Sinusoidal Electromagnetic Fields Research Articles

EMF treatment delays mesenchymal stem cells senescence during long-term in vitro expansion by modulating autophagy.

Bone marrow mesenchymal stem cells (BMSCs) are widely used in tissue engineering and regenerative medicine as seed cells. Due to low amount in bone marrow, BMSCs must be expanded and cultured in vitro before application. However, the senescence of stem cell caused by long-term in vitro culture greatly limits its efficacy of transplantation. In this study, we propose an approach based on electromagnetic fields (EMF) treatment to rejuvenate aged BMSCs due to long-term in vitro culture. Aged BMSCs were treated with sinusoidal EMF (50Hz, 0.4mT), and stem cell senescence, cell proliferation, cell differentiation, cell stemness and autophagy level were detected. Additionally, aged BMSCs-laden hydrogels were transplanted into the rat critical-sized calvarial defect with or without EMF treatment. The bone formation was evaluated 8weeks after surgery. Our results indicated that the BMSCs age significantly after long-term in vitro passaging. The self-renew, multiple differentiation capacity, senescence phenotypes and stemness of aged BMSCs are partly reversed by EMF treatment with a frequency of 50Hz and strength of 0.4mT. Moreover, declined autophagy level is observed in BMSCs during long-term in vitro passaging and BMSCs senescence is closely associated with autophagy regulation. Additionally, the mechanistic investigation reveals that EMF treatment rejuvenate senescent BMSCs by enhancing autophagy. Furthermore, EMF treatment significantly promote the therapeutic effect of long-term passaged BMSCs on bone formation in vivo. Overall, our study identifies a practical approach for the rejuvenation of old BMSCs and may provide a promising candidate in tissue engineering and stem cell therapy.

A novel implant surface modification mode of Fe3O4-containing TiO2 nanorods with sinusoidal electromagnetic field for osteoblastogenesis and angiogenesis.

Implants made of Ti and its alloys are widely utilized in orthopaedic surgeries. However, insufficient osseointegration of the implants often causes complications such as aseptic loosening. Our previous research discovered that disordered titanium dioxide nanorods (TNrs) had satisfactory antibacterial properties and biocompatibility, but TNrs harmed angiogenic differentiation, which might retarded the osseointegration process of the implants. Magnetic nanomaterials have a certain potential in promoting osseointegration, electromagnetic fields within a specific frequency and intensity range can facilitate angiogenic and osteogenic differentiation. Therefore, this study used Fe3O4 to endow magnetism to TNrs and explored the regulation effects of Ti, TNrs, and Fe3O4-TNrs under 1​mT 15​Hz sinusoidal electromagnetic field (SEMF) on osteoblastogenesis, osseointegration, angiogenesis, and its mechanism. We discovered that after the addition of SEMF treatment to VR-EPCs cultured on Fe3O4-TNrs, the calcineurin/NFAT signaling pathway was activated, which then reversed the inhibitory effect of Fe3O4-TNrs on angiogenesis. Besides, Fe3O4-TNrs with SEMF enhanced osteogenic differentiation and osseointegration. Therefore, the implant modification mode of Fe3O4-TNrs with the addition of SEMF could more comprehensively promote osseointegration and provided a new idea for the modification of implants.

EMF promote BMSCs differentiation and functional recovery in hemiparkinsonian rats

Bone marrow mesenchymal stem cells (BMSCs) have self-renewal ability while maintaining the proliferation facility. The BMSCs reproducing ability could affect by electromagnetic fields (EMFs) as a physical inducing factor. We focused on the EMF (400 µT, 75 Hz) exposed multi-potential BMSCs which differentiated and successfully implanted in the substantia nigra pars compacta (SNpc) of Parkinson's disease rat model. The purified BMSCs are exposed to sinusoidal and square waveform EMF (1 h/1 week or 7 h/1 day) then injected into the left SNpc of Parkinson’s rats. To evaluate the morphology of EMF exposed BMSCs, the cresyl violet staining labeled the Nissl bodies. After evaluation of the rat's activity by behavioral tests (open-field and rotarod tests), the brains were obtained for the preparation of SNpc blocks and carry out the cresyl violet staining. Cell morphology proved most cell differentiation to neurons in the sinusoidal EMF groups. In the sinusoidal EMF exposure groups, large and small neurons were seen with apparent synapses. Although in the square EMF exposed groups some neurons were seen, most of the differentiated cells were astrocytes, microglia, and oligodendrocyte. The results confirmed an improvement in locomotors' activity of BMSC alone and sinusoidal EMF exposed groups. We presented a low-frequency EMF (75 Hz) to promote the capability of BMSC proliferation, differentiation to neurons and glial cells, and motor coordination activity in the treatment of hemiparkinsonian rats.

Integrated Micro-Devices for a Lab-in-Organoid Technology Platform: Current Status and Future Perspectives.

Advancements in stem cell technology together with an improved understanding of in vitro organogenesis have enabled new routes that exploit cell-autonomous self-organization responses of adult stem cells (ASCs) and homogenous pluripotent stem cells (PSCs) to grow complex, three-dimensional (3D), mini-organ like structures on demand, the so-called organoids. Conventional optical and electrical neurophysiological techniques to acquire functional data from brain organoids, however, are not adequate for chronic recordings of neural activity from these model systems, and are not ideal approaches for throughput screenings applied to drug discovery. To overcome these issues, new emerging approaches aim at fusing sensing mechanisms and/or actuating artificial devices within organoids. Here we introduce and develop the concept of the Lab-in-Organoid (LIO) technology for in-tissue sensing and actuation within 3D cell aggregates. This challenging technology grounds on the self-aggregation of brain cells and on integrated bioelectronic micro-scale devices to provide an advanced tool for generating 3D biological brain models with in-tissue artificial functionalities adapted for routine, label-free functional measurements and for assay’s development. We complete previously reported results on the implementation of the integrated self-standing wireless silicon micro-devices with experiments aiming at investigating the impact on neuronal spheroids of sinusoidal electro-magnetic fields as those required for wireless power and data transmission. Finally, we discuss the technology headway and future perspectives.

Electromagnetic Field as a Treatment for Cerebral Ischemic Stroke.

Cerebral stroke is a leading cause of death and adult-acquired disability worldwide. To this date, treatment options are limited; hence, the search for new therapeutic approaches continues. Electromagnetic fields (EMFs) affect a wide variety of biological processes and accumulating evidence shows their potential as a treatment for ischemic stroke. Based on their characteristics, they can be divided into stationary, pulsed, and sinusoidal EMF. The aim of this review is to provide an extensive literature overview ranging from in vitro to even clinical studies within the field of ischemic stroke of all EMF types. A thorough comparison between EMF types and their effects is provided, as well as an overview of the signal pathways activated in cell types relevant for ischemic stroke such as neurons, microglia, astrocytes, and endothelial cells. We also discuss which steps have to be taken to improve their therapeutic efficacy in the frame of the clinical translation of this promising therapy.

Evaluation of the Sensing Potential of Stem Cell-Secreted Proteins via a Microchip Device under Electromagnetic Field Stimulation.

Most bone tissue engineering models fail to demonstrate the complex cellular functions of living bone; therefore, most translational studies on bone tissue are performed in live models. To reduce the need for live models, we developed a stimulated microchip model for monitoring protein secretion during osteogenesis using human mesenchymal stem cells (hMSCs). We established a bone microchip system for monitoring the in vitro differentiation and sensing the secreted proteins of hMSCs under a sinusoidal electromagnetic field (SEMF), which ameliorates bone healing in a biomimetic natural bone matrix. A 3 V-1 Hz SEMF biophysically stimulated osteogenesis by activating ERK-1/2 and promoting phosphorylation of p38 MAPK kinases. Exposure to a 3 V-1 Hz SEMF upregulated the expression of osteogenesis-related genes and enhanced the expression of key osteoregulatory proteins. We identified 23 proteins that were differentially expressed in stimulated human bone marrow mesenchymal stem cell secretomes or were absent in the control groups. Our on-chip stimulation technology is easy to use, versatile, and nondisruptive and should have diverse applications in regenerative medicine and cell-based therapies.

The frequency window effect of sinusoidal electromagnetic fields in promoting osteogenic differentiation and bone formation involves extension of osteoblastic primary cilia and activation of protein kinase A.

Electromagnetic fields (EMFs) have emerged as a versatile means for osteoporosis treatment and prevention. However, its optimal application parameters are still elusive. Here, we optimized the frequency parameter first by cell culture screening and then by animal experiment validation. Osteoblasts isolated from newborn rats (ROBs) were exposed 90 min/day to 1.8 mT SEMFs at different frequencies (ranging from 10 to 100 Hz, interval of 10 Hz). SEMFs of 1.8 mT inhibited ROB proliferation at 30, 40, 50, 60 Hz, but increased proliferation at 10, 70, 80 Hz. SEMFs of 10, 50, and 70 Hz promoted ROB osteogenic differentiation and mineralization as shown by alkaline phosphatase (ALP) activity, calcium content, and osteogenesis-related molecule expression analyses, with 50 Hz showing greater effects than 10 and 70 Hz. Treatment of young rats with 1.8 mT SEMFs at 10, 50, or 100 Hz for 2 months significantly increased whole-body bone mineral density (BMD) and femur microarchitecture, with the 50 Hz group showing the greatest effect. Furthermore, 1.8 mT SEMFs extended primary cilia lengths of ROBs and increased protein kinase A (PKA) activation also in a frequency-dependent manner, again with 50 Hz SEMFs showing the greatest effect. Pretreatment of ROBs with the PKA inhibitor KT5720 abolished the effects of SEMFs to increase primary cilia length and promote osteogenic differentiation/mineralization. These results indicate that 1.8 mT SEMFs have a frequency window effect in promoting osteogenic differentiation/mineralization in ROBs and bone formation in growing rats, which involve osteoblast primary cilia length extension and PKA activation.

Sinusoidal electromagnetic fields accelerate bone regeneration by boosting the multifunctionality of bone marrow mesenchymal stem cells

BackgroundThe repair of critical-sized bone defects is always a challenging problem. Electromagnetic fields (EMFs), used as a physiotherapy for bone defects, have been suspected to cause potential hazards to human health due to the long-term exposure. To optimize the application of EMF while avoiding its adverse effects, a combination of EMF and tissue engineering techniques is critical. Furthermore, a deeper understanding of the mechanism of action of EMF will lead to better applications in the future.MethodsIn this research, bone marrow mesenchymal stem cells (BMSCs) seeded on 3D-printed scaffolds were treated with sinusoidal EMFs in vitro. Then, 5.5 mm critical-sized calvarial defects were created in rats, and the cell scaffolds were implanted into the defects. In addition, the molecular and cellular mechanisms by which EMFs regulate BMSCs were explored with various approaches to gain deeper insight into the effects of EMFs.ResultsThe cell scaffolds treated with EMF successfully accelerated the repair of critical-sized calvarial defects. Further studies revealed that EMF could not directly induce the differentiation of BMSCs but improved the sensitivity of BMSCs to BMP signals by upregulating the quantity of specific BMP (bone morphogenetic protein) receptors. Once these receptors receive BMP signals from the surrounding milieu, a cascade of reactions is initiated to promote osteogenic differentiation via the BMP/Smad signalling pathway. Moreover, the cytokines secreted by BMSCs treated with EMF can better facilitate angiogenesis and osteoimmunomodulation which play fundamental roles in bone regeneration.ConclusionIn summary, EMF can promote the osteogenic potential of BMSCs and enhance the paracrine function of BMSCs to facilitate bone regeneration. These findings highlight the profound impact of EMF on tissue engineering and provide a new strategy for the clinical treatment of bone defects.

Low-frequency electromagnetic fields combined with tissue engineering techniques accelerate intervertebral fusion

BackgroundIntervertebral fusion is the most common surgery to treat lumbar degenerative disease (LDD). And the graft material used in the operation is derived from the iliac crest to promote fusion. However, autografts possess the fatal disadvantage of lack of source. Therefore, economical and practical bone substitutes are urgently needed to be developed. Sinusoidal electromagnetic fields (EMF) combined with tissue engineering techniques may be an appropriate way to promote intervertebral fusion.MethodsIn this research, porous scaffolds made of polycaprolactone (PCL) and nano-hydroxyapatite (nHA) were used as cell carriers. Then, the scaffolds loaded with bone marrow mesenchymal stem cells (BMSCs) were treated with sinusoidal electromagnetic field and the osteogenic capability of BMSCs was tested later. In addition, an intervertebral disc of the tail vertebra of the rat was removed to construct a spinal intervertebral fusion model with a cell-scaffold implanted. The intervertebral fusion was observed and analyzed by X-ray, micro-CT, and histological methods.ResultsBMSCs stimulated by EMF possess splendid osteogenic capability under an osteogenic medium (OM) in vitro. And the conditioned medium of BMSCs treated with EMF can further promote osteogenic differentiation of the primitive BMSCs. Mechanistically, EMF regulates BMSCs via BMP/Smad and mitogen-activated protein kinase (MAPK)-associated p38 signaling pathways. In vivo experiments revealed that the scaffold loaded with BMSCs stimulated by EMF accelerated intervertebral fusion successfully.ConclusionIn summary, EMF accelerated intervertebral fusion by improving the osteogenic capacity of BMSCs seeded on scaffolds and might boost the paracrine function of BMSCs to promote osteogenic differentiation of the homing BMSCs at the injured site. EMF combined with tissue engineering techniques may become a new clinical treatment for LDD.

First Identification of the Effects of Low Frequency Electromagnetic Field on the Micromolecular Changes in Adipose Tissue-Derived Mesenchymal Stem Cells by Fourier Transform Infrared Spectroscopy.

Purpose:In this study, we hypothesize that exposure of adipose tissue-mesenchymal stem cells (AT-MSCs) to electromagnetic field (EMF) may impact adipose stem cells' micromolecular structure (analyzed using Fourier transform infrared spectroscopy [FTIR]).Materials and Methods:The AT-MSCs were exposed to continuous vertically applied sinusoidal EMF with a frequency of 50 Hz and a flux density of 1.5 mT for 24, 48, and 72 h. After an appropriate time (24, 48, 72 h) cells were washed with PBS, scrubbed, and immediately taken into FTIR analyses.Results:EMFs affect AT-MSCs. The greatest differences were in the range of nucleic acids and proteins in the fingerprint region which occurred after 24 and 48 h of EMF exposure. However, in the case of 72 h of EMF exposure, no significant differences were noticed in the FTIR spectra towards the control.Conclusions:FTIR spectra show differences between samples under the influence of EMF before they will be manifested at the morphological level. The largest differences in the range of nucleic acids and proteins in the fingerprint region occurred at 24 and 48 h of EMF exposure. That means it was during the first 48 h after EMF exposure a great number of dynamic changes occurred. However, in the case of AT-MSCs in 72 h EMF and 72 h control, no significant differences were noted in the FTIR spectra, which means that the chemical composition in these two cases is similar. EMF is not neutral for stem cells, especially in the in the first hours of interaction (24 h, 48 h).

EMF frequency dependent differentiation of rat bone marrow mesenchymal stem cells to astrocyte cells

The numerous factors regulate the bone marrow mesenchymal stem cell (BMMSC) self-renewal and differentiation response. We aimed to analyze the influence of electromagnetic field (EMF) as an external inducing factor on rat BMMSC differentiation and proliferation to neuron and astrocyte cells. BMMSCs extracted from the rats femurs and tibias and incubated in a cell-cultured CO2 incubator. After the third passages, the plates selected randomly and then divided into seven groups (Sham exposed, three groups of square, and three groups of sinusoidal waveform EMF (25, 50, and 75 Hz, 400 μT, 1 h/day). The BMMSCs exposed to EMF at the middle of a Helmholtz coil for 7 days. The viable cell counting and proliferation performed by the MTT test and BMMSC differentiation into the neuron and the astrocyte cell was studied by immunocytochemistry staining.The results confirmed BMMSC viability and proliferation rate reduction in sinusoidal 25 Hz, square 50 Hz and sinusoidal 75 Hz EMF groups compare to sham. The maximum BMMSC differentiation to neuron was considered in sinusoidal 50 Hz and 75 Hz EMF groups. The increase of BMMSC differentiation to astrocyte cell was frequency dependent and the most differentiation was shown in square 75 Hz, and sinusoidal 75 Hz EMF groups. In conclusion, the results suggest that both square and sinusoidal EMF could affect BMMSC development and differentiation to neuron and astrocyte cells. Further studies for the consequence of EMF with wider flux density and frequency on BMMSC are recommended.

Effects and Mechanisms of Exogenous Electromagnetic Field on Bone Cells: A Review.

Osteoporosis, fractures, and other bone diseases or injuries represent serious health problems in modern society. A variety of treatments including drugs, surgeries, physical therapies, etc. have been used to prevent or delay the progression of these diseases/injuries with limited effects. Electromagnetic field (EMF) has been used to non-invasively treat bone diseases, such as fracture and osteoporosis, for many years. However, because a variety of cellular and molecular events can be affected by EMF with various parameters, the precise bioeffects and underlying mechanisms of specific EMF on bone cells are still obscure. Here, we summarize the common therapeutic parameters (frequency and intensity) of major types of EMF used to treat bone cells taken from 32 papers we selected from the PubMed database published in English from 1991 to 2018. Briefly, pulse EMF promotes the proliferation of osteoblasts when its frequency is 7.5-15 Hz or 50-75 Hz and the intensity is 0.40-1.55 mT or 3.8-4 mT. Sinusoidal EMF, with 0.9-4.8 mT and 45-60 Hz, and static magnetic field with 0.1-0.4 mT or 400 mT, can promote osteoblast differentiation and maturation. Finally, we summarize the latest advances on the molecular signaling pathways influenced by EMF in osteoblasts and osteoclasts. A variety of molecules such as adenosine receptors, calcium channels, BMP2, Notch, Wnt1, etc., can be influenced by EMF in osteoblasts. For osteoclasts, EMF affects RANK, NF-κB, MAPK, etc. We speculate that EMF with different frequencies and intensities exert distinct bioeffects on specific bone cells. More high-quality work is required to explore the detailed effects and underlying mechanisms of EMF on bone cells/skeleton to optimize the application of EMF on bone diseases/injuries. Bioelectromagnetics. 2020;41:263-278 © 2020 Bioelectromagnetics Society.

The combinatory effect of sinusoidal electromagnetic field and VEGF promotes osteogenesis and angiogenesis of mesenchymal stem cell-laden PCL/HA implants in a rat subcritical cranial defect

BackgroundRestoration of massive bone defects remains a huge challenge for orthopedic surgeons. Insufficient vascularization and slow bone regeneration limited the application of tissue engineering in bone defect. The effect of electromagnetic field (EMF) on bone defect has been reported for many years. However, sinusoidal EMF (SEMF) combined with tissue engineering in bone regeneration remains poorly investigated.MethodsIn the present study, we investigated the effect of SEMF and vascular endothelial growth factor (VEGF) on osteogenic and vasculogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs). Furthermore, pretreated rBMSC- laden polycaprolactone-hydroxyapatite (PCL/HA) scaffold was constructed and implanted into the subcritical cranial defect of rats. The bone formation and vascularization were evaluated 4 and 12 weeks after implantation.ResultsIt was shown that SEMF and VEGF could enhance the protein and mRNA expression levels of osteoblast- and endothelial cell-related markers, respectively. The combinatory effect of SEMF and VEGF slightly promoted the angiogenic differentiation of rBMSCs. The proteins of Wnt1, low-density lipoprotein receptor-related protein 6 (LRP-6), and β-catenin increased in all inducted groups, especially in SEMF + VEGF group. The results indicated that Wnt/β-catenin pathway might participate in the osteogenic and angiogenic differentiation of rBMSCs. Histological evaluation and reconstructed 3D graphs revealed that tissue-engineered constructs significantly promoted the new bone formation and angiogenesis compared to other groups.ConclusionThe combinatory effect of SEMF and VEGF raised an efficient approach to enhance the osteogenesis and vascularization of tissue-engineered constructs, which provided a useful guide for regeneration of bone defects.

Effects of sinusoidal electromagnetic fields at different exposure times on osteoblastic activity and the underlying signal transduction mechanisms

Objective To investigate the signal transduction mechanisms of time-dependent effects of 50 Hz 1.8 mT sinusoidal electromagnetic fields(SEMFs)on osteoblastic activity. Methods Newborn rat calvarial osteoblasts(ROBs)were treated with 50 Hz 1.8mT SEMFs for 30, 60, 90, 120, and 150 min, respectively.Intracellular alkaline phosphatase(ALP)activity was assayed, and protein expression of Runt-related transcription factor 2(Runx2), Smad1/5/8 and mitogen-activated protein kinases(MAPKs)were examined by Western blot.The Dual-Luciferase Reporter Assay System was used to measure the activity of the Wnt pathway.Immunofluorescence staining and laser confocal microscopy were applied to examine the nuclear translocation of Smad1/5/8 and β-catenin.Changes in ALP activity were determined after inhibiting p38 MAPK using a specific inhibitor. Results ALP activity of ROBs increased after 30, 60, 90, 120 and 150 min of SEMFs treatment, compared with the control group(51.41±5.21, 59.47±4.02, 67.56±4.68, 63.69±3.92, and 58.16±3.61 vs.45.53±3.24), and reached the highest value at 90 min and then started to decline.Protein expression of Runx-2, p-Smad1/5/8, p-p38 and β-catenin increased after SEMFs treatment, and reached the highest value at 90 min and then gradually returned to baseline levels.The values for Wnt pathway activity for the control group and with SEMFs treatment at 30, 60, 90, 120 and 150 min were 0.49±0.06, 0.52±0.09, 0.75±0.05, 0.77±0.42, 0.58±0.08 and 0.42±0.09, respectively.Wnt pathway activity, the nuclear translocation of β-catenin and Smad1/5/8 reached the highest level at 90 min.Values of ALP activity in the control group and the SEMFs group were 44.60±3.84 and 71.54±7.34, respectively, before specifically inhibiting p38 MAPK, and were 52.08±0.83 and 52.15±10.77, respectively, after p38 MAPK inhibition, indicating that ALP activity could not be increased with inhibition. Conclusions 50 Hz 1.8 mT SEMFs increase osteoblastic activity by activating the BMP-2/Smad1/5/8, Wnt/β-catenin and p38 MAPK signal pathways.The optimal duration of treatment is 90 min per day. Key words: Electromagnetic fields; Osteoblasts; Alkaline phospohatase; Signal transduction

Non-pulsed Sinusoidal Electromagnetic Field Rescues Animals From Severe Ischemic Stroke via NO Activation.

Despite the high prevalence and devastating outcome, only a few treatment options for cerebral ischemic stroke exist. Based on the nitric oxide (NO)-stimulating capacity of Non-pulsed Sinusoidal Electromagnetic Field (NP-SEMF) and the possible neuroprotective role of NO in ischemic stroke, we hypothesized that NP-SEMF is able to enhance survival and neurological outcome in a rat model of cerebral ischemia. The animals, in which ischemic injury was induced by occlusion of both common carotid arteries, received 20 min of NP-SEMF of either 10 or 60 Hz daily for 4 days. NP-SEMF dramatically increased survival, reduced the size of the infarcted brain area and significantly improved the neurological score of the surviving rats. Corresponding to previous reports, NP-SEMF was able to induce NO production in vitro. The importance of NO as a key signaling molecule was highlighted by inhibition of the NP-SEMF beneficial effects in the rat stroke model after blocking NO synthase (NOS). Our results indicate for the first time that NP-SEMF exposure (13.5 mT at 60 and 10 Hz) improves the survival and neurological outcome of rats subjected to cerebral ischemia and that this effect is mediated by NO, underlining the great therapeutic potential of NP-SEMF as a therapy for ischemic stroke.

Human Gingival Fibroblasts Exposed to Extremely Low-Frequency Electromagnetic Fields: In Vitro Model of Wound-Healing Improvement.

Several clinical studies have suggested the impact of sinusoidal and pulsed electromagnetic fields in quickening wound repair processes and tissue regeneration. The clinical use of extremely low-frequency electromagnetic fields could represent a novel frontier in tissue repair and oral health, with an interesting clinical perspective. The present study aimed to evaluate the effect of an extremely low-frequency sinusoidal electromagnetic field (SEMF) and an extremely low-frequency pulsed electromagnetic field (PEMF) with flux densities of 1 mT on a model of oral healing process using gingival fibroblasts. An in vitro mechanical injury was produced to evaluate wound healing, migration, viability, metabolism, and the expression of selected cytokines and protease genes in fibroblasts exposed to or not exposed to the SEMF and the PEMF. Interleukin 6 (IL-6), transforming growth factor beta 1 (TGF-β), metalloproteinase 2 (MMP-2), monocyte chemoattractant protein 1 (MCP-1), inducible nitric oxide synthase (iNOS), and heme oxygenase 1 (HO-1) are involved in wound healing and tissue regeneration, favoring fibroblast proliferation, chemotaxis, and activation. Our results show that the exposure to each type of electromagnetic field increases the early expression of IL-6, TGF-β, and iNOS, driving a shift from an inflammatory to a proliferative phase of wound repair. Additionally, a later induction of MMP-2, MCP-1, and HO-1 was observed after electromagnetic field exposure, which quickened the wound-healing process. Moreover, electromagnetic field exposure influenced the proliferation, migration, and metabolism of human gingival fibroblasts compared to sham-exposed cells. This study suggests that exposure to SEMF and PEMF could be an interesting new non-invasive treatment option for wound healing. However, additional studies are needed to elucidate the best exposure conditions to provide the desired in vivo treatment efficacy.

The Response of Osteoblasts and Bone to Sinusoidal Electromagnetic Fields: Insights from the Literature.

Electromagnetic fields (EMFs) have been proposed as a tool to ameliorate bone formation and healing. Despite their promising results, however, they have failed to enter routine clinical protocols to treat bone conditions where higher bone mass has to be achieved. This is no doubt also due to a fundamental lack of knowledge and understanding on their effects and the optimal settings for attaining the desired therapeutic effects. This review analysed the available in vitro and in vivo studies that assessed the effects of sinusoidal EMFs (SEMFs) on bone and bone cells, comparing the results and investigating possible mechanisms of action by which SEMFs interact with tissues and cells. The effects of SEMFs on bone have not been as thoroughly investigated as pulsed EMFs; however, abundant evidence shows that SEMFs affect the proliferation and differentiation of osteoblastic cells, acting on multiple cellular mechanisms. SEMFs have also proven to increase bone mass in rodents under normal conditions and in osteoporotic animals.

Sinusoidal Electromagnetic Fields Increase Peak Bone Mass in Rats by Activating Wnt10b/β-Catenin in Primary Cilia of Osteoblasts

Extremely low-frequency electromagnetic fields have been considered a potential candidate for the prevention and treatment of osteoporosis; however, their action mechanism and optimal magnetic flux density (intensity) parameter are still elusive. The present study found that 50-Hz sinusoidal electromagnetic fields (SEMFs) at 1.8 mT increased the peak bone mass of young rats by increasing bone formation. Gene array expression studies with femoral bone samples showed that SEMFs increased the expression levels of collagen-1α1 and Wnt10b, a critical ligand of the osteogenic Wnt/β-catenin pathway. Consistently, SEMFs promoted osteogenic differentiation and maturation of rat calvarial osteoblasts (ROBs) in vitro through activating the Wnt10b/β-catenin pathway. This osteogenesis-promoting effect of SEMFs via Wnt10b/β-catenin signaling was found to depend on the functional integrity of primary cilia in osteoblasts. When the primary cilia were abrogated by small interfering RNA (siRNA) targeting IFT88, the ability of SEMFs to promote the osteogenic differentiation of ROBs through activating Wnt10b/β-catenin signaling was blocked. Although the knockdown of Wnt10b expression with RNA interference had no effect on primary cilia, it significantly suppressed the promoting effect of SEMFs on osteoblastic differentiation/maturation. Wnt10b was normally localized at the bases of primary cilia, but it disappeared (or was released) from the cilia upon SEMF treatment. Interestingly, primary cilia were elongated to different degrees by different intensities of 50-Hz SEMFs, with the window effect observed at 1.8 mT, and the expression level of Wnt10b increased in accord with the lengths of primary cilia. These results indicate that 50-Hz 1.8-mT SEMFs increase the peak bone mass of growing rats by promoting osteogenic differentiation/maturation of osteoblasts, which is mediated, at least in part, by Wnt10b at the primary cilia and the subsequent activation of Wnt/β-catenin signaling. © 2019 American Society for Bone and Mineral Research.

Manipulating Mesenchymal Stem Cells Differentiation Under Sinusoidal Electromagnetic Fields Using Intracellular Superparamagnetic Nanoparticles.

In this study, hollow mesoporous ferrite nanoparticles (HMFNs) were prepared. It showed a spherical morphology with a diameter about 320 nm, with a negatively charged surface, and with a great superparamagnetic property. Negative charge attribute to the free -OH group of the HMFNs shell, which improved nanoparticles hydrophilic and biocompatibility. Superparamagnetic property could avoids particle agglomeration. The particles were shown to be internalized into the bone marrow mesenchymal stem cells (BMSCs) in vitro. We found that the intracellular HMFNs can improve the osteogenic differentiation of BMSCs in the presence of an electromagnetic fields. To determine the optimal intensity of the sinusoidal electromagnetic field (SEMF), the exposure levels of 50 Hz SEMF in the range of 0∼4 mTs (60 min/day) were utilized to investigate its effects on the proliferation and osteogenic differentiation of rat BMSCs. The result showed that the 1 mT and 2 mT SEMF stimulated the BMSCs proliferation significantly. The internalized HMFNs in conjunction with SEMF exposure enhanced the osteogenic differentiation, as evidenced by elevated alkaline phosphatase activity, calcium deposition, and the expression protein levels of the expression profile of osteopontin, osteocalcin and runt-related transcription factor 2. We believe that the electromagnetic fields can manipulate osteogenic differentiation of BMSCs using intracellular superparamagnetic nanoparticles.

High inductive magnetic stimuli and their effects on mesenchymal stromal cells, dendritic cells, and fibroblasts.

Effects of low-frequency electromagnetic fields (LF EMF) on the activation of different tissue recovery processes have already been fully understood. Preliminary recent data demonstrated that a special case of sinusoidal electromagnetic fields, known as amplitude-modulated currents (AMC) could have a potential to accelerate the cell metabolism or cell migration. An AMC generator was designed to generate sinusoidal induced electric currents with the amplitude modulation and the harmonic carrier frequency of 5,000 Hz was modulated by frequencies of 1 to 100 Hz. The magnetic field peak was 6 mT, electric field intensity 2 V/m and the current density of induced electrical currents was approximately 1 A/m(2). The coil of the generator was adapted to easy handling and safe integration into the shelf of the CO(2) incubator. The shelf with the coil was prepared for the introduction of cells in standard plastic in vitro chambers. The tests focused on cells with migratory capacity after injury or during immunological processes and thus, mesenchymal stromal cells (MSC), dendritic cells (DC), and fibroblasts were chosen. The tests involved exposures of the cells to LF EMF (180 min/day) every day, for a period of three days, before examining them for cell death, morphology changes, and CD markers. The samples were tested by using MTT assay and the effects on the intracellular concentration of reactive oxygen species were quantified. The cell migration was finally measured with the help of the transwell migration assay. None of the cell types showed any decrease in the cell viability after the LF EMF application and the cells displayed minimum changes in reactive oxygen species. Functional changes (acceleration of cell migration) after AMC exposure were statistically significant for the MSC samples only. The acceleration of MSCs is associated with the production of MMP by these cells. The EMF has a potential to be a safe, clinically applicable selective activator of MSC homing, MSC paracrine production, and subsequent regeneration processes.