EMF acts on rat bone marrow mesenchymal stem cells to promote differentiation to osteoblasts and to inhibit differentiation to adipocytes

The use of electromagnetic fields (EMFs) to treat nonunion fractures developed from observations in the mid‐1900s. Whether EMF directly regulates the bone marrow mesenchymal stem cells (MSCs), differentiating into osteoblasts or adipocytes, remains unknown. In the present study, we investigated the roles of sinusoidal EMF of 15 Hz, 1 mT in differentiation along these separate lineages using rat bone marrow MSCs. Our results showed that EMF promoted osteogenic differentiation of the stem cells and concurrently inhibited adipocyte formation. EMF increased alkaline phosphatase (ALP) activity and mineralized nodule formation, and stimulated osteoblast‐specific mRNA expression of RUNX2, ALP, BMP2, DLX5, and BSP. In contrast, EMF decreased adipogenesis and inhibited adipocyte‐specific mRNA expression of adipsin, AP‐2, and PPARγ2, and also inhibited protein expression of PPARγ2. These observations suggest that commitment of MSCs into osteogenic or adipogenic lineages is influenced by EMF. Bioelectromagnetics 31:277–285, 2010. © 2009 Wiley‐Liss, Inc.     

Effects of low-intensity AC and/or DC electromagnetic fields on cell attachment and induction of apoptosis

Rat tendon fibroblast (RTF) and rat bone marrow (RBM) osteoprogenitor cells were cultured and exposed to AC and/or DC magnetic fields in a triaxial Helmholtz coil in an incubator for up to 13 days. The AC fields were at 60 and 1000 Hz and up to 0.25 mT peak to peak, and the DC fields were up to 0.25 mT. At various combinations of field strengths and frequencies, AC and/or DC fields resulted in extensive detachment of preattached cells and prevented the normal attachment of cells not previously attached to substrates. In addition, the fields resulted in altered cell morphologies. When RTF and RBM cells were removed from the fields after several days of exposure, they partially reattached and assumed more normal morphologies. An additional set of experiments described in the Appendix corroborates these findings and also shows that low-frequency EMF also initiates apoptosis, i.e., programmed cell death, at the onset of cell detachment. Taken together, these results suggest that the electromagnetic fields result in significant alterations in cell metabolism and cytoskeleton structure. Further work is required to determine the relative effect of the electric and magnetic fields on these phenomena. The research has implications for understanding the role of fields in affecting bone healing in fracture nonunions, in cell detachment in cancer metastasis, and in the effect of EMF on organisms generally. Bioelectromagnetics 18:264–272, 1997. © Wiley-Liss, Inc.     

Effects of low frequency electromagnetic field on proliferation of human epidermal stem cells: An in vitro study

To investigate the effects of low frequency electromagnetic fields (EMF) on the proliferation of epidermal stem cells, human epidermal stem cells (hESC) were isolated, expanded ex vivo, and then exposed to a low frequency EMF. The test and control cells were placed under the same environment. The test cells were exposed for 30 min/day to a 5 mT low frequency EMF at 1, 10, and 50 Hz for 3, 5, or 7 days. The effects of low frequency EMF on cell proliferation, cell cycle, and cell‐surface antigen phenotype were investigated. Low frequency EMF significantly enhanced the proliferation of hESC in the culture medium in a frequency‐dependent manner, with the highest cell proliferation rate at 50 Hz (P < 0.05). Exposure to a low frequency EMF significantly increased the percentage of cells at the S phase of the cell cycle, coupled with a decrease in the percentage of cells in the G1 phase (P < 0.05) but the effect was not frequency dependent. The percentage of CD29⁺/CD71^? cells remained unchanged in the low frequency EMF‐exposed hESC. The results suggested that low frequency EMF influenced hESC proliferation in vitro, and this effect was related to the increased proportion of cells at the S phase. Bioelectromagnetics 34:74–80, 2013. © 2012 Wiley Periodicals, Inc.     

Effects of 50 Hz EMF exposure on micronucleus formation and apoptosis in transformed and nontransformed human cell lines

Effects of applying extremely low-frequency electromagnetic fields (ELF-EMF) for different durations (24, 48, and 72 h) and different field intensities (0.1–1.0 mT) on micronucleus (MN) formation and induction of apoptosis were examined in a human squamous cell carcinoma cell line (SCL II) and in a human amniotic fluid cell line (AFC). A statistically significant increase of MN frequency and of induction of apoptosis in SCL II cells after 48-h and 72-h continuous exposure to 50 Hz magnetic field (MF) (0.8 and 1.0 mT) was found. However, exposure of AFC cells to EMF of different intensities and for different exposure times showed no statistically significant differences when compared with controls. These results demonstrate that different human cell types respond differently to EMF. Dose-dependent induction of apoptosis and genotoxic effects, resulting in increased micronucleus formation, could be demonstrated in the transformed cell line, whereas the nontransformed cell line did not show statistically significant effects. These findings suggest that EMF could be a promotor but not an initiator of carcinogenic effects. Bioelectromagnetics 19:85–91, 1998. © 1998 Wiley-Liss, Inc.     

Effects of extremely low frequency electromagnetic field (EMF) on collagen type I mRNA expression and extracellular matrix synthesis of human osteoblastic cells

Human osteoblastic cells were grown in a three-dimensional (3-D) cell culture model and used to test the effects of a 20 Hz sinusoidal electromagnetic field (EMF; 6 mT and 113 mV/cm max) on collagen type I mRNA expression and extracellular matrix formation in comparison with the effects of growth factors. The cells were isolated from trabecular bone of a healthy individual (HO-197) and from a patient presenting with myositis ossificans (MO-192) and grown in a collagenous sponge-like substrate. Maximal enhancement of collagen type I expression after EMF treatment was 3.7-fold in HO-197 cells and 5.4-fold in MO-192 cells. Similar enhancement was found after transforming growth factor-β (TGF-β) and insulin-like growth factor-I (IGF-I) treatment. Combined treatment of the cells with EMF and the two growth factors TGF-β and IGF-I did not act synergistically. MO-192 cells produced an osteoblast-characteristic extracellular matrix containing collagen type I, alkaline phosphatase, and osteocalcin, together with collagen type III, TP-1, and TP-3, two epitopes of an osteoblastic differentiation marker. The data suggest that the effects of EMFs on osteoblastic differentiation are comparable to those of TGF-β and IGF-I. We conclude that EMF effects in the treatment of skeletal disorders and in orthopedic adjuvant therapy are mediated via enhancement of collagen type I mRNA expression, which may lead to extensive extracellular matrix synthesis. Bioelectromagnetics 19:222–231, 1998. © 1998 Wiley-Liss, Inc.     

Acute exposure to a 60 Hz magnetic field increases DNA strand breaks in rat brain cells

Acute (2 h) exposure of rats to a 60 Hz magnetic field (flux densities 0.1, 0.25, and 0.5 mT) caused a dose-dependent increase in DNA strand breaks in brain cells of the animals (assayed by a microgel electrophoresis method at 4 h postexposure). An increase in single-strand DNA breaks was observed after exposure to magnetic fields of 0.1, 0.25, and 0.5 mT, whereas an increase in double-strand DNA breaks was observed at 0.25 and 0.5 mT. Because DNA strand breaks may affect cellular functions, lead to carcinogenesis and cell death, and be related to onset of neurodegenerative diseases, our data may have important implications for the possible health effects of exposure to 60 Hz magnetic fields. Bioelectromagnetics 18:156–165, 1997. © 1997 Wiley-Liss, Inc.     

Extremely low‐frequency electromagnetic fields affect the immune response of monocyte‐derived macrophages to pathogens

This study aimed to determine the effect of extremely low‐frequency electromagnetic fields (ELF‐EMF) on the physiological response of phagocytes to an infectious agent. THP‐1 cells (human monocytic leukemia cell line) were cultured and 50 Hz, 1 mT EMF was applied for 4–6 h to cells induced with Staphylococcus aureus or interferon gamma/lipopolysaccharide (IFγ/LPS). Alterations in nitric oxide (NO), inducible nitric oxide synthase (iNOS) levels, heat shock protein 70 levels (hsp70), cGMP levels, caspase‐9 activation, and the growth rate of S. aureus were determined. The growth curve of exposed bacteria was lower than the control. Field application increased NO levels. The increase was more prominent for S. aureus‐induced cells and appeared earlier than the increase in cells without field application. However, a slight decrease was observed in iNOS levels. Increased cGMP levels in response to field application were closely correlated with increased NO levels. ELF‐EMF alone caused increased hsp70 levels in a time‐dependent manner. When cells were induced with S. aureus or IFγ/LPS, field application produced higher levels of hsp70. ELF‐EMF suppressed caspase‐9 activation by a small extent. These data confirm that ELF‐EMF affects bacterial growth and the response of the immune system to bacterial challenges, suggesting that ELF‐EMF could be exploited for beneficial uses. Bioelectromagnetics 31:603–612, 2010. © 2010 Wiley‐Liss, Inc.     

Coupling factors and exosomal packaging microRNAs involved in the regulation of bone remodelling

Bone remodelling is a continuous process by which bone resorption by osteoclasts is followed by bone formation by osteoblasts to maintain skeletal homeostasis. These two forces must be tightly coordinated not only quantitatively, but also in time and space, and its malfunction leads to diseases such as osteoporosis. Recent research focusing on the cross‐talk and coupling mechanisms associated with the sequential recruitment of osteoblasts to areas where osteoclasts have removed bone matrix have identified a number of osteogenic factors produced by the osteoclasts themselves. Osteoclast‐derived factors and exosomal‐containing microRNA (miRNA) can either enhance or inhibit osteoblast differentiation through paracrine and juxtacrine mechanisms, and therefore may have a central coupling role in bone formation. Entwined with angiocrine factors released by vessel‐specific endothelial cells and perivascular cells or pericytes, these factors play a critical role in angiogenesis–osteogenesis coupling essential in bone remodelling.     

The regenerative effects of electromagnetic field on spinal cord injury

Traumatic spinal cord injury (SCI) is typically the result of direct mechanical impact to the spine, leading to fracture and/or dislocation of the vertebrae along with damage to the surrounding soft tissues. Injury to the spinal cord results in disruption of axonal transmission of signals. This primary trauma causes secondary injuries that produce immunological responses such as neuroinflammation, which perpetuates neurodegeneration and cytotoxicity within the injured spinal cord. To date there is no FDA-approved pharmacological agent to prevent the development of secondary SCI and induce regenerative processes aimed at healing the spinal cord and restoring neurological function. An alternative method to electrically activate spinal circuits is the application of a noninvasive electromagnetic field (EMF) over intact vertebrae. The EMF method of modulating molecular signaling of inflammatory cells emitted in the extra-low frequency range of <100 Hz, and field strengths of <5 mT, has been reported to decrease inflammatory markers in macrophages, and increase endogenous mesenchymal stem cell (MSC) proliferation and differentiation rates. EMF has been reported to promote osteogenesis by improving the effects of osteogenic media, and increasing the proliferation of osteoblasts, while inhibiting osteoclast formation and increasing bone matrix in vitro. EMF has also been shown to increase chondrogenic markers and collagen and induce neural differentiation, while increasing cell viability by over 50%. As advances are made in stem cell technologies, stabilizing the cell line after differentiation is crucial to SCI repair. Once cell-seeded scaffolds are implanted, EMF may be applied outside the wound for potential continued adjunct treatment during recovery.     

Short‐term exposure to 50 Hz ELF‐EMF alters the cisplatin‐induced oxidative response in AT478 murine squamous cell carcinoma cells

The aim of this study was to assess the influence of cisplatin and an extremely low frequency electromagnetic field (ELF‐EMF) on antioxidant enzyme activity and the lipid peroxidation ratio, as well as the level of DNA damage and reactive oxygen species (ROS) production in AT478 carcinoma cells. Cells were cultured for 24 and 72 h in culture medium with cisplatin. Additionally, the cells were irradiated with 50 Hz/1 mT ELF‐EMF for 16 min using a solenoid as a source of the ELF‐EMF. The amount of ROS, superoxide dismutase (SOD) isoenzyme activity, glutathione peroxidase (GSH‐Px) activity, DNA damage, and malondialdehyde (MDA) levels were assessed. Cells that were exposed to cisplatin exhibited a significant increase in ROS and antioxidant enzyme activity. The addition of ELF‐EMF exposure to cisplatin treatment resulted in decreased ROS levels and antioxidant enzyme activity. A significant reduction in MDA concentrations was observed in all of the study groups, with the greatest decrease associated with treatment by both cisplatin and ELF‐EMF. Cisplatin induced the most severe DNA damage; however, when cells were also irradiated with ELF‐EMF, less DNA damage occurred. Exposure to ELF‐EMF alone resulted in an increase in DNA damage compared to control cells. ELF‐EMF lessened the effects of oxidative stress and DNA damage that were induced by cisplatin; however, ELF‐EMF alone was a mild oxidative stressor and DNA damage inducer. We speculate that ELF‐EMF exerts differential effects depending on the exogenous conditions. This information may be of value for appraising the pathophysiologic consequences of exposure to ELF‐EMF. Bioelectromagnetics 33:641–651, 2012. © 2012 Wiley Periodicals, Inc.     

The Short‐Term Effect of Occupational Levels of 50 Hz Electromagnetic Field on Human Heart Rate Variability

Previous studies have indicated that there is no consensus on the effects of extremely low‐frequency electromagnetic (ELF‐EMF) exposure on the cardiovascular system. This study aimed to explore the short‐term effect of ELF‐EMF exposure on heart rate (HR) and HR variability (HRV). The sample consisted of 34 healthy males aged 18–27 years. The participants were randomly assigned to the EMF (n = 17) or the Sham group (n = 17). We employed a double‐blind repeated‐measures design consisting of three 5 min experimental periods. The chest region of each individual in the EMF group was exposed to 50 Hz, 28 μT, linear polarized, continuous EMF during the EMF exposure period. HR and HRV data were recorded continuously by using a photoplethysmography sensor. Within‐subject statistical analysis indicated a significant HR deceleration in both the EMF and Sham groups. However, the standard deviation of the NN intervals (SDNN), root mean square of successive differences (RMSSD), low‐frequency (LF), and high‐frequency (HF) powers increased only in the EMF group and remained stable in the Sham group. We also compared the same HRV indices measured during the EMF and Sham periods between the two experimental groups. The between‐subject analysis results demonstrated significantly higher SDNN, RMSSD, LF, and HF values in the EMF group than in the Sham group. The LF/HF ratio did not change significantly within and between groups. On the basis of these results, we concluded that short‐term exposure of the chest region to ELF‐EMF could potentially enhance parasympathetic predominance during the resting condition. Bioelectromagnetics. 2021;42:60–75. © 2020 Bioelectromagnetics Society.     

EXPOSURE TO 50 HZ ELECTROMAGNETIC FIELDS INDUCES THE PHOSPHORYLATION AND ACTIVITY OF STRESS-ACTIVATED PROTEIN KINASE IN CULTURED CELLS

Protein phosphorylation is one of the important processes of cell signal transduction pathways. To study the effects of 50 Hz electromagnetic field (EMF) on the cell signal transduction process, the phosphorylation of stress-activated protein kinase (SAPK/JNK) extracted from Chinese hamster lung (CHL) cells exposed to 0.4 and 0.8 mT 50 Hz EMF for various durations was measured. A solid-phase kinase assay was used to measure the enzymatic activity of SAPK extracted from cells exposed to 50 Hz EMF at the same magnetic flux density and for only 15 min. The results showed that both 0.4 and 0.8 mT could induce the phosphorylation of SAPK, the phosphorylation of SAPK presented a time-dependent course, and there was a difference between the two intensities. The phosphorylated SAPK enhanced its enzymatic activity. All the data indicated that 50 Hz EMF could activate SAPK in a time- and intensity-dependent manner. The biological effects caused by 50 Hz EMF maybe related to the SAPK signal transduction pathway.     

Effect of 50 Hz sinusoidal electromagnetic field on the kinetics of 14CO2 exhalation after [14C]‐N‐Nitrosodiethylamine administration in mice

N‐Nitrosodiethylamine (NDEA) has been identified as a typical environmental carcinogen. Its metabolism was studied in mice under the influence of an electromagnetic field (EMF). After intraperitoneal administration of [¹⁴C]‐NDEA, 0.2 μCi/100 g body weight resulted in 22.8% of the total radioactivity exhaled as ¹⁴CO₂ within 1 h. Mice were exposed to a 50 Hz, 2 mT (rms) electromagnetic field, 8 h/day for 8 weeks. There was a significant increase in the metabolic turnover of [¹⁴C]‐NDEA into ¹⁴CO₂ at the end of both 6 and 8 weeks of field exposure, i.e., 26.9% and 37.4% respectively. The enhanced capacity of mice to metabolize NDEA after the exposure to EMF may result in animals with a smaller amount of the bioactive carcinogen burden, thereby indicating a protective role of 2 mT EMF in a whole animal study. Bioelectromagnetics 20:1–4, 1999. © 1999 Wiley‐Liss, Inc.     

Pulsed Electromagnetic Field Versus Whole Body Vibration on Cartilage and Subchondral Trabecular Bone in Mice With Knee Osteoarthritis

Pulsed electromagnetic field (PEMF) and whole body vibration (WBV) interventions are expected to be important strategies for management of osteoarthritis (OA). The aim of the study was to investigate the comparative effectiveness of PEMF versus WBV on cartilage and subchondral trabecular bone in mice with knee OA (KOA) induced by surgical destabilization of the medial meniscus (DMM). Forty 12-week-old male C57/BL mice were randomly divided into four groups (n = 10): Control, OA, PEMF, and WBV. OA was induced (OA, PEMF, and WBV groups) by surgical DMM of right knee joint. Mice in PEMF group received 1 h/day PEMF exposure with 75 Hz, 1.6 mT for 4 weeks, and the WBV group was exposed to WBV for 20 min/day with 5 Hz, 4 mm, 0.3 g peak acceleration for 4 weeks. Micro-computed tomography (micro-CT), histology, and immunohistochemistry analyses were performed to evaluate the changes in cartilage and microstructure of trabecular bone. The bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N) increased, and bone surface/bone volume (BS/BV) decreased by micro-CT analysis in PEMF and WBV groups. The Osteoarthritis Research Society International (OARSI) scores in PEMF and WBV groups were significantly lower than in the OA group. Immunohistochemical results showed that PEMF and WBV promoted expressions of Aggrecan, and inhibited expressions of IL-1β, ADAMTS4, and MMP13. Superior results are seen in PEMF group compared with WBV group. Both PEMF and WBV were effective, could delay cartilage degeneration and preserve subchondral trabecular bone microarchitecture, and PEMF was found to be superior to WBV. Bioelectromagnetics. 2020;41:298–307 © 2020 Bioelectromagnetics Society     

50 Hz 1.8 mT正弦交变电磁场通过调节成骨细胞PGE2的分泌影响Opg/Rankl的基因表达

前列腺素E2(prostaglandin E2, PGE2)作为细胞因子,在骨代谢中扮演重要角色. 它通过刺激成骨细胞核因子κB受体活化因子配基(receptor activator of nuclear factor kappa B ligand, RANKL)表达,促进破骨细胞的分化成熟. 然而,其是否参与了电磁场调节骨代谢仍不清楚.PGE2的生物合成受到环加氧酶(cyclooxygenase, COX)的调节. 在细胞中存在2种不同的环加氧酶,COX-1和COX-2. 其中,COX-2是引起PGE2分泌增加的主要原因. 其活性受到细胞核因子κB(nuclear factor kappa B, NF-κB)的调节.本文通过检测体外培养成骨细胞PGE2分泌,COX-2蛋白表达以及Cox-2、Opg、Rankl和Nf-κb 基因表达发现,经50 Hz 1.8 mT正弦交变电磁场(sinusoidal electromagnetic fields, SEMFs)处理后,由COX-2介导的PGE2分泌以及cox-2、Nf-κb的基因表达皆下调,但Nf-κb的变化先于cox-2的变化,而opg/rankl基因表达则恰恰相反,说明电磁场通过抑制Nf-κb的转录降低由COX-2介导的PGE2的分泌,进而降低对Rankl表达的刺激作用,抑制破骨细胞的分化成熟.     

Membrane trafficking in osteoblasts and osteoclasts: new avenues for understanding and treating skeletal diseases

The endocytic and exocytic/secretory pathways are two major intracellular membrane trafficking routes that regulate numerous cellular functions in a variety of cell types. Osteoblasts and osteoclasts, two major bone cells responsible for bone remodeling and homeostasis, are no exceptions. During the past few years, emerging evidence has pinpointed a critical role for endocytic and secretory pathways in osteoblast and osteoclast differentiation and function. The endosomal membrane provides a platform to integrate bone tropic signals of hormones and growth factors in osteoblasts. In osteoclasts, endocytosis, followed by transcytosis, of degraded bone matrix promotes bone resorption. Secretory pathways, especially lysosome secretion, not only participate in bone matrix deposition by osteoblasts and degradation of mineralized bone matrix by osteoclasts; they may also be involved in the coupling of bone resorption and bone formation during bone remodeling. More importantly, mutations in genes encoding regulatory factors within the endocytic and secretory pathways have been identified as causes for bone diseases. Identification of the molecular mechanisms of these genes in bone cells may provide new therapeutic targets for skeletal disorders.     

Influence of extremely low frequency magnetic fields on chromosomes and the mitotic cycle in Vicia faba L., the broad bean

Vicia faba seedlings were subjected to one of the following magnetic fields continuously for 3 days: 0 Hz (DC) at 5 mT, 50 Hz at 1.5 mT, 60 Hz at 1.5 mT, and 75 Hz at 1.5 mT. The lengths of all the phases of mitosis differed from the controls in all treatments using alternating magnetic fields and for prophase and metaphase in the DC condition. In particular, all treatments increased the length of prophase significantly in meristematic root-tip cells compared with the controls. The implications of these results for chromosome coiling are discussed. The length of prophase, however, did not vary significantly between any of the treatments. Furthermore, none of the exposed seedlings had a greater frequency of chromosome breakages above that of the control plants. Bioelectromagnetics 19:152–161, 1998. © 1998 Wiley-Liss, Inc.     

Interaction between osteoblast and osteoclast: impact in bone disease

The intercellular communication between osteoblasts and osteoclasts is crucial to bone homeostasis. Since Rodan and Martin proposed the control of osteoclasts by osteoblasts in the 1980s, many factors have been isolated from osteoblasts and shown to regulate the differentiation and function of osteoclasts. However, the mechanism by which osteoblasts regulate osteoclasts during bone remodelling is still unclear. On the other hand, it is well accepted that many metabolic bone diseases are associated with the disruption of the communication between osteoblast and osteoclasts. Thus, this review focuses on the cross-talk between osteoblasts and osteoclasts and its impact in bone disease.     

The roles of bone‐derived exosomes and exosomal microRNAs in regulating bone remodelling

Pathological destructive bone diseases are primarily caused by the failure of a lifelong self‐renewal process of the skeletal system called bone remodelling. The mechanisms underlying this process include enhanced osteoclast activity and decreased generation of the osteoblast lineage. Intercellular interaction and crosstalk among these cell types are crucial for the maintenance of bone remodelling, either through the secretion of growth factors or direct cell–cell physical engagement. Recent studies have revealed that exosomes derived from bone cells, including osteoclasts, osteoblasts and their precursors, play pivotal roles on bone remodelling by transferring biologically active molecules to target cells, especially in the processes of osteoclast and osteoblast differentiation. Here, we review the contents of bone‐derived exosomes and their functions in the regulatory processes of differentiation and communication of osteoclasts and osteoblasts. In addition, we highlight the characteristics of microRNAs of bone‐derived exosomes involved in the regulation of bone remodelling, as well as the potential clinical applications of bone‐derived exosomes in bone remodelling disorders.