-1 activity were observed after exposure of these animals to EMF at mean magnetic fields of 3.5-3.8 microT (35-38 mG) and mean electric fields of 5.2-5.8 kV/m. At the start of the current study EMF levels were reduced as compared to previous studies. One interpretation of the current data is that magnetic field strength and age of the animals may be important variables in determining whether EMF exposure will affect IL-1 activity.     

Ets1 oncogene induction by ELF-modulated 50 MHz radiofrequency electromagnetic field

We have analyzed gene expression in hemopoietic and testicular cell types after their exposure to 50 MHz radiofrequency (RF) non-ionizing radiation modulated (80%) with a 16 Hz frequency. The exposure system generates a 0.2 microT magnetic field parallel to the ground and a 60 V/m electric field orthogonal to the earth's magnetic field. Exposure conditions were selected so as to interfere with the calcium ion flow. Under these electromagnetic field (EMF) conditions, we observed an overexpression of the ets1 mRNA in Jurkat T-lymphoblastoid and Leydig TM3 cell lines. This effect was observed only in the presence of the 16 Hz modulation, corresponding to the resonance frequency for calcium ion with a DC magnetic field of 45.7 microT. We have also identified a putative candidate gene repressed after EMF exposure. The experimental model described in this paper may contribute to the understanding of the biological mechanisms involved in EMF effects.     

Evaluation of Radiofrequency and Extremely Low‐Frequency Field Levels at Children's Playground Sites in Greece From 2013 to 2018

From 2013 to 2018, in‐situ measurements of radiofrequency (RF) electromagnetic fields (EMF) and extremely low‐frequency (ELF) electric and magnetic fields in 317 existing and under‐construction children's playground facilities, in 16 municipalities all over Greece, were carried out by the Greek Atomic Energy Commission (EEAE). These measurements were conducted following legislative framework established in 2009, which requires that compliance with the established exposure limits for EMFs should be verified in playground areas. The results are presented by the value of the electric field (E) and exposure ratio (Λ) for the RF EMF, as well as the value of the electric field (E) and magnetic flux density (B) for the ELF electric and magnetic fields. Statistical analysis tools were applied on measurement data and conclusions have been made, taking into consideration: (i) environment type (urban/suburban), and (ii) vicinity to any transmitting installations. Measurement results correspond to the typical EMF background levels for each environment type. Concerning the environment type, RF EMF, and ELF electric/magnetic field measurements reveal no differentiation between urban and suburban environments. Bioelectromagnetics. 2019;40:602–605. © 2019 Bioelectromagnetics Society.     

Spindle disturbances in human-hamster hybrid (A(L) ) cells induced by the electrical component of the mobile communication frequency range signal

The production of spindle disturbances in a human–hamster hybrid (A_L) cell line by an electromagnetic field (EMF) with field strength of 90 V/m at a frequency of 900 MHz was studied in greater detail. The experimental setup presented allows investigating whether either the electrical (E) and/or the magnetic (H) field component of EMF can be associated with the effectiveness of the spindle‐disturbing potential. Therefore, both field components of a transversal electromagnetic field (TEM) wave have been separated during exposure of the biological system. This procedure should give more insight on understanding the underlying mechanisms of non‐thermal effects of EMF. A statistical comparison of the proportions of the fractions of ana‐ and telophases with spindle disturbances, obtained for five different exposure conditions with respect to unexposed controls (sham condition), showed that only cells exposed to the H‐field component of the EMF were not different from the control. Therefore, the results of the present study indicate that an exposure of cells to EMF at E‐field strengths of 45 and 90 V/m, as well as to the separated E component of the EMF, induces significant spindle disturbances in ana‐ and telophases of the cell cycle. Bioelectromagnetics 32:291–301, 2011. © 2010 Wiley‐Liss, Inc.     

No effect of extremely low-frequency magnetic field observed on cell growth or initial response of cell proliferation in human cancer cell lines

An effect on the tumor promotion process, as represented by accelerated cell growth, has been indicated as one example of areas that demonstrate the possibility of biological effects of extremely-low frequency magnetic fields. We, therefore, exposed the five cell lines (HL-60, K-562, MCF-7, A-375, and H4) derived from human tumors to a magnetic field for 3 days to investigate the effects on cell growth. Prior to exposure or sham exposure, the cells were precultured for 2 days in low serum conditions. The number of growing cells was counted in a blind manner. To investigate the effect on the initial response of cell proliferation, two cell lines were synchronized in G1 phase by serum starvation and then exposed to a magnetic field for 18 h (H4 cells) or 24 h (MCF-7 cells), both with and without serum stimulation. The rate of DNA synthesis, taken as a measure of the cell proliferation, was determined by following the incorporation of [(3)H]-thymidine into the DNA. Three different magnetic field polarizations at both 50 and 60 Hz were used: linearly polarized (vertical); circularly polarized; and an elliptically polarized field. Magnetic field flux densities were set at 500, 100, 20 and 2 microT (rms) for the vertical field and at 500 microT (rms) for the rotating fields. No effect of magnetic field exposure was observed on either cell growth or the initial response of cell proliferation.     

A scheme for incorporating DC magnetic fields into epidemiological studies of EMF exposure

Experimental data on calcium-ion release in chicken brain tissue suggest that biological effects of electric and magnetic fields (EMFs) are concentrated near certain “active combinations” of DC magnetic field strength and “effective” AC magnetic field frequencies. We hypothesize that active AC/DC combinations may exist and suggest that epidemiologic data, coupled with DC magnetic field measurements, may be used to identify critical exposure conditions. An empirical model is used to calculate these multiple active combinations at any given DC magnetic field strength and to define a rating system that incorporates the proximity of AC magnetic field frequencies generated by electric power lines to the new, computed effective frequencies. Such an exposure score may be useful in investigating correlations of EMF exposure with disease incidence. For 60 Hz and 50 Hz, the highest EMF exposure scores occurred at DC field strengths of 506 mG and 422 mG, respectively. The exposure score contains a factor which may be adjusted to reflect the importance of harmonics of the AC magnetic field as well as of the fundamental frequency. Using this factor, we consider two important special cases consistent with chick brain data: 1) we consider active pairs associated with all detectable harmonics (up to 660 Hz) without regard to relative intensity of the harmonics, and 2) we use the relative intensities of the AC field frequencies to adjust their contribution to the exposure score. © 1993 Wiley-Liss. Inc.     

Effects of extremely-law-frequency electromagnetic fields on ion transport in several mammalian cells

We have investigated the effects of sinusoidal electromagnetic fields (EMF) on ion transport (Ca²⁺, Na⁺, K⁺, and H⁺) in several cell types (red blood cells, thymocytes, Ehrlich ascites tumor cells, and HL60 and U937 human leukemia cells). The effects on the uptake of radioactive tracers as well as on the cytosolic Ca²⁺ concentration ([Ca²⁺]_i), the intracellular pH (pH_i), and the transmembrane potentsial (TMP) were studied. Exposure to EMF at 50 Hz and 100–2000 μT (rms) had no significant effects on any of these parameters. Exposure to EMF of 20–1200 μT (rms) at the estimated cyclotron magnetic resonance frequencies for the respective ions had no significant effects except for a 12–32% increase of the uptake of ⁴²K within a window at 14.5–15.5 Hz and 100–200 μT (rms), which was found in U937 and Ehrlich cells but not in the other cell types. © 1994 Wiley-Liss, Inc.     

Development and evaluation of intermediate frequency magnetic field exposure system for studies of in vitro biological effects

We have developed an intermediate frequency (IF) magnetic field exposure system for in vitro studies. Since there are no previous studies on exposure to heating-frequency magnetic fields generated from an induction heating (IH) cook top, there is a strong need for such an exposure system and for biological studies of IF magnetic fields. This system mainly consists of a magnetic-field-generating coil housed inside an incubator, inside which cultured cells can be exposed to magnetic field. Two systems were prepared to allow the experiment to be conducted in a double-blind manner. The level of the generated magnetic field was set to 532 microT rms in the exposure space, 23 kHz, 80 times the value in the International Commission on Non-ionizing Radiation Protection (ICNIRP) guidelines, with a spatial field uniformity better than 3.8%. The waveforms were nearly sinusoidal. It was also confirmed that the parasitic electric field was 157 V/m rms and the induced electric field was 1.9 V/m rms. The temperature was maintained at 36.5 +/- 0.5 degrees C for 2 h. Furthermore, leaked magnetic flux density was 0.7 microT rms or lower at extremely low frequency (ELF) and IF in the stopped system when the other system was being operated, and the environmental magnetic flux density was 0.1 microT rms or lower at the center of the coils. As a result, it was confirmed that this system could be successfully used to evaluate the biological effects of exposure to IF magnetic fields.     

Myosin Light Chain Modification Depending on Magnetic Fields: II. Experimental

Recent experiments have revealed that Ca²⁺ -calmodulin dependent myosin light chain phosphorylation in a cell-free preparation exhibits unexpectedly high sensitivity to weak magnetic fields. This enzyme system is a well-studied biochemical system, which appears to depend upon ion binding. A previous article in this journal discussed the theoretical background of myosin phosphorylation and the ion-dependent interactions of EMF with soft tissues. Because of the electromagnetic field (EMF) sensitivity of this cell-free system, experiments were designed to test the effect of a pulsed radio frequency (PRF) field, pulsating magnetic fields (TEMF), gradient magnetic fields (Magnabloc), and homogeneous static magnetic fields (in Helmholtz arrangement) designed for clinical application. It is concluded that these various magnetic fields affect this cell-free enzyme system by modulating ion–protein interactions.     

Lack of chick embryotoxicity after 20 kHz, 1.1 mT magnetic field exposure

This investigation was undertaken because biological studies to evaluate the effects of intermediate frequency magnetic fields are insufficient. White Leghorn fertile eggs (60/group) were either exposed to a 20 kHz, 1.1 mT(rms) sinusoidal magnetic field or sham‐exposed during the first 2, 7, or 11 days of embryogenesis. Lower dose exposures at 0.011 and 0.11 mT(rms) for 2 days were also conducted to elucidate possible dose–response relationships. Additional eggs given all‐trans‐retinoic acid, a teratogen, were exposed to the 1.1 mT(rms) magnetic field for the same periods to investigate the modification of embryotoxicity. After exposure, embryos were examined for mortality and developmental abnormalities. Developmental stage, number of somite pairs, and other developmental endpoints were also evaluated. Experiments were triplicated and conducted in a blind fashion. No exposure‐related changes were found in any of the endpoints in intact embryos exposed to1.1 mT(rms) or to the lower doses of 0.11 and 0.011 mT(rms) magnetic fields. Retinoic acid administration produced embryotoxic responses, which were embryonic death and developmental abnormalities, in 40–60% of embryos in the sham‐exposed groups. The magnitude of these responses was not changed significantly by the magnetic field exposures. Under the present experimental conditions, exposure to 20 kHz magnetic field up to 1.1 mT(rms) was not embryotoxic in the chick and did not potentiate the embryotoxic action of retinoic acid. Bioelectromagnetics 30:573–582, 2009. © 2009 Wiley‐Liss, Inc.     

Exposure chamber for determining the biological effects of electric and magnetic fields on dairy cows

An exposure chamber was designed to study the effects of electric and magnetic fields (EMF) on oestrous cycles, hormonal profile during gestation, pineal function, quantity and quality of milk production, feed intake, and central nervous system of dairy cattle. The chamber was 15 x 10 x 3 m; and the control system was fully computerized so that the field intensities can be varied and monitored continuously, on site or remotely. During exposure to EMF, milk production, feed consumption, and health were monitored closely and blood and cerebral spinal fluid were continuously sampled. The chamber characteristics allow use of a wide range of exposure such as electric fields (0-30 kV/m) and magnetic fields (0-100 microT) at frequencies ranging from 45 to 3000 Hz.     

Effects of sinusoidal magnetic field observed on cell proliferation, ion concentration, and osmolarity in two human cancer cell lines

Low frequency magnetic fields have previously been shown to affect cell functions. In this article, the effects of 20 mT, 50 Hz sinusoidal magnetic field on cell proliferation, ion concentration, and osmolarity in two human cancer cell lines (HL-60 and SK-Hep-1) were investigated. Inhibition of cell growth was observed. On the other hand, the exposure also increased the Na+, K+ ion concentration and osmolarity in cell supernatant compared to the control group. To our knowledge, this is the first study on cancer cells where magnetic fields affect osmolarity in cell supernatant. In addition, a model of cells exposed to the oscillating magnetic field is described as well as the characteristics of ions in and out of cells. The experimental data appears to be consistent with the theoretical analysis. The results are also discussed in terms of the relationships among cell growth, ion concentration, and osmolarity. Magnetic field inhibitions of cell growth in vitro may relate to changes in cell ion concentration and osmolarity.     

Electromagnetic effects – From cell biology to medicine

In this review we compile and discuss the published plethora of cell biological effects which are ascribed to electric fields (EF), magnetic fields (MF) and electromagnetic fields (EMF). In recent years, a change in paradigm took place concerning the endogenously produced static EF of cells and tissues. Here, modern molecular biology could link the action of ion transporters and ion channels to the “electric” action of cells and tissues. Also, sensing of these mainly EF could be demonstrated in studies of cell migration and wound healing. The triggers exerted by ion concentrations and concomitant electric field gradients have been traced along signaling cascades till gene expression changes in the nucleus.Far more enigmatic is the way of action of static MF which come in most cases from outside (e.g. earth magnetic field).All systems in an organism from the molecular to the organ level are more or less in motion. Thus, in living tissue we mostly find alternating fields as well as combination of EF and MF normally in the range of extremely low-frequency EMF. Because a bewildering array of model systems and clinical devices exits in the EMF field we concentrate on cell biological findings and look for basic principles in the EF, MF and EMF action.As an outlook for future research topics, this review tries to link areas of EF, MF and EMF research to thermodynamics and quantum physics, approaches that will produce novel insights into cell biology.     

Effect of weak combined magnetic fields on the metamorphosis of the meal-worm beetle Tenebrio molitor

The effects of weak combined magnetic fields adjusted to the parametric resonance for Ca2+ and K+ and extremely weak alternating magnetic field on the metamorphosis of the meal-worm beetle Tenebrio molitor have been studied. It was shown that the exposure of pupas of insects to all above-indicated types of fields stimulates the metamorphosis. However, after the exposure to weak combined magnetic fields adjusted to the parametric resonance for Ca2+ and K+, the number of insects with anomalies increases, which is not observed by the action of the weak alternating magnetic field.     

The effect of pulsed electromagnetic fields on the physiologic behaviour of a human astrocytoma cell line

We evaluated the effects of 50 Hz pulsed electromagnetic fields (EMFs) with a peak magnetic field of 3 mT on human astrocytoma cells. Our results clearly demonstrate that, after the cells were exposed to EMFs for 24 h, the basal [Ca(2+)](i) levels increased significantly from 124+/-51 nM to 200+/-79 nM. Pretreatment of the cells with 1.2 microM substance P increased the [Ca(2+)](i) to 555+/-278 nM, while EMF exposure caused a significant drop in [Ca(2+)](i) to 327+/-146 nM. The overall effect of EMFs probably depends on the prevailing Ca(2+) conditions of the cells. After exposure, the proliferative responses of both normal and substance P-pretreated cells increased slightly from 1.03 to 1.07 and 1.04 to 1.06, respectively. U-373 MG cells spontaneously released about 10 pg/ml of interleukin-6 which was significantly increased after the addition of substance P. Moreover, immediately after EMF exposure and 24 h thereafter, the interleukin-6 levels were more elevated (about 40%) than in controls. On the whole, our data suggest that, by changing the properties of cell membranes, EMFs can influence Ca(2+) transport processes and hence Ca(2+) homeostasis. The increased levels of interleukin-6 after 24 h of EMF exposure may confirm the complex connection between Ca(2+) levels, substance P and the cytokine network.     

ELF in vitro exposure systems for inducing uniform electric and magnetic fields in cell culture media.

Many in vitro experiments on the biological effects of extremely low frequency (ELF) electromagnetic fields utilize a uniform external magnetic flux density (B) to expose biological materials. A significant number of researchers do not measure or estimate the resulting electric field strength (E) or current density (J) in the sample medium. The magnitude and spatial distribution of the induced E field are highly dependent on the sample geometry and its relative orientation with respect to the magnetic field. We have studied the E fields induced in several of the most frequently used laboratory culture dishes and flasks under various exposure conditions. Measurements and calculations of the E field distributions in the aqueous sample volume in the containers were performed, and a set of simple, quantitative tables was developed. These tables allow a biological researcher to determine, in a straightforward fashion, the magnitudes and distributions of the electric fields that are induced in the aqueous sample when it is subjected to a uniform, sinusoidal magnetic field of known strength and frequency. In addition, we present a novel exposure technique based on a standard organ culture dish containing two circular, concentric annular rings. Exposure of the organ culture dish to a uniform magnetic field induces different average electric fields in the liquid medium in the inner and outer rings. Results of experiments with this system, which were reported in a separate paper, have shown the dominant role of the magnetically induced E field in producing specific biological effects on cells, in vitro. These results emphasize the need to report data about the induced E field in ELF in-vitro studies, involving magnetic field exposures. Our data tables on E and J in standard containers provide simple means to enable determination of these parameters.     

Test facility for human exposure to AC and DC magnetic fields

For more than a decade, Midwest Research Institute (MRI) has investigated the effects of exposure to 60 Hz electric and magnetic fields (EMF) on human physiology, performance, and biochemistry. This accumulated experience, new research directions, and limited resources made it important to design more comprehensive and operationally cost-effective exposure facilities. Here we describe the new, integrated laboratory exposure test facilities recently constructed at MRI and present data on relevant characteristics of the exposure systems. Concentric coil systems were developed to generate uniform magnetic fields within the three new exposure rooms, with rapid cancellation of the field to ambient levels in the rest of the laboratory. Control systems are fully automated, computer-based, and independent. These provide the operational flexibility needed to present fields of different magnitudes, frequencies, and polarization. The local geomagnetic field can be modulated and/or canceled, and both AC and DC fields can be presented in various combinations. Capabilities for conducting double-blind experiments with true active-sham exposure conditions were implemented using bifilar windings and applying current flow in the opposite direction for each wire in a pair. The new facilities provide a comprehensive capability for laboratory-based human research on the potential effects of exposure to AC and DC magnetic fields.     

Electromagnetic field treatment of nerve crush injury in a rat model: effect of signal configuration on functional recovery

Electromagnetic fields (EMFs) have been demonstrated to enhance mammalian peripheral nerve regeneration in vitro and in vivo. Using an EMF signal shown to enhance neurite outgrowth in vitro, we tested this field in vivo using three different amplitudes. The rat sciatic nerve was crushed. Whole body exposure was performed for 4 h/day for 5 days in a 96-turn solenoid coil controlled by a signal generator and power amplifier. The induced electric field at the target tissue consisted of a bipolar rectangular pulse, having 1 and 0.3 ms durations in each polarity, respectively. Pulse repetition rate was 2 per second. By varying the current, the coils produced fields consisting of sham (no current) and peak magnetic fields of 0.03 mT, 0.3 mT, and 3 mT, corresponding to peak induced electric fields of 1, 10, and 100 microV/cm, respectively, at the tissue target. Walking function was assessed over 43 days using video recording and measurement of the 1-5 toe-spread, using an imaging program. Comparing injured to uninjured hind limbs, mean responses were evaluated using a linear mixed statistical model. There was no difference found in recovery of the toe-spread function between any EMF treatments compared to sham.     

Effects of extremely low frequency electromagnetic fields on health

This paper gives a brief review of the physical interaction and bio-effects of exposure to extremely low frequency (ELF) electromagnetic fields (EMF) along with guidelines on limits of exposure to 50/60 Hz electric and magnetic fields.