PEMF and atherosclerosis

Atherosclerosis serves as the core pathological basis of cardiovascular, cerebrovascular, and peripheral arterial diseases, posing a serious threat to human health. However, current mainstream treatments such as statin drugs and stent implantation are associated with significant side effects or limited efficacy, highlighting the urgent need for new therapeutic strategies. Pulsed electromagnetic fields (PEMFs), due to their noninvasive nature and anti-inflammatory properties, show potential in the treatment of atherosclerosis. Cheng 2025 is a mega paper that this post cannot possibly properly dissect. In some ways the methodology is the most interesting aspect of the study.

Cheng H, Zhang Q, Zhong W, Li H, Wang L, Wang S, He C, Fu C, Wei Q. Pulsed electromagnetic fields inhibit atherosclerosis by regulating pyroptosis through membrane tension-mediated mechanosensitive channels. Signal Transduct Target Ther. 2025 Nov 28;10(1):388. PMC free paper

the graphical abstract in bullet points

Click on the above link to view a picture summary of what was concluded.

• Atherosclerosis prone ApoE-/- mice, ApoE-/-NLRP3^-/- knockout mice were used. Human umbilical vein endothelial cells (HUVECs), human aortic endothelial cells (HAECs) were used as a more easily manipulated model.
• When lipids build up on the vessel walls membrane tension is increased activating the membrane tension and the mechanosensitive tension-mediated transient receptor potential vanilloid 4 (TRPV4) channels.
• The increase in Ca2+ through the TRPV4 channels causes mitochondria dysfunction and generation of reactive oxygen species. (ROS).
• ROS activate infalmmasomes leading to the cleavage of inhibitory peptides of cytokines IL-18 and IL-1β. The N-terminus of GSDMD is cleaved. GSDMD-Nt forms membrane pores in a process called pyroptosis.
  
• Human plasma samples were analyzed suggesting some sort of membrane protein difference in normal controls and those with atherosclerosis.
• PEMFs were found to effectively inhibit the activation of the NLRP3 inflammasome, reduce plaque formation, and delay the progression of atherosclerosis.

“This discovery not only reveals a novel mechanobiological pathway but also provides a solid theoretical foundation for the development of PEMF-based therapies for atherosclerosis.”

Atherosclerosis background

This section is a background on peripheral artery disease secondary to atherosclerosis compiled separately from Cheng 2025. This is an example of a large vessel that is prone to peripheral artery disease atherosclerosis. Wikipedia’s piece on thefemoral artery:

“The common femoral artery (CFA) is located between the inferior margin of the inguinal ligament, and the branching point of the deep femoral artery. Its first three or four centimeters are enclosed, with the femoral vein, in the femoral sheath.” The CFA basically lives in the upper thigh. The CFA is also prone to calcification that cannot easily be resolved with a stent. An extensive review of the literature was conducted by vascular surgeons from NYU Langone Health Long Island and Stonybrook who concluded that blasting plaques with sound waves, lithotripsy, is a way around stents that cannot be placed in the common femoral artery (Ye 2025). Ye and the other vascular surgeons claim that the gold standard is endarterectomy, cutting the plaque out. Wikipedia authors do a good job of explaining the basics of the surgery and the problems elaborated by Ye 2025. Shockwavemedical.com is one of the few sources of the actual soundwave frequencies I could find. Their waveform is about 14 microseconds in length, or about 71 kHz ultrasound.

Ye IB, Giannopoulos S, Kokkosis AA, Jasinski PT, Labropoulos N. Challenging the No-Stent Zone: Intravascular Lithotripsy for Common Femoral Artery Disease. J Clin Med. 2025 Sep 15;14(18):6492. PMC free paper

A previous post covered the use of PEMF for neovascularization when a major artery is occluded in a rodent model of peripheral artery disease.

Cheng 2025 Materials and Methods

Cheng 2025 is about the intersection of inflammasomes (no adenosine), mitochondria (no complex IV), and TRPV4 Ca²⁺ channels that are distinct from what PEMF hacks think of voltage gated Ca²⁺channels. The strength of the conclusions of a study rest on the reliability of the methods.

Lipid Deposits

Oil Red O simply stains lipids red. H&E staining detects a variety of different cellular structures. Hematoxylin stains calcium deposits red while eosin cytoplasm and the lipid parts of plaques pink. Cheng 2025 is the first PEMF paper to my knowledge demonstrate prevention of lipid build up anywhere. It is noted that they did not look for their oxidized low density lipoprotein binding to cells in their cell culture model.

Membrane tension and the cytoskeleton

Flipper basically detects changes in membrane fluidity. Think fats in the refrigerator versus fats at room temperature. Cheng 2025 did a bang up job characterizing lipids in the atherosclerotic plaques yet did not characterize the cell membrane lipid content that can affect membrane tension/fluidity. This site covers the effect of PEMF on ceramide from acid sphingomyelinase. See the ceramide post.

Wang Y, Li X, Sun L, Feng B, Sun W. Acid sphingomyelinase mediates 50-Hz magnetic field-induced EGF receptor clustering on lipid raft. J Recept Signal Transduct Res. 2016 Dec;36(6):593-600. Pub Med Sci-Hub free

Filamentous actin is a protein polymer that looks and acts like cables maintaining the structure of the cell, often called the cytoskeleton. Ca²⁺regulates many aspects of the cytoskeleton. Cheng 2025 produced some amazing PEMF changes in F-actin in these atherosclerosis models.

Atomic Force Microscopy

The many applications of atomic force microscopy are described in great detail by Wikipedia authors. Nothing in this page suggests that trawling the surface of the cell activates stretch activated ion channels.

Stewart AP, Smith GD, Sandford RN, Edwardson JM. Atomic force microscopy reveals the alternating subunit arrangement of the TRPP2-TRPV4 heterotetramer. Biophys J. 2010 Aug 4;99(3):790-7. PMC free paper

This group was using AFM to map the subunit composition of these heterotetramers. No mention was made if probing these channels resulted in gating. Part of the like of hypertension to atherosclerosis comes from the shear forces on vessel walls. Does raking a atomic force probe across the surface mimic shear forces associated with hypertension?

Electrophysiology

The interesting thing is that the authors are measuring all of the currents in these whole cells in physiological inside and outside solutions. Thing get crazy because intracellular Ca²⁺ can influence the gating of other ion channels including TRP family members via Ca²⁺-calmodulin. A Ca²⁺ binding protein. The types of patch clamping link. Whole cell patch clamping is the easiest even if it gets all ion channels in the cell. Patch clamping during PEMF exposure could probably have been impossible without a Faraday cage shown in this Molecular Devices diagram of a patch clamp rig.

The Mitochondria

The Seahorse is a “respirometer” that measures oxygen consumption at complex IV where O₂ is reduced to H₂0, and “non-mitochondrial” O₂ consumption. Many biohackers are familiar with the “lore” that PEMF boosts the flux of electrons through complex IV and promotes the notion that superoxide generation from active mitochondria is part of normal business and that PEMF might promote ROS generation.

JC-1 is fluorescent compound used to detect losses in the mitochondrial membrane potential that target the mitochondria to be “eaten” by lysosomes. The eating of dysfunctional mitochondria, mitophagy, is considered to be a healthy process as the dysfunctional mitochondria generate reactive oxygen species. Wikipedia’s page on mitophagy is pretty good if you scroll down to the part on Pink1 and Parkin, yes named after Parkinson’s disease. Pink1 accumulates in the mitochondria that have lost their membrane potential, i.e. have become dysfuncional. Parkin comes along and attaches “eat me” ubiquitin tags that target mitocondria to the lysosomes, cellular stomachs. Mitophagy is important in energy demanding tissues like the brain. The Wikipedia have a few things to say about mitophagy and inflammasomes. Mitochondrial DNA can activate inflammasomes.

Inflammasomes and Western Blotting

Wikipedia has a good piece on Western blotting. Basically one homogenizes cells or tissue in a detergent called SDS that you might also be in your shampoo. Proteins in mixtures are separated by size in porous gel slabs by applying a voltage gradient. The SDS gives the proteins a negative, anionic, charge. Anions go to the anode. The smaller the protein, the faster it moves through the gel. The proteins in the gel are transferred to a sicky membrane. Bare spots on the sticky membranes are blocked with a powdered milk colloid. Antibody combinations are used to detect the inflammasome proteins.

Cytokines, components of the inflammasomes, and ion channels were measured based on the binding of antibodies raised in the first animal, like a rabbit, injected with the protein, and then an antibody raised in a second animal, like a goat, against immunoglobins from a rabbit. The anti-rabbit goat antibodies are tagged with a probe for detection. Many times scientists take the easy route and measure the messenger RNA, mRNA, transcripts and assume they get translated into proteins equally. These protein based methods don’t tell us if more protein is the result of more gene transcription and translation, or less degradation. Genomics and proteomics approach in combination are generally required to get the full picture.

Human clinical data

Plasma is blood minus the cells. Cheng 2025 digested the whole proteins with the pancreatic enzyme trypsin. The peptides were separated by a combination of size and charge. The Orbitrap Astral was used to measure the size of the peptides and presumably the size of the fragments when the peptides were collided and allowed to decay into pieces. Proteins can be identified quickly based on the size of the fragments. Databases can be searched to correlate the proteins with a particular function. It almost as if this analysis was part of another study and Cheng 2025 just threw it in to bulk up their study. What really irritates me from a PEMF biohacker perspective is that they could have done the same sort of plasma proteomic analysis to ask if there were any differences in proteins in the ND, HFD, and HFD+PEMF mice. They were exposing the whole mice to PEMF, not just the aorta. Maybe something in the circulation far removed from the aorta was being released by PEMF and then influencing fat deposits in the aorta.

NLRP3 knock down

Short hairpin RNA, shRNA, is a means of silencing target genes, in this case the NLRP3 subunit of inflammasomes. AAV, adeno-associated viruses are used for gene therapy. The mCherry fluorescent probe that goes along with the shRNA and informs the scientists that the transfection was successful. We still have the same problem that streptomycin is a TRP channel inhibitor. I like that knocking down is done in cultured cells. I don’t much like the endothelial cell specific knock down in whole mice.

Experimental cell culture groups

“The human umbilical vein endothelial cell line (CRL-1730) was purchased from the American Type Culture Collection and cultured with Ham’s F12k (Gibco, 21127022, USA) and 10% fetal bovine serum (FBS, Gibco, 26010074, USA) supplemented with 1% penicillin‒streptomycin (HyClone, SV30010, USA). HUVECs were cultured in a cell incubator at 37 °C and 5% CO₂. All experiments were conducted using cells at passages 4–10. The HAEC cell line (IM-H359) was purchased from Xiamen Immocell Biotechnology Co., Ltd. (Fujian, China) and cultured in HAEC-specific medium (Immocell, IM-H359-1, China). HAECs were cultured in a cell incubator at 37 °C and 5% CO₂. All experiments were conducted using cells at passages 4–10.”

Franco-Obregón A, Tai YK. Are Aminoglycoside Antibiotics TRPing Your Metabolic Switches? Cells. 2024 Jul 29;13(15):1273. PMC free paper

“Critically, aminoglycoside antibiotics (AGAs) have been shown to block the capacity of TRP channels to conduct calcium entry into the cell in response to a wide range of developmental stimuli of a biophysical nature, including mechanical, electromagnetic, thermal, and chemical.” And, as near as we care tell, these endothelial cells were not subjected to any kind of mechanical stress.

Experimental mouse groups

The Cherry-NC is a fluorescent protein marker to tell the group that the transfection was in the targeted cells was successful. This seems to be more precise than the double knockout that can affect multiple organ systems. I’m going to assume that the “no drug” control mice got injected with saline such that the stress of handling could be controlled for. Are Nigericin, HC, HY, and GSK safe and non-toxic at these doses? I don’t honestly know. We have to trust the reviewers to know these things. I had to do a key word search to figure out that all the mice were males. It would be interesting to know the lipid composition of this high fat diet as Cheng 2025 performed “lipidomics on the atherosclerotic plaques of these animals. Are the lipids in the plaques what was in the diet in the first place?

Here is an image generated from one of my favorite resources: Proteinatlas.org.

The PEMF device

“The magnetic field device was manufactured by Sichuan University (Fig. S10). The mice in the PEMF group were exposed to PEMFs for 3 weeks after 12 weeks of HFD. PEMFs were generated with the following parameters: adjustable pulse wave, frequency = 15 Hz, intensity = 1.5 mT, pulse duration = 0.3 s, pulse duty ratio: 30%, and exposure time = 60 min/day. The parameters of PEMFs in vitro and in vivo follow the experimental results of previous studies and the parameter gradients used in this study. The parameters of the PEMFs were divided into three groups for the experiments to screen out the optimal parameters

• 1.5 Hz/0.5 mT/60 min/3 weeks (PEMF I group)
• 2.15 Hz/1.5 mT/60 min/3 weeks (PEMF II group),
• 3. 30 Hz/3.0 mT/60 min/3 weeks (PEMF III group).

The in vitro PEMF intervention device was installed in the cell incubator. HUVECs were exposed to PEMFs, and the frequency and intensity (15 Hz, 1.5 mT) were the same as those used for PEMF intervention in vivo.” It would have been nice to have seen a dose that caused toxicity in the cultured cells or insects if a toxic exposure would not be permitted in rodents.

Plasma lipid biochem

1. normal chow diet
2. high fat diet for 15 weeks
3. (PEMF I group) 1.5  Hz/0.5 mT/60 min/3 weeks
4. (PEMF II group) 2.15 Hz/1.5 mT/60 min/3 weeks
5. (PEMF III group) 3.30 Hz/3.0 mT/60 min/3 weeks

Results, by Figure

1. Chicken or the egg PEMFs inhibit the formation of atherosclerotic plaques by attenuating pyroptosis and inflammation. “Which came first: reduction in fat or reduction in inflammation?”
2. monitoring proteins of inflammation in cell culture PEMFs reduce the Ox-LDL-induced inflammatory response and pyroptosis and improve EC survival in an inflammatory environment. 
3. knocking out inflammasomes The bidirectional regulation of NLRP3 downregulation has the same effect on PEMFs, whereas excessive upregulation counteracts the therapeutic effect of PEMFs.
4. Inflammasome subunits Downregulation of NLRP3 can protect cells, and upregulation of NLRP3 can aggravate EC damage.
5. Mitochondrial dysfunction is a bridge between inflammation and atherosclerosis. LDL induced mitochondrial dysfunction in cultured cells. PEMF alleviated some dysfunction. PEMF restored much of the ATP production in HUVEC but had little effect i mouse vascular endothelial cells MVEC.
6. Endothelial cell membrane tension and transient receptor potential vanilloid 4 (TRPV4) mechanosensitive channels participate in Ox-LDL/HFD-induced endothelial damage.
7. TRPV4 hyperactivation exacerbates cellular stiffness and mitochondrial dysfunction, whereas TRPV4 inhibition confers cytoprotection GSK1016790A is a potent TRPV4 agonist whereas HC67047 is an inhibitor.
8. TRPV4 activation exacerbates atherogenesis, whereas TRPV4 inhibition has the same effect on PEMFs. The TRPC4 inhibitor HC067047 was given to mice. For bladder function and blood pressure, a contributing factor for atherosclerosis.

Caveats

The unfortunate twist here is that we cannot trust results obtained in the cultured cells because of gentamycin. The APOE-/- background, the use of whole body exposure of the mice when a human patient may only have the hard to stent affected major vessel exposed?

Causes of aorta stiffening was reviewed in a previous post. Endothelial nitric oxide synthase (eNOS) as an antenna for PEMF was covered in the eNOS & PI3K post. These signaling pathways were not considered in Cheng 2025.