ELF磁场对3T3细胞生长的影响及其机理的探讨

用细胞分析成像光盘记录系统测量了3T3细胞在某些ELF磁场和温度条件下生长周期的变化.实验结果表明,只有某些频率的ELF磁场才对细胞生长产生影响,磁场对细胞生长的影响还与培养细胞的生化环境有关.温度和ELF磁场都能影响细胞的生长.但二者的机理是不一样的,当撤除ELF磁场后,细胞在短时间内(2天以上)继续保持着ELF场对其生长的影响.而细胞生长周期能在短时间内(2天以内)随着温度的变化而变化.温度引起的细胞生长的变化可能与细胞内的各种生长因子、生物离子的活性有关.ELF磁场引起的细胞生长的变化可能与ELF磁场对细胞膜的影响有关,与细胞内细胞生长必不可少的生物离子(如Ca~(2+))的浓度有关.     

极低频率（ELF）磁场对细胞生长周期分布谱的影响及其机理的计算机模拟分析

测定在各种温度条件下和在ＥＬＦ（极低频率）磁场作用下细胞生长周期分布谱的变化。实验结果表明温度不仅能使周期分布谱的离散性发生改变,而且也能使谱的峰值产生位移,而ＥＬＦ磁场只能使周期分布谱的离散性发生变化,对谱的峰值没有显著影响。这一差异是由于温度和ＥＬＦ磁场对细胞生长产生影响的机理不同。用计算模拟不同机理对细胞生长周期分布谱的影响与我们早先提出的机理相吻合,即温度影响细胞内各种生长因子、生物离子的     

极低频率（ELF）磁场对细胞生长周期分布谱的影响及其机理的计算机模拟分析

测定在各种温度条件下和在ＥＬＦ（极低频率）磁场作用下细胞生长周期分布谱的变化。实验结果表明温度不仅能使周期分布谱的离散性发生改变,而且也能使谱的峰值产生位移,而ＥＬＦ磁场只能使周期分布谱的离散性发生变化,对谱的峰值没有显著影响。这一差异是由于温度和ＥＬＦ磁场对细胞生长产生影响的机理不同。用计算机模拟不同机理对细胞生长周期分布谱的影响与我们早先提出的机理相吻合,即温度影响细胞内各种生长因子、生物离子的活性,而ＥＬＦ磁场则可能通过对细胞膜的影响使细胞内的细胞生长必不可少的生物离子的浓度发生变化。     

Extremely low frequency (ELF) stray magnetic fields of laboratory equipment: a possible co-exposure conducting experiments on cell cultures

Abstract

Measurements of extremely low frequency (ELF) magnetic fields were conducted in the environment of commercial laboratory equipment in order to evaluate the possible co-exposure during the experimental processes on cell cultures. Three types of device were evaluated: a cell culture CO₂ incubator, a thermostatic water bath and a laboratory shaker table. These devices usually have electric motors, heating wires and electronic control systems, therefore may expose the cell cultures to undesirable ELF stray magnetic fields. Spatial distributions of magnetic field time domain signal waveform and frequency spectral analysis (FFT) were processed. Long- and short-term variation of stray magnetic field was also evaluated under normal use of investigated laboratory devices. The results show that the equipment under test may add a considerable ELF magnetic field to the ambient environmental magnetic field or to the intentional exposure to ELF, RF or other physical/chemical agents. The maximum stray magnetic fields were higher than 3?µT, 20?µT and 75?µT in the CO₂ incubator, in water bath and on the laboratory shaker table, respectively, with high variation of spatial distribution and time domain. Our investigation emphasizes possible confounding factors conducting cell culture studies related to low-level ELF-EMF exposure due to the existing stray magnetic fields in the ambient environment of laboratory equipment.     

ELF magnetic field affects proliferation of SPD8/V79 Chinese hamster cells but does not interact with intrachromosomal recombination

Extremely low-frequency (ELF) magnetic fields have previously been shown to affect conformation of chromatin, cell proliferation, and calcium metabolism. Possible mutagenic and carcinogenic effects of ELF have also been discussed and tested. In this study, intrachromosomal recombination in the hprt gene after exposure to ELF magnetic field was investigated using the SPD8 recombination assay. SPD8 cells, derived from V79 Chinese hamster cells were exposed to ELF at a specific combination of static and ELF magnetic fields, that has been proven to have effects on chromatin conformation in several cell types. The genotoxic agent camptothecin (CPT) was used either as a positive control or simultaneously with ELF. We also analysed the effect of ELF and CPT on chromatin conformation with the anomalous viscosity time dependence (AVTD) technique, cell growth kinetics, and cell survival with clonogenic assay. DNA fragmentation was analysed by pulsed field gel electrophoresis (PFGE). ELF did not induce recombination alone, neither did ELF modify the recombinogenic effect of CPT. Although, there was no effect on cell survival in response to ELF exposure, inhibition of cell growth was observed. On the other hand, ELF exposure partly counteracted the growth inhibition seen with CPT. The data suggest that ELF exposure may stimulate or inhibit cell growth depending on the state of the cells. Although, ELF did not induce recombination, a weak but statistically significant DNA fragmentation comparable with CPT-induced fragmentation was observed with PFGE 48h after exposure to ELF.     

Effects of extremely low-frequency magnetic field on growth and differentiation of human mesenchymal stem cells

Human Mesenchymal Stem Cells (hMSCs) were exposed to a developed extremely low-frequency (ELF) magnetic fields (50?Hz ,20?mT ELF) system to evaluate whether exposure to (ELF) magnetic fields affects growth, metabolism, and differentiation of hMSCs. MTT method was used to determine the growth and metabolism of hMSCs following exposure to ELF magnetic fields. Na(+)/K(+) concentration and osmolality of extracellular were measured after exposured culture. Alkaline phosphatase (ALP) assay and Calcium assay, ALP staining, and Alizarin red staining were performed to evaluate the osteogenic differentiation of hMSCs under the ELF magnetic field exposure. In these experiments, the cells were exposed to ELF for up to 23 days. The results showed that exposure to ELF magnetic field could inhibit the growth and metabolism of hMSC, but have no significant effect on differentiation of hMSCs. These results suggested that ELF magnetic field may influence the early development of hMSCs related adult cells.     

Effects of Extremely Low-Frequency Magnetic Field on Growth and Differentiation of Human Mesenchymal Stem Cells

Human Mesenchymal Stem Cells (hMSCs) were exposed to a developed extremely low-frequency (ELF) magnetic fields (50?Hz ,20?mT ELF) system to evaluate whether exposure to (ELF) magnetic fields affects growth, metabolism, and differentiation of hMSCs. MTT method was used to determine the growth and metabolism of hMSCs following exposure to ELF magnetic fields. Na⁺/K⁺ concentration and osmolality of extracelluar were measured after exposured culture. Alkaline phosphatase (ALP) assay and Calcium assay, ALP staining, and Alizarin red staining were performed to evaluate the osteogenic differentiation of hMSCs under the ELF magnetic field exposure. In these experiments, the cells were exposed to ELF for up to 23 days. The results showed that exposure to ELF magnetic field could inhibit the growth and metabolism of hMSC, but have no significant effect on differentiation of hMSCs. These results suggested that ELF magnetic field may influence the early development of hMSCs related adult cells.     

Inhibition of gap junction intercellular communication by extremely low-frequency electromagnetic fields in osteoblast-like models is dependent on cell differentiation.

Electromagnetic fields have been used to augment the healing of fractures because of its ability to increase new bone formation. The mechanism of how electromagnetic fields can promote new bone formation is unknown, although the interaction of electromagnetic fields with components of the plasma membrane of cells has been hypothesized to occur in bone cells. Gap junctions occur among bone forming cells, the osteoblasts, and have been hypothesized to play a role in new bone formation. Thus it was investigated whether extremely low-frequency (ELF) magnetic fields alter gap junction intercellular communication in the pre-osteoblastic model, MC3T3-E1, and the well-differentiated osteoblastic model, ROS 17/2.8. ELF magnetic field exposure systems were designed to be used for an inverted microscope stage and for a tissue culture incubator. Using these systems, it was found that magnetic fields over a frequency range from 30 to 120 Hz and field intensities up to 12.5 G dose dependently decreased gap junction intercellular communication in MC3T3-E1 cells during their proliferative phase of development. The total amount of connexin 43 protein and the distribution of connexin 43 gap junction protein between cytoplasmic and plasma membrane pools were unaltered by treatment with ELF magnetic fields. Cytosolic calcium ([Ca(2+)](i)) which can inhibit gap junction communication, was not altered by magnetic field exposure. Identical exposure conditions did not affect gap junction communication in the ROS 17/2.8 cell line and when MC3T3-E1 cells were more differentiated. Thus ELF magnetic fields may affect only less differentiated or pre-osteoblasts and not fully differentiated osteoblasts. Consequently, electromagnetic fields may aid in the repair of bone by effects exerted only on osteoprogenitor or pre-osteoblasts.     

Electromagnetic gating in ion channels.

There have been many attempts to develop a theoretical explanation of the phenomena of electromagnetic field interactions with biological systems. None of the reported efforts have been entirely successful in accounting for the observed experimental results, in particular with respect to the reports of interactions between extremely low frequency (ELF) magnetic fields and biological systems at ion cyclotron resonance frequencies. The approach used in this paper starts with the Lorentz force equation, but use is made of cylindrical co-ordinates and cylindrical boundary conditions in an attempt to more closely model the walls of an ion channel. The equations of motion of an ion that result from this approach suggest that the inside shape of the channel plus the ELF magnetic fields at specific frequencies and amplitudes could act as a gate to control the movement of the ion across the cell membrane.     

Toxicity and SOS response to ELF magnetic field and nalidixic acid in E. coli cells

Extremely low frequency (ELF) magnetic fields have previously been shown to affect conformation of chromatin and cell proliferation. Possible genotoxic and carcinogenic effects of ELF have also been discussed and tested. In this study, we analyzed the effect of ELF on chromatin conformation in E. coli GE499 cells by the anomalous viscosity time dependence (AVTD) technique. Possible genotoxic ELF effects at the specific combination of static and ELF magnetic fields, that has been proven to have effects on chromatin conformation, were investigated by clonogenic assay, cell growth kinetics, and analysis of SOS-response using inducible recA-lacZ fusion and the β-galactosidase assay. Genotoxic agent nalidixic acid (NAL) was used as positive control and in combination with ELF. Nalidixic acid at 3-30μg/ml decreased the AVTD peaks and induced cytotoxic effect. In contrast to NAL, ELF increased AVTD, stimulated cell growth, and increased cloning efficiency. These effects depended on frequency within the frequency range of 7-11Hz. While NAL induced SOS response, ELF exposure did not induce the recA-lacZ fusion. Exposure to ELF did not modify the genotoxic effects of NAL either. All together, the data show that ELF, under specific conditions of exposure, acted as nontoxic but cell growth stimulating agent.     

Applied AC and DC magnetic fields cause alterations in the mitotic cycle of early sea urchin embryos

This study demonstrates that exposure to 60 Hz magnetic fields (3.4–8.8 mT) and magnetic fields over the range DC-600 kHz (2.5–6.5 mT) can alter the early embryonic development of sea urchin embryos by inducing alterations in the timing of the cell cycle. Batches of fertilized eggs were exposed to the fields produced by a coil system. Samples of the continuous cultures were taken and scored for cell division. The times of both the first and second cell divisions were advanced by ELF AC fields and by static fields. The magnitude of the 60 Hz effect appears proportional to the field strength over the range tested. The relationship to field frequency was nonlinear and complex. For certain frequencies above the ELF range, the exposure resulted in a delay of the onset of mitosis. The advance of mitosis was also dependent on the duration of exposure and on the timing of exposure relative to fertilization. © 1995 Wiley-Liss, Inc.     

Extremely low frequency magnetic fields can either increase or decrease analgaesia in the land snail depending on field and light conditions

Results of prior investigations with opioid peptide mediated antinociception or analgaesia have suggested that these extremely low frequency (ELF) magnetic field effects are described by a resonance mechanism rather than mechanisms based on either induced currents or magnetite. Here we show that ELF magnetic fields (141-414 microT peak) can, in a manner consistent with the predictions of Lednev's parametric resonance model (PRM) for the calcium ion, either (i) reduce, (ii) have no effect on, or (iii) increase endogenous opioid mediated analgaesia in the land snail, Cepaea nemoralis. When the magnetic fields were set to parameters for the predictions of the PRM for the potassium ion, opioid-peptide mediated analgaesia increased and there was evidence of antagonism by the K(+) channel blocker, glibenclamide. Furthermore, these effects were dependent on the presence of light; the effects were absent in the absence of light. These observed increases and decreases in opioid analgaesia are largely consistent with the predictions of Lednev's PRM.     

Electric fields and surface charges induced by ELF magnetic fields

A method is described for evaluating electric fields induced by ELF magnetic fields into electrically inhomogeneous, low-conductivity (less than 5 S/m) structures. It is applied to cylinders and spheres, and numerical results are given for electrical properties that are representative of some tissues, or of cells embedded either in saline solution or a tissue matrix. Surface currents on spherical cell boundaries are estimated and compared with thermal noise due to ion motion.     

Cellular effects of electromagnetic fields

Studies at the cellular level are needed to reveal the cellular and molecular biological mechanisms underlying the biological effects and possible health implications of non-ionising radiation, such as extremely low frequency (ELF) magnetic fields (MFs) and radiofrequency (RF) fields. Our research group has studied the effects of 50 Hz ELF MFs (caused by power lines and electric devices) and 872 MHz or 900 MHz RFs (emitted by mobile phones and their base stations) on cellular ornithine decarboxylase activity, cell cycle kinetics, cell proliferation, and necrotic or apoptotic cell death. For RFs, pulse-modulated (217 Hz modulation frequency corresponding a global system for mobile communication-type signal) or continuous wave (unmodulated) signals were used. To expose the cell cultures to MFs or RFs, specially developed exposure systems were used, where levels of electromagnetic field exposure and the conditions of cell culture could be precisely controlled. A coexposure approach was used in many studies, i.e. the cell cultures were exposed to other stressors in addition to MFs or RFs. Ultraviolet radiation, serum deprivation, or fresh medium addition, were used as co-exposures. The results presented in this short review show that the effects of mere MFs or RF on cell culture models are quite minor, but that various co-exposure approaches warrant additional study.     

Frequency-dependent effects of ELF magnetic field on chromatin conformation in Escherichia coli cells and human lymphocytes.

The effects of magnetic fields of extremely low frequency (ELF, 21 microT r.m.s.) on cells of different Escherichia coli K12 strains and human lymphocytes were studied by the method of anomalous viscosity time dependence (AVTD). Within the frequency range of 6-24 Hz, two resonance-type frequency windows with maximal effects at 9 Hz and 16 Hz were observed in response of GE499 strain. Only one frequency window with maximum effect at 8.5 Hz was found for GE500 cells. These data along with previously obtained for two other E. coli strains, AB1157 and EMG2, indicate that frequency windows are dependent on genotype of cells exposed to ELF. Resonance-type effects of ELF were also observed in human lymphocytes in frequency windows around 8 and 58 Hz. These ELF effects differed significantly between studied donors, but were well reproducible in independent experiments with lymphocytes from the same donors. The frequency windows in response of E. coli strains and human lymphocytes to ELF significantly overlapped suggesting that the same targets may be involved in this response. We compared the frequency windows with predictions based on the ion cyclotron resonance (ICR) model and the magnetic parametric resonance model. These models predicted effects of ELF magnetic fields at the 'cyclotron' frequencies of some ions of biological relevance. According to the ICR model, ELF effects should be also observed at harmonics of cyclotron frequencies and, contrary, parametric resonance model predicted effects at subharmonics. While we observed coincidence of each experimental resonance frequency with predictions of one of these two models, all experimentally defined effective frequency windows were in good agreement with relatively narrow frequency ranges of both harmonics and subharmonics for natural isotopes of Na, K, Ca, Mg, and Zn ions. The experimental data support idea that both harmonics and subharmonics of several biologically important ions are involved in frequency-dependent ELF effects in cells of different types.     

Cell Sensitivity to Magnetic Fields

Attempts to establish extremely low-frequency (ELF) threshold sensitivity limits in biological systems are presently based on estimates of thermal noise in the cell membrane. The Weaver-Astumian (Science 247:459–462, 1990) threshold (8 × 10^?3 V/m) should in principle also apply to electric fields produced by Faraday induction. However, the 60-Hz magnetic field required to induce an electric field of 8 × 10^?3 V/m is improbably large and at variance with the experimental facts, implying either that Faraday induction is not the mode of weak ELF magnetic field biointeractions or that such interactions have nothing to do with the cell membrane, which constitutes only 1 % of the cell volume. We explore the possibility that magnetic field interactions are connected to the periodic changes in free calcium concentration associated with the cellular Ca²⁺ oscillator (CaO). Estimates of the free energy associated with the CaO reveal cyclic voltage changes of the order of 20 mV, suggesting that already existing electric fields within the cytoplasm may be capable of interacting with externally applied magnetic fields. We further hypothesize that CaO frequencies can be reinforced or driven into narrower passbands by weak external ELF signals acting on elements in the Ca²⁺ signaling pathway, e.g., via the calmodulin molecule.     

Combined action of static and alternating magnetic fields on ion motion in a macromolecule: Theoretical aspects

This is an attempt to solve the energetic problem of the primary detection of weak parallel static (DC) and alternating (AC) extremely low frequency (ELF) magnetic fields. We studied the equations of motion for an ion situated inside a macromolecule under the influence of these fields. The main concern is with the magnetic field influence on thermal motion of the ion in the macromolecule. The resonance effects are revealed at discrete frequencies of the ion thermal oscillations determined by the DC field magnitude and the AC field frequency. These phenomena result from the Larmor precession of the ion thermal motion. When the DC field or, to a greater extent, the combined DC and AC fields with the specific frequencies are turned on or cut off, changes occur in the energy of the ion thermal motion. If, inside the macromolecule, the ion is sufficiently protected against immediate impacts of particles of the medium surrounding the macromolecule, these changes may be enough to trigger alteration in the quantum state of the macromolecule. Bioelectromagnetics 19:279–292, 1998. © 1998 Wiley-Liss, Inc.     

Children's exposure to magnetic fields produced by U.S. television sets used for viewing programs and playing video games

Two epidemiologic studies have reported increased risk of childhood leukemia associated with the length of time children watched television (TV) programs or played video games connected to TV sets. To evaluate magnetic field exposures resulting from these activities, the static, ELF, and VLF magnetic fields produced by 72 TV sets used by children to watch TV programs and 34 TV sets used to play video games were characterized in a field study conducted in Washington DC and its Maryland suburbs. The resulting TV-specific magnetic field data were combined with information collected through questionnaires to estimate the magnetic field exposure levels associated with TV watching and video game playing. The geometric means of the ELF and VLF exposure levels so calculated were 0.0091 and 0.0016 microT, respectively, for children watching TV programs and 0.023 and 0.0038 microT, respectively, for children playing video games. Geometric means of ambient ELF and VLF levels with TV sets turned off were 0.10 and 0.0027 microT, respectively. Summed over the ELF frequency range (6-3066 Hz), the exposure levels were small compared to ambient levels. However, in restricted ELF frequency ranges (120 Hz and 606-3066 Hz) and in the VLF band, TV exposure levels were comparable to or larger than normal ambient levels. Even so, the strengths of the 120 Hz or 606-3066 Hz components of TV fields were small relative to the overall ambient levels. Consequently, our results provide little support for a linkage between childhood leukemia and exposure to the ELF magnetic fields produced by TV sets. Our results do suggest that any future research on possible health effects of magnetic fields from television sets might focus on the VLF electric and magnetic fields produced by TV sets because of their enhanced ability relative to ELF fields to induce electric currents.     

Extremely low-frequency magnetic fields of transformers and possible biological and health effects

Physiological processes in organisms can be influenced by extremely low-frequency (ELF) electromagnetic energy. Biological effect studies have great importance; as well as measurement studies since they provide information on the real exposure situations. In this study, the leakage magnetic fields around a transformer were measured in an apartment building in Küçükçekmece, Istanbul, and the measurement results were evaluated with respect to the international exposure standards. The transformer station was on the bottom floor of a three-floor building. It was found that people living and working in the building were exposed to ELF magnetic fields higher than the threshold magnetic field value of the International Agency for Research on Cancer (IARC). Many people living in this building reported health complaints such as immunological problems of their children. There were child-workers working in the textile factories located in the building. Safe distances or areas for these people should be recommended. Protective measures could be implemented to minimize these exposures. Further residential exposure studies are needed to demonstrate the exposure levels of ELF magnetic fields. Precautions should, therefore, be taken either to reduce leakage or minimize the exposed fields. Shielding techniques should be used to minimize the leakage magnetic fields in such cases.     

The role of voltage-gated Ca2+ channels in neurite growth of cultured chromaffin cells induced by extremely low frequency (ELF) magnetic field stimulation

The ion Ca²⁺ has been shown to play an important role in a wide variety of cellular functions, one of them being related to cell differentiation in which nerve growth factor (NGF) is involved. Chromaffin cells obtained from adrenals of 2- to 3-day-old rats were cultured for 7 days. During this time, these cells were subjected to the application of either NGF or extremely low frequency magnetic fields (ELF MF). Since this induced cell differentiation toward neuronal-like cells, the mechanism by which this occurred was studied. When the L-Ca²⁺ channel blocker nifedipine was applied simultaneously with ELF MF, this differentiation did not take place, but it did when an N-Ca²⁺ channel blocker was used. In contrast, none of the Ca²⁺ channel blockers prevented differentiation in the presence of NGF. In addition, Bay K-8644, an L-Ca²⁺ channel agonist, increased both the percentage of differentiated cells and neurite length in the presence of ELF MF. This effect was much weaker in the presence of NGF. [³H]-noradrenaline release was reduced by nifedipine, suggesting an important role for L-Ca²⁺ channels in neurotransmitter release. Total high voltage Ca²⁺ currents were significantly increased in ELF MF-treated cells with NGF, but these currents in ELF MF-treated cells were more sensitive to nifedipine. Amperometric analysis of catecholamine release revealed that the KCl-induced activity of cells stimulated to differentiate by ELF MF is highly sensitive to L-type Ca²⁺ channel blockers. A possible mechanism to explain the way in which the application of magnetic fields can induce differentation of chromaffin cells into neuronal-like cells is proposed.