The effects of low-frequency environmental-strength electromagnetic fields on brain electrical activity: a critical review of the literature

Reports dealing with the stimulus-response relationship between low-level, low-frequency electromagnetic fields (EMFs) and changes in brain electrical activity permit assessment of the hypothesis that EMFs are detected by the body via the process of sensory transduction. These reports, as well as those involving effects on brain activity observed after a fixed time of exposure, are critically reviewed here. A consistent stimulus-response relationship between EMFs and changes in brain activity has been demonstrated in animal and human subjects. The effects, which consisted of onset and offset evoked potentials, were observed under conditions permitting the inference that the fields were transduced like ordinary stimuli such as light and sound. However, unlike the changes in brain activity induced by these stimuli, the changes induced by EMFs were governed by nonlinear laws. The studies involving attempts to determine whether a period of EMF exposure caused a metabolic effect reflected in pre-exposure/post-exposure differences in brain activity were generally inconclusive.     

Nonlinear response of the immune system to power-frequency magnetic fields

Studies of the effects of power-frequency electromagnetic fields (EMFs) on the immune and other body systems produced positive and negative results, and this pattern was usually interpreted to indicate the absence of real effects. However, if the biological effects of EMFs were governed by nonlinear laws, deterministic responses to fields could occur that were both real and inconsistent, thereby leading to both types of results. The hypothesis of real inconsistent effects due to EMFs was tested by exposing mice to 1 G, 60 Hz for 1-105 days and observing the effect on 20 immune parameters, using flow cytometry and functional assays. The data were evaluated by means of a novel statistical procedure that avoided averaging away oppositely directed changes in different animals, which we perceived to be the problem in some of the earlier EMF studies. The reliability of the procedure was shown using appropriate controls. In three independent experiments involving exposure for 21 or more days, the field altered lymphoid phenotype even though the changes in individual immune parameters were inconsistent. When the data were evaluated using traditional linear statistical methods, no significant difference in any immune parameter was found. We were able to mimic the results by sampling from known chaotic systems, suggesting that deterministic chaos could explain the effect of fields on the immune system. We conclude that exposure to power-frequency fields produced changes in the immune system that were both real and inconsistent.     

Nonlinear dynamical law governs magnetic field induced changes in lymphoid phenotype.

The results of many different types of animal and human studies dealing with the biological effects of exposure to low frequency electromagnetic fields (EMFs) have consistently been both positive and negative. We addressed the question of why this pattern had occurred so commonly in biological studies involving exposure to EMFs and hypothesized that it stemmed from the prevalent use of a linear model to characterize what are inherently nonlinear input-output relationships. The hypothesis was tested by analyzing biological data using a novel statistical procedure that could be adjusted to detect either nonlinear or linear effects. The reliability of the procedure was established using positive and negative controls and by comparison with the results obtained from sampling a known nonlinear system. In four independent experiments, male and female mice were exposed continuously to 0.1 or 0.5 mT, 60 Hz, for 175 days, and the effect on 20 immune parameters was measured using flow cytometry and functional assays. In each experiment, EMF exposure resulted in statistically significant changes in lymphoid phenotype when and only when the response of the animals to the fields was analyzed as if it were governed by nonlinear laws. Our results suggest that the pattern of inconsistency in the EMF bioeffects studies is an artifact resulting from an incorrect choice of the conceptual model for the relation between the field and the biological effect it causally determines.     

Anthropogenic Radio-Frequency Electromagnetic Fields Elicit Neuropathic Pain in an Amputation Model

Anecdotal and clinical reports have suggested that radio-frequency electromagnetic fields (RF EMFs) may serve as a trigger for neuropathic pain. However, these reports have been widely disregarded, as the epidemiological effects of electromagnetic fields have not been systematically proven, and are highly controversial. Here, we demonstrate that anthropogenic RF EMFs elicit post-neurotomy pain in a tibial neuroma transposition model. Behavioral assays indicate a persistent and significant pain response to RF EMFs when compared to SHAM surgery groups. Laser thermometry revealed a transient skin temperature increase during stimulation. Furthermore, immunofluorescence revealed an increased expression of temperature sensitive cation channels (TRPV4) in the neuroma bulb, suggesting that RF EMF-induced pain may be due to cytokine-mediated channel dysregulation and hypersensitization, leading to thermal allodynia. Additional behavioral assays were performed using an infrared heating lamp in place of the RF stimulus. While thermally-induced pain responses were observed, the response frequency and progression did not recapitulate the RF EMF effects. In vitro calcium imaging experiments demonstrated that our RF EMF stimulus is sufficient to directly contribute to the depolarization of dissociated sensory neurons. Furthermore, the perfusion of inflammatory cytokine TNF-α resulted in a significantly higher percentage of active sensory neurons during RF EMF stimulation. These results substantiate patient reports of RF EMF-pain, in the case of peripheral nerve injury, while confirming the public and scientific consensus that anthropogenic RF EMFs engender no adverse sensory effects in the general population.     

In vitro increase of the fluid-phase endocytosis induced by pulsed radiofrequency electromagnetic fields: importance of the electric field component

Nowadays, due to the wide use of mobile phones, the possible biological effects of electromagnetic fields (EMF) become a public health general concern. Despite intensive research, there are no widely accepted theories about the interactions between EMFs and living cells, and the experimental data are often controversial. We examined the effects of mobile phones EMF (envelope frequency of 217 Hz, carrier frequency of 900 MHz and pulse duration of 580 micros) or its pure, low-frequency pulsed electric field component on fluid-phase endocytosis. In both cases, with exposures exceeding 10 min, an increase of the fluid-phase endocytosis rate was observed ( approximately 1.5-fold), on three different cell types. This increase is an all-or-nothing type of response that is occurring for threshold values comprised between 1.3 and 2.6 W/kg for the delivered EMF powers and between 1.1 and 1.5 V/cm for the electric fields intensities depending upon the cell type. The electric component of these EMFs is shown to be responsible for the observed increase. Variations of frequency or pulse duration of the electric pulses are shown to be without effect. Thus, EMF, via their electrical component, can perturb one of the most fundamental physiological functions of the cells-endocytosis.     

In vitro increase of the fluid-phase endocytosis induced by pulsed radiofrequency electromagnetic fields: importance of the electric field component

Nowadays, due to the wide use of mobile phones, the possible biological effects of electromagnetic fields (EMF) become a public health general concern. Despite intensive research, there are no widely accepted theories about the interactions between EMFs and living cells, and the experimental data are often controversial. We examined the effects of mobile phones EMF (envelope frequency of 217 Hz, carrier frequency of 900 MHz and pulse duration of 580 μs) or its pure, low-frequency pulsed electric field component on fluid-phase endocytosis. In both cases, with exposures exceeding 10 min, an increase of the fluid-phase endocytosis rate was observed (≈1.5-fold), on three different cell types. This increase is an all-or-nothing type of response that is occurring for threshold values comprised between 1.3 and 2.6 W/kg for the delivered EMF powers and between 1.1 and 1.5 V/cm for the electric fields intensities depending upon the cell type. The electric component of these EMFs is shown to be responsible for the observed increase. Variations of frequency or pulse duration of the electric pulses are shown to be without effect. Thus, EMF, via their electrical component, can perturb one of the most fundamental physiological functions of the cells—endocytosis.     

Proteomic response of Schizosaccharomyces pombe to static and oscillating extremely low-frequency electromagnetic fields

There is considerable public concern regarding the health effects of exposure to low-frequency electromagnetic fields. In addition, the association between exposure and disease incidence or the possible biological effects of exposure are unclear. Using 2D-DIGE and MS in a blind study, we have investigated the effects of static and oscillating extremely low-frequency electromagnetic fields (ELF EMFs) on the proteomes of wild type Schizosaccharomyces pombe and a Sty1p deletion mutant which displays increased sensitivity to a variety of cellular stresses. Whilst this study identifies a number of protein isoforms that display significant differential expression across experimental conditions, there was no correlation between their patterns of expression and the ELF EMF exposure regimen. We conclude that there are no significant effects of either static or oscillating EMF on the yeast proteome at the sensitivity afforded by 2D-DIGE. We hypothesise that the proteins identified must be sensitive to subtle changes in culture and/or handling conditions, and that the identification of these proteins in other proteomic studies should be treated with some caution when the results of such studies are interpreted in a biological context.     

Exposure to 900 MHz electromagnetic field induces an unbalance between pro-apoptotic and pro-survival signals in T-lymphoblastoid leukemia CCRF-CEM cells

It has been recently established that low-frequency electromagnetic field (EMFs) exposure induces biological changes and could be associated with increased incidence of cancer, while the issue remains unresolved as to whether high-frequency EMFs can have hazardous effect on health. Epidemiological studies on association between childhood cancers, particularly leukemia and brain cancer, and exposure to low- and high-frequency EMF suggested an etiological role of EMFs in inducing adverse health effects. To investigate whether exposure to high-frequency EMFs could affect in vitro cell survival, we cultured acute T-lymphoblastoid leukemia cells (CCRF-CEM) in the presence of unmodulated 900 MHz EMF, generated by a transverse electromagnetic (TEM) cell, at various exposure times. We evaluated the effects of high-frequency EMF on cell growth rate and apoptosis induction, by cell viability (MTT) test, FACS analysis and DNA ladder, and we investigated pro-apoptotic and pro-survival signaling pathways possibly involved as a function of exposure time by Western blot analysis. At short exposure times (2-12 h), unmodulated 900 MHz EMF induced DNA breaks and early activation of both p53-dependent and -independent apoptotic pathways while longer continuous exposure (24-48 h) determined silencing of pro-apoptotic signals and activation of genes involved in both intracellular (Bcl-2) and extracellular (Ras and Akt1) pro-survival signaling. Overall our results indicate that exposure to 900 MHz continuous wave, after inducing an early self-defense response triggered by DNA damage, could confer to the survivor CCRF-CEM cells a further advantage to survive and proliferate.     

Extremely low frequency electromagnetic fields (ELF-EMFs) induce in vitro angiogenesis process in human endothelial cells

Effects of extremely low frequency (ELF) electromagnetic fields (EMFs) on activation of angiogenesis were analysed using cultured umbilical human vein endothelial cells (HUVECs). The cultures were exposed to a sinusoidal EMF to intensity of 1 mT, 50 Hz for up to 12 h. EMFs increased the degree of endothelial cell proliferation and tubule formation, coupled by an acceleration in the process of wound healing. Since this process is physiologically accompanied by a large modification in the structural organization of actin and focal adhesions, we analyzed the rearrangement of some cytoskeleton elements demonstrating a major reorganization of the fibres and of the focal adhesion complexes after EMF exposure. Finally, Western blot analysis revealed a significant increase in phosphorylation as well as the overall expression of VEGF receptor 2 (KDR/Flk-1) suggesting that EMFs may modulate in vitro some endothelial functions correlated to angiogenesis through signal transduction pathways dependent on VEGF.     

Diatom motility and low frequency electromagnetic fields--a new technique in the search for independent replication of results

The hypothesis that exposure to a certain combination of static and alternating electromagnetic fields (EMFs) results in an increase in motility of the marine diatom Amphora coffeaeformis was tested. Diatom motility in three strains of A. coffeaeformis was positively correlated with extracellular calcium ion (Ca2+) concentration. The test apparatus consisted of two pairs of Helmholtz coils supported around the stage of a microscope linked to a video recorder and monitor. This system allowed real-time in vivo recordings of diatom speed under EMF and control exposures. The EMFs were calculated at calcium resonance values, previously found to cause enhanced motility. Computerised image analysis was used to calculate the distance moved by individual diatoms in 2-min periods before, during and after EMF or sham-EMF (control) exposure. The addition of EMF caused no significant increase in diatom motility. The results are discussed in relation to the use of diatom motility to measure EMF exposure effects.     

Electromagnetic exposure of scaffold-free three-dimensional cell culture systems

In recent years, a number of in vitro studies have reported on the possible athermal effects of electromagnetic exposure on biological tissue. Typically, this kind of study is performed on monolayers of primary cells or cell lines. However, two‐dimensional cell layer systems lack physiological relevance since cells in vivo are organized in a three‐dimensional (3D) architecture. In monolayer studies, cell‐cell and cell‐ECM interactions obviously differ from live tissue and scale‐ups of experimental results to in vivo systems should be considered carefully. To overcome this problem, we used a scaffold‐free 3D cell culture system, suitable for the exploration of electrophysiological effects due to electromagnetic fields (EMF) at 900 MHz. Dissociated cardiac myocytes were reaggregated into cellular spheres by constant rotation, and non‐invasive extracellular recordings of these so‐called spheroids were performed with microelectrode arrays (MEA). In this study, 3D cell culture systems were exposed to pulsed EMFs in a stripline setup. We found that inhomogeneities in the EMF due to electrodes and conducting lines of the MEA chip had only a minor influence on the field distribution in the spheroid if the exposure parameters were chosen carefully. Bioelectromagnetics 32:351–359, 2011. © 2011 Wiley‐Liss, Inc.     

Exclusion zones close to wireless communication transmitters aiming to reduce human health risks

The electromagnetic fields (EMFs) radiated from typical cellular phone transmitter stations are calculated and compared to available guidelines considering thermal and non thermal biological effects. Exclusion zones close to the transmitter station are then estimated. These are three-dimensional figures calculated in comparison to a chosen EMF reference human exposure guideline limit. The volume inside them represents the regions in which the estimated electromagnetic field level is above the limit. Outside the exclusion zone the estimated electromagnetic field is below the limits. This procedure can be very useful in many applications, e.g., when a transmitter station is expected to be placed close to schools, hospitals, or close to areas where people live or work and where they spend much time each day. Owing to the uncertainty in science regarding the minimum non ionizing radiation level below which no adverse health effects can occur, the Precautionary Approach is suggested and it is recommended that EMF human exposure be kept to a minimum.     

Neural stimulation on human bone marrow‐derived mesenchymal stem cells by extremely low frequency electromagnetic fields

Adult stem cells are considered multipotent. Especially, human bone marrow‐derived mesenchymal stem cells (hBM‐MSCs) have the potential to differentiate into nerve type cells. Electromagnetic fields (EMFs) are widely distributed in the environment, and recently there have been many reports on the biological effects of EMFs. hBM‐MSCs are weak and sensitive pluripotent stem cells, therefore extremely low frequency‐electromagnetic fields (ELF‐EMFs) could be affect the changes of biological functions within the cells. In our experiments, ELF‐EMFs inhibited the growth of hBM‐MSCs in 12 days exposure. Their gene level was changed and expression of the neural stem cell marker like nestin was decreased but the neural cell markers like MAP2, NEUROD1, NF‐L, and Tau were induced. In immunofluorescence study, we confirmed the expression of each protein of neural cells. And also both oligodendrocyte and astrocyte related proteins like O4 and GFAP were expressed by ELF‐EMFs. We suggest that EMFs can induce neural differentiation in BM‐MSCs without any chemicals or differentiation factors. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

Therapeutic potential of electromagnetic fields for tissue engineering and wound healing

Ability of electromagnetic fields (EMF) to stimulate cell proliferation and differentiation has attracted the attention of many laboratories specialized in regenerative medicine over the past number of decades. Recent studies have shed light on bio‐effects induced by the EMF and how they might be harnessed to help control tissue regeneration and wound healing. Number of recent reports suggests that EMF has a positive impact at different stages of healing. Processes impacted by EMF include, but are not limited to, cell migration and proliferation, expression of growth factors, nitric oxide signalling, cytokine modulation, and more. These effects have been detected even during application of low frequencies (range: 30–300 kHz) and extremely low frequencies (range: 3–30 Hz). In this regard, special emphasis of this review is the applications of extremely low‐frequency EMFs due to their bio‐safety and therapeutic efficacy. The article also discusses combinatorial effect of EMF and mesenchymal stem cells for treatment of neurodegenerative diseases and bone tissue engineering. In addition, we discuss future perspectives of application of EMF for tissue engineering and use of metal nanoparticles activated by EMF for drug delivery and wound dressing.     

Electromagnetic fields act via activation of voltage‐gated calcium channels to produce beneficial or adverse effects

The direct targets of extremely low and microwave frequency range electromagnetic fields (EMFs) in producing non‐thermal effects have not been clearly established. However, studies in the literature, reviewed here, provide substantial support for such direct targets. Twenty‐three studies have shown that voltage‐gated calcium channels (VGCCs) produce these and other EMF effects, such that the L‐type or other VGCC blockers block or greatly lower diverse EMF effects. Furthermore, the voltage‐gated properties of these channels may provide biophysically plausible mechanisms for EMF biological effects. Downstream responses of such EMF exposures may be mediated through Ca²⁺/calmodulin stimulation of nitric oxide synthesis. Potentially, physiological/therapeutic responses may be largely as a result of nitric oxide‐cGMP‐protein kinase G pathway stimulation. A well‐studied example of such an apparent therapeutic response, EMF stimulation of bone growth, appears to work along this pathway. However, pathophysiological responses to EMFs may be as a result of nitric oxide‐peroxynitrite‐oxidative stress pathway of action. A single such well‐documented example, EMF induction of DNA single‐strand breaks in cells, as measured by alkaline comet assays, is reviewed here. Such single‐strand breaks are known to be produced through the action of this pathway. Data on the mechanism of EMF induction of such breaks are limited; what data are available support this proposed mechanism. Other Ca²⁺‐mediated regulatory changes, independent of nitric oxide, may also have roles. This article reviews, then, a substantially supported set of targets, VGCCs, whose stimulation produces non‐thermal EMF responses by humans/higher animals with downstream effects involving Ca²⁺/calmodulin‐dependent nitric oxide increases, which may explain therapeutic and pathophysiological effects.     

Radiofrequency electromagnetic fields do not alter the cell cycle progression of C3H 10T and U87MG cells.

The effects of exposure to radiofrequency electromagnetic fields (RF EMFs) on cell cycle progression of mouse fibroblasts C3H 10T(1/2) and human glioma U87MG cells were determined by the flow cytometric bromodeoxyuridine pulse-chase method. Cells were exposed to a frequency-modulated continuous wave at 835.62 MHz or a code division multiple access RF EMF centered on 847.74 MHz at an average specific absorption rate of 0.6 W/kg. Five cell cycle parameters, including the transit of cells through G(1), G(2) and S phase and the probability of cell division, were examined immediately after the cells were placed in the fields or after they had been kept in the fields for up to 100 h. The only significant change observed in the study was that associated with C3H 10T(1/2) cell cultures moving into plateau phase toward the later times in the long-exposure experiment. No changes in the cell cycle parameters were observed in cells exposed to either mode of RF EMFs when compared to sham-exposed cells in either of the cell lines studied during the entire experimental period. The results show that exposure to RF EMFs, at the frequencies and power tested, does not have any effect on cell progression in vitro.     

Assessing of plasma protein denaturation induced by exposure to cadmium,electromagnetic fields and their combined actions on rat

In our environment, we have numerous chances to be exposed to not only electromagnetic fields (EMFs) but also many chemicals containing mutagens. Therefore, the aim of this study was to estimate whether rat’s exposure to cadmium and/or EMFs could cause oxidative damage to molecular structure of proteins and whether and to what extent the effects of co-exposure differ from those observed under the treatment with each exposure alone. Thirty-two rats were divided into four groups. Group 1 was termed as control, group 2 was treated with cadmium (3.0?mg/Kg), group 3 was exposed to EMF (10?mT/h/day) and group 4 was treated with cadmium and exposed to EMF. Protein carbonyls (PCO) in the plasma as a marker of oxidative protein damage and total oxidant status (TOS), as well as electrical conductivity and SDS electrophoresis to estimate changes in molecular structure of protein, were determined. The exposure to Cd and/or EMF led to oxidative protein damage (increased PCO and TOS) accomplished by increased stress of electrical charges on the surface of the protein molecule (increased electrical conductivity) and changes in the molecular structure of protein. The effects were more pronounced after treatment with both Cd and EMF than at the treatment with each exposure alone. The serious damage to proteins at the co-exposure to Cd and EMF seems to be due to the interference of the EMF with the toxic activity of cadmium. This work concluded that combined exposure to Cd and EMFs might increase the risk of plasma damage via enhancing free radical generation and protein oxidation.     

Impact of electromagnetic fields on in vitro toxicity of silver and graphene nanoparticles

The correlation between shape and concentration of silver nanoparticles (AgNPs), their cytotoxicity and formation of reactive oxygen species (ROS) in the presence of electromagnetic fields (EMFs) has been investigated. In addition, the bio-effects caused by the combination of EMFs and graphene nanoparticles (GrNPs) have been also assessed. The AgNPs of three shapes (triangular, spherical and colloidal) and GrNPs were added in high concentrations to the culture of human fibroblasts and exposed to EMF of three different frequencies: 900, 2400 and 7500 MHz. The results demonstrated the dependence of the EMF-induced cytotoxicity on the shape and concentration of AgNPs. The maximal cell killing effect was observed at 900 MHz frequency for NPs of all shapes and concentrations. The highest temperature elevation was observed for GrNPs solution irradiated by EMF of 900 MHz frequency. The exposure to EMF led to significant increase of ROS formation in triangular and colloidal AgNPs solutions. However, no impact of EMF on ROS production was detected for spherical AgNPs. GrNPs demonstrated ROS-protective activity that was dependent on their concentration. Our findings indicate the feasibility to control cytotoxicity of AgNPs by means of EMFs. The effect EMF on the biological activity of AgNPs and GrNPs is reported here for the first time.     

Comparative study of cell cycle kinetics and induction of apoptosis or necrosis after exposure of human Mono Mac 6 cells to radiofrequency radiation

The possible harmful effects of radiofrequency electromagnetic fields (RF EMFs) are controversial. We have used human Mono Mac 6 cells to investigate the influence of RF EMFs in vitro on cell cycle alterations and BrdU uptake, as well as the induction of apoptosis and necrosis in human Mono Mac 6 cells, using flow cytometry after exposure to a 1,800 MHz, 2 W/kg specific absorption rate (SAR), GSM-DTX signal for 12 h. No statistically significant differences in the induction of apoptosis or necrosis, cell cycle kinetics, or BrdU uptake were detected after RF EMF exposure compared to sham or incubator controls. However, in the positive control cells treated with gliotoxin and PMA (phorbol 12 myristate-13 acetate), a significant increase in apoptotic and necrotic cells was seen. Cell cycle analysis or BrdU incorporation for 72 h showed no differences between RF EMF- or sham-exposed cells, whereas PMA treatment induced a significant accumulation of cells in G(0)/G(1)-phase and a reduction in S-phase cells. RF EMF radiation did not induce cell cycle alterations or changes in BrdU incorporation or induce apoptosis and necrosis in Mono Mac 6 cells under the exposure conditions used.