ELF/LF - Electromagnetic Fields
Experimental procedures and application ELF-EMF in regenerative medicine and cancer treatment

As in other electromagnetic fields (EMF) frequency ranges, the medical and experimental application of extremely low frequency (ELF) electromagnetic fields are used to achieve a great variety of outcomes; cellular differentiation, brain activity modulation, experimental cancer treatment, and others. ...

As all exposures revised in this website, all the intensities applied in all the experiments mentioned below are of low intensity, 1mT or less (except for a very few exceptions), even cases where much less energy is experimentally used, and being of low frequency the cause of any of the effects has nothing to do with heating and nothing to do with ionizing. Moreover there are cases, for example in bacteria, that targets are highly viable in ionizing radiation up to 5000 Gy, but their division rate is on the contrary highly sensitive to non-ionizing radiation [50], with a not established mechanism that can imply calcium ion membrane flux dynamics or ion cyclotron resonances among others.

Cellular Differentiation

One of the most interesting outcomes for a future medical application, as regenerative therapy, is the promotion of cellular differentiation, in this ambit numerous positive results have been reached, showing that ELF-EMFs can promote osteogenesis, angiogenesis, or neurogenesis using cardiac stem cells, neural stem cells, and bone narrow mesenchymal cells among others.

In [1] is proposed that the differentiation of bone marrow mesenchymal stem cells (BMSCs) into neuronal phenotype is reached via reactive oxygen production that can cause the epidermal growth factor receptor (EGFR) activation via phosphorylation and clustering, which may, in turn, lead to the activation of the PI3K/Akt signaling pathway and an increase of the CREB phosphorylation. In [51] is found that the regulation of (Zn)‐metallothionein‐3 plays a role while in [2] is found that early growth response protein 1 (Egr1) is one of the key transcription factors in that ELF-EMF-induced neuronal differentiation of the bone marrow-mesenchymal stem cells.

One of the possible medical application of the neuronal differentiation promotion is to recover brain after ischemic damage, where some experiments have auspicious results. In an experiment [3] with neural progenitor cells (NPC) data shows that ELF-EMF promotes neurogenesis of ischemic NPCs and suggest that this effect may occur through the Akt pathway. On the other hand in an experimental thesis by Gao [4] in a rat model with cerebral ischemia, it's showed that the proliferation and differentiation of neural stem cells caused after exposure probably occur by the also detected up-regulation of Hes1, Hes5 and Notch1.

Apart from recover from ischemic damage the ELF-EMF stimulation can also affects olfactory memory by modulating neurogenesis in the subventricular zone (SVZ) of the lateral ventricle of mice brain [52].

Neurogenesis of hippocampal neural stem cells with upregulation of Hes1 has also found in [5], along with upregulation of Neurogenin 1 and NeuroD1 that are strongly associated with the pan-neuronal gene expression and the neuronal fate determination.

Also, in an experimental setup with embryonic neural stem cells [6] increased expression of NeuroD and Neurogenin 1 proneural genes has been found, and also that:

" ... the expression of transient receptor potential canonical 1 (TRPC1) was significantly up-regulated accompanied by increased the peak amplitude of intracellular calcium level."

Some genes are found recursively affected along different experiments and targets, some others are detected to be affected in a novel way, in an experiment with human embryonic kidney cells they are identified 24 genes whose expression changed after ELF-EMF exposure [7], and their results points toward an important role of the histone lysine methyltransferase (Mll2), that is an enzyme that in humans is encoded by the KMT2D gene.

More interestingly, in this last study [7] they found that:

" Remarkably, an EMF-free system that eliminates Earth's naturally occurring magnetic field abrogates these epigenetic changes, resulting in a failure to undergo reprogramming."

They point out that results support a model in which the environmental magnetic field promotes chromatin reorganization through the activation of Mll2, specifically, during the dynamic epigenetic changes initiated by expression of the 4 Yamanaka reprogramming factors. Anyway, it is interesting to note here that some special properties of water, as cluster formation, requires a geomagnetic natural-like EMF exposure (in particular its natural Schumann resonance frequency at 7.8Hz) to form [8], or as is argued in some experimental procedures [9] to develop its information storing properties, see sections [10,11].

Another use for regenerative medicine is the osteogenic differentiation to regenerate bone tissues. Findings in [12] suggests that the effects of electromagnetic fields on rat BMSCs’ proliferation differentiation and mineralization are time duration dependent and that the MEK/ERK signaling pathway plays important role. Meanwhile in [13] it is showed that although when the EMF exposure is combined with an osteogenic differentiation medium the stimulation is more effective, the electromagnetic field stimulation alone also motivated the expression of osteogenic genes.

BMSCs can also be differentiated in astrocytes using ELF-EMFs; in [54] it’s found that this differentiation has been induced through the activation of SIRT1 and SIRT1 downstream molecules.

Muscle fibers can also regenerated applying ELF-EMFs on myoblast cells [55].

It’s an interesting review on dependence of Stem cell fate on electromagnetic fields [14] where the electromagnetic (EM) nature of the cells is also discussed.

Last but not least, in an experimental procedure that uses very low intensity electromagnetic fields tuned to the Ca2+ ion cyclotron resonance (ICR) at 7 Hz [15] (see this text below for a section on ICR) it is show that the exposure also induced neuronal differentiation.

Cancer

There is a very interesting line of investigation by Persinger and co-workers [16,17,18,19] where they are using physiologically-patterned ELF-EMF of very low intensity to treat cancer, inhibiting cancer cell grow and dissolving them, although with some difficulties that are trying to resolve as expressed in [17]:

" Exposure to a particular pattern of weak (~3 to 5 μT) magnetic fields produced by computer-generated point durations within three-dimensions completely dissolved malignant cancer cells but not healthy cells. Biomolecular analyses and confocal microscopy indicated excessive expansion followed by contraction contributed to the “explosion” of the cell. However, after months of replicable effects, the phenomenon slowly ceased."

The importance or patterns or the spacio-temporal component of magnetic field is also underlined in [56] where:

Cellular viability as a function of magnetic field exposure was significantly different, with a statistically smaller number of cells remaining viable after exposure to ELF-EMF than the static magnetic field, which showed no difference from controls.

A great advantage is that unlike chemical therapies and ionizing radiation, the ELF-EMF diminish the growth of only malignant cells but not normal cells. In the latest investigation of Persiger et al. [19] they confirm that the effects of electromagnetic fields on melanoma cells are dependent on their spatial and temporal character, with some configurations that provoke inhibition of cell proliferation and others with no effects, and all this using same intensities and frequencies (but differently activated over time).

The capacities of applied ELF-EMF to affect cancer specifically is very probably related to the specific endogenous electromagnetic fields of cancer cells, to this issue is a complete section [20] dedicated here, where numerous facts and theories are presented. In [21] for example it is proposed that disrupted respiration of cancer cells generate incoherent EM that in turn promote DNA strand break, and in [22] it is proposed that it can be used the enhanced electromagnetism from cancer’s centrosome clusters to attract therapeutic nanoparticles, in [23] is expressed that:

" Disturbances in oxidative metabolism and coherence are a central issue in cancer development. Oxidative metabolism may be impaired by decreased pyruvate transfer to the mitochondrial matrix, either by parasitic consumption and/or mitochondrial dysfunction. This can in turn lead to disturbance in water molecules’ ordering, diminished power, and coherence of the electromagnetic field. In tumours with the Warburg (reverse Warburg) effect, mitochondrial dysfunction affects cancer cells (fibroblasts associated with cancer cells), and the electromagnetic field generated by microtubules in cancer cells has low power (high power due to transport of energy-rich metabolites from fibroblasts), disturbed coherence, and a shifted frequency spectrum according to changed power."

Some Experimental Findings

In [24] it has been found a similar down-regulatory effect of EMF on cyclic adenosine monophosphate (cAMP) as would be seen in morphine treatment so ELF-EMF of very low intensities have potential to be used also as complementary or alternative treatment to morphine, reducing both pain and enhance patient quality.

In an experimental thesis [25] were physiologically patterned low frequency electromagnetic fields are used, it's show that exposure provoke the aggregation of bacteria in solutions, with changes in the structures of water that surround them, this effect is also seen around proteins were water is irradiated with infrared light [26] and the effect is related to the existence of Exclusion Zone waters, see section [27].

In [28] an ELF-EMF of 9 Hz was shown to exert the greatest effect on aqueous solutions of the hepatitis virus DNA amplicons, with changes in their hydration sell that suggest, again, that the aqueous milieu plays a key role as a primary target for weak effects. In this sense is very interesting the possible electromagnetic mediated DNA-Water interaction that is supported in this robust theoretical work [29], you can see this [30] section for more.

In [29] it's believed that:

" ... if we are in an environment with bio-inspired electromagnetic signals generated by mimicking natural earth and body cells frequencies (ELF's), then our cells will be more energetic and active, providing greater health … This innovative bio-inspired system has been applied for the health enhancement of humans, equines and pets etc. … It has been proven that this bio-inspired system can be effectively applied to many areas such as (1) human health enhancement and illness treatment, (2) pet health enhancement, (3) equine health treatment and (4) reduction or elimination of 'jet lag'."

Returning to the detected outcomes, in [31] it has found that the exposure to a 50-Hz magnetic field induce mitochondrial permeability transition (that can lead to mitochondrial swelling and cell death through apoptosis or necrosis depending on the particular biological setting), presumably through the ROS/GSK-3β signaling pathway. Evidences in [32] results confirmed that the ELF-EMF affects not only the ROS product but also the enzymatic activity with the modulation of catalase, cytochrome P450 and inducible nitric oxid protein expression.

In [33], after exposure of human embryonic kidney cells grown in culture, it is increased both arachidonic acid (AA) and leukotriene E4 (LTE4) levels in HEK293 cells, and is concluded that 50Hz ELF-EMF inhibits T-type calcium channels (widely expressed and that play key roles in various physiological functions like neuronal burst firing, cardiac pacemaking or secretion of hormones) through AA/LTE4 signaling pathway. The effect on those channels is also found in [16] where the promoted Ca2+ influx could be blocked by inhibitors of voltage-gated T-type Ca2+ channels. The results of [34] in cultured entorhinal cortex neurons, on the contrary, has found that exposure have to influences the intracellular calcium dynamics via a calcium channel-independent mechanism.

Very low intensities of ELF-EMF can also affect cells (and the brain as we will see later) in a variety of ways, for example in [57] they detect that the immune system can be stimulated with intensities of 0.005 mt, and in [58] using the extracellular signal-regulated kinases 1/2 (ERK1/2) activation readout in various cell lines as sensitivity detector of these cell to external ELF-MF brings the outcome that all the seven cell lines tested are sensitive to ELF-MF strengths of as low as 0.0015 mT.

Effects on Brain/Neurons

Various studies have been focused in neuronal or cerebellar cells and it is found that ELF-EMFs interact readily with the central nervous system.

In [35] Increased Na+ Currents in Rat Cerebellar Granule Cells as a result of cAMP/PKA pathway modulation was detected after 50 Hz 1mT exposure, in [36] is show that ELF-MF and ischemia separately increase oxidative stress on brains, but when applied together they have capability to decrease values it.

More generally in the brain function, relative low intensity (maximum 0.3 mT) ELF-EMFs, in the frequency range used by the brain, have show to change brain's intrinsic EEG, with for example, decreased alpha band on frontal and central areas in closed-eyes state [37]. In [38] the results have led to the authors to conclude that exposure to ELF-EMF facilitates vesicle endocytosis and synaptic plasticity in a calcium-dependent manner by increasing calcium channel expression at the nerve terminal.

Exposure to 5Hz 0.1mT [39] increased the numbers of rearing, sniffing, and locomotor activity of Wistar rats, with alterations in plasma stress hormones and glucose levels bot in 1Hz and 5Hz exposure frequencies used.

In an interesting investigation [59] electroencephalographic activity of a healthy subject originally obtained from it's quantitative electroencephalograph was recorded, those records where applied by a device to another subject’s brain diagnosed with toxic encephalopathy at really low intensities, 0.001-0.007 mT, capable however to cause significant amelioration in the subject’s diagnosis:

" In this experiment a 30 year old male university student who had been diagnosed with toxic encephalopathy six years previously and who exhibited compromised concentration, focus and processing efficiency was exposed for 30 min once per week for 6 weeks to the magnetic field equivalents of another person’s normal quantitative EEG patterns that had been recorded from each of 16 sensors. The specific magnetic field equivalents from each sensor had been reapplied through each of 16 solenoids placed in the same position over the patient’s scalp. Within two sessions there was visually conspicuous normalization of the patient’s EEG, marked reduction in the d.c. transients correlated with his distraction, and increased proficiency for scholastic performance. These results strongly suggest that applying precise spatially distributed magnetic field equivalents matched for each EEG sensor through solenoids with microTesla intensities may be able to normalize aberrant electrophysiological activity and to improve cognitive deficits."

Although the intensities used in the previous experiments are low, it has been demonstrated that the brain is capable to detect even lesser intensities, in [60] experiment demonstrated that when geomagnetic activity in the earth is at slightly altered stormy condition, even a fixed "death" brain senses it in a specific manner when the provoked microvolt fluctuations causes an increases of alpha power in the right parahippocampal gyrus:

" Here we report for the first time that fixed human tissue during specific stimulation displayed significant enhancement with geomagnetic activity and the effect was prominent within the right hemisphere. In addition we demonstrate that the specific patterns of physiologically patterned magnetic fields that are most effective for producing powerful subjective experiences also elicited the greatest response from the fixed brain tissue."

One of the possible mechanism for those subtle energy detection may reside in the ion cyclotron resonance of the different bio-molecules.

Ion Cyclotron

In a somewhat extended line of research, very low intensity ELF-EMF in the order of the geomagnetic field intensity or less are used at frequencies that correspond to ion cyclotron frequencies of specific molecules, for example in case of ca2+ [40]:

" Ca2+ ions within the specific centers of Ca2+binding proteins are the primary target of the magnetic field. Bound Ca2+ is regarded as an isotropic charged oscillator, and the MF causes precession of the axis of the Ca2+ oscillator vibration. Significant changes in the character of precession, as well as in the time average value of the degree of polarization of the oscillations of Ca2+ in a plane perpendicular to MF direction, can be induced if an alternating low frequency MF with specific resonance parameters is applied ... A low frequency MF with Ca2+ ion resonance parameters (Ca2+MF) causes a change in the Ca2+ binding constant of the protein, that is, a change in the duration of Ca2+ association with the Ca2+ binding center of the molecule, by approximately one order of magnitude."

In the mentioned experimental paper using the calculated frequency of 18.5 HZ (third harmonic of the main “cyclotron” frequency) a considerable effect on the level of activity of Ca2+ dependent proteinases is achieved, which is further confirmed in [41]. Use of the Ca2+ main cyclotron frequency at 7 Hz is decided in [15] where an important change in shape and morphology with the outgrowth of neuritic-like structures together with a lower proliferation rate and metabolic activity is achieved for human pluripotent embryonal carcinoma cells.

In the review [42], in the first table, is visible the calculated ion cyclotron frequencies, those are for 0.001 mT “environmental” static MF intensity, for other intensities simply it must be multiply, for example the ICR for Ca2+ at 0.01 mT will be 7.6 Hz. In table 2 are listed various experiments with different outcomes where Ca2+ ICR was used with intensities ranging from 0.010 mT to 0.060 mT (as comparison, earth geomagnetic fields range proximately from 0.025 mT to 0.065 mT). As mentioned, It must be said that ICR experiments also involve a static magnetic field with a strength that is in the same order of magnitude, to emulate in a controlled way a geomagnetic static MF.

In another experiment where is used the hydronium (H3O+) ion cyclotron frequency [43] it is found that:

" ... under ICR stimulation water undergoes a transition to a form that is hydroxonium-like, with the subsequent emission of a transient 48.5 Hz magnetic signal, in the absence of any other measurable field. Our results indicate that hydronium resonance stimulation alters the structure of water, enhancing the concentration of EZ-water. These results are not only consistent with Del Giudice’s model of electromagnetically coherent domains, but they can also be interpreted to show that these domains exist in quantized spin states."

For more on Exclusion Zone (EZ) water you can see the section [27].

Further confirmation of this effects come in [44] where weak magnetic field (50 nT) hydronium ICR at the field combination of 7.84 Hz, 7.5 µT, markedly changes water structure, as evidenced by the finding of an altered index of refraction exactly at this combined field.

Ion cyclotron frequencies of higher harmonics are used in a recent Nature publication [45] where ICR related frequency components significantly increased bone formation activity and it slightly increased bone resorption activity indirectly on mice. The frequencies used in this experiment are based on the following premises:

" According to ICR model, the resonant frequencies of many biologically important ions, such as Na + , K + and Ca 2+ , are intermittent frequency points and fall within 1–100 Hz 23, 25 . Apart from the fundamental frequency of resonant frequencies, when the frequency of EMF is equal to higher harmonics of the cyclotron frequencies, the biological resonant effectiveness might also be attained 26, 27 . Moreover, these higher harmonics of the cyclotron frequencies of the biologically relevant ions is blow 3,000 Hz 24 . In addition, high frequency EMF is also capable of inducing osteogenic differentiation of osteoprogenitor cells 28 . Therefore, we designed four kinds of EMF with different frequency spectrum bands (1–100 Hz, 100–3,000 Hz, 3,000–50,000 Hz and 1–50,000 Hz), among which 1–100 Hz and 100–3,000 Hz are designated as ICR frequency bands."

Ion cyclotron resonance also was used to suppress atrial fibrillation [46] in an experiment that use very low intensity fields (4 orders of magnitude less than geomagnetic fields) applied over different levels of the cardiac autonomic nervous system of dogs, and the authors believe that the effect is due to some form of subtle resonance related to neurotransmitters, they calculated ICR for vasostatin-1 a critical element in suppressing the activity of the intrinsic cardiac autonomic nervous system.

A review on ICR can be found in [47].

Related Investigations

Very related and complementary are the researches that pay more attention to the possible pernicious effects of the indiscriminate artificial ELF-EMF generated in this technological and industrial era, and that are widely employed in electrical appliances and different equipment such as television sets, mobile phones, computers and microwaves. There is a section dedicated to that [48], where it can be found, as example, an interesting study that speak about radiation effects on the secondary structure of proteins [49] among others.

References:

Very related sections:

Applied Fields - Experimental
ELF/LF - Electromagnetic Fields