Electromagnetic Field Stimulation Research Articles

Transcriptomic Analysis of Gene Expression and Effect of Electromagnetic Field in Brain Tissue after Traumatic Brain Injury.

Traumatic brain injury (TBI) due to a direct blow or penetrating injury to the head damages the brain tissue and affects brain function. Primary and secondary damage to the brain tissue increases disability, morbidity, and mortality and costs millions of dollars in treatment. Injury to the brain tissue results in the activation of various inflammatory and repair pathways involving many cellular and molecular factors. Increased infiltration of immune cells to clear the debris and lesion healing, activation of Schwann cells, myelination, oligodendrocyte formation, and axonal regeneration occur after TBI to regenerate the tissue. However, secondary damage to brain tissue results in behavioral symptoms. Repair and regeneration are regulated by a complex cascade involving various cells, hormones, and proteins. A change in the expression of various proteins due to altered gene expression may be the cause of impaired repair and the sequelae in TBI. In this pilot study, we used a Yucatan miniswine model of TBI with and without electromagnetic field (EMF) stimulation and investigated the differential gene expression between injured and non-injured cortex tissues. We found several differentially expressed genes including INSC, TTR, CFAP126, SEMA3F, CALB1, CDH19, and SERPINE1. These genes are associated with immune cell infiltration, myelination, reactive oxygen species regulation, thyroid hormone transportation, cell proliferation, and cell migration. There was a time-dependent effect of EMF stimulation on the gene and protein expression. The findings support the beneficial effect of EMF stimulation in the repair process following TBI.

Transcriptional and Translational Regulation of Differentially Expressed Genes in Yucatan Miniswine Brain Tissues following Traumatic Brain Injury.

Traumatic brain injury (TBI) is a leading cause of morbidity, disability, and mortality worldwide. Motor and cognitive deficits and emotional disturbances are long-term consequences of TBI. A lack of effective treatment for TBI-induced neural damage, functional impairments, and cognitive deficits makes it challenging in the recovery following TBI. One of the reasons may be the lack of knowledge underlying the complex pathophysiology of TBI and the regulatory factors involved in the cellular and molecular mechanisms of inflammation, neural regeneration, and injury repair. These mechanisms involve a change in the expression of various proteins encoded by genes whose expression is regulated by transcription factors (TFs) at the transcriptional level and microRNA (miRs) at the mRNA level. In this pilot study, we performed the RNA sequencing of injured tissues and non-injured tissues from the brain of Yucatan miniswine and analyzed the sequencing data for differentially expressed genes (DEGs) and the TFs and miRs regulating the expression of DEGs using in-silico analysis. We also compared the effect of the electromagnetic field (EMF) applied to the injured miniswine on the expression profile of various DEGs. The results of this pilot study revealed a few DEGs that were significantly upregulated in the injured brain tissue and the EMF stimulation showed effect on their expression profile.

Pulsed electromagnetic field stimulation enhances neurite outgrowth in neural cells and modulates inflammation in macrophages

Nerve regeneration following traumas remains an unmet challenge. The application of pulsed electromagnetic field (PEMF) stimulation has gained traction for a minimally invasive regeneration of nerves. However, a systematic exploration of different PEMF parameters influencing neuron function at a cellular level is not available. In this study, we exposed neuroblastoma F11 cells to PEMF to trigger beneficial effects on neurite outgrowth. Different carrier frequencies, pulse repetition frequencies, and duty cycles were screened with a custom ad hoc setup to find the most influential parameters values. A carrier frequency of 13.5 MHz, a pulse repetition frequency of 20 Hz, and a duty cycle of 10% allowed maximal neurite outgrowth, with unaltered viability with respect to non-stimulated controls. Furthermore, in a longer-term analysis, such optimal conditions were also able to increase the gene expression of neuronal expression markers NeuN and Tuj-1 and transcription factor Ngn1. Finally, the same optimal stimulation conditions were also applied to THP-1 macrophages, and both pro-inflammatory (TNF-α, IL-1β, IL-6, IL-8) and anti-inflammatory cytokines (IL-10, CD206) were analyzed. The optimal PEMF stimulation parameters did not induce differentiation towards an M1 macrophage phenotype, decreased IL-1β and IL-8 gene expression, decreased TNF-α and IL-8 cytokine release in M1-differentiated cells, increased IL-10 and CD206 gene expression, as well as IL-10 cytokine release in M0 cells. The specific PEMF stimulation regime, which is optimal in vitro, might have a high potential for a future in vivo translation targeting neural regeneration and anti-inflammatory action for treating peripheral nerve injuries.

Pulsed Electromagnetic Field Stimulation in Lumbar Spine Fusion for Patients With Risk Factors for Pseudarthrosis.

Lumbar spinal fusion surgeries are increasing steadily due to an aging and ever-growing population. Patients undergoing lumbar spinal fusion surgery may present with risk factors that contribute to complications, pseudarthrosis, prolonged pain, and reduced quality of life. Pulsed electromagnetic field (PEMF) stimulation represents an adjunct noninvasive treatment intervention that has been shown to improve successful fusion and patient outcomes following spinal surgery. A prospective, multicenter study investigated PEMF as an adjunct therapy to lumbar spinal fusion procedures in patients at risk for pseudarthrosis. Patients with at least 1 of the following risk factors were enrolled: prior failed fusion, multilevel fusion, nicotine use, osteoporosis, or diabetes. Fusion status was determined by radiographic imaging, and patient-reported outcomes were also evaluated. A total of 142 patients were included in the analysis. Fusion status was assessed at 12 months follow-up where 88.0% (n = 125/142) of patients demonstrated successful fusion. Fusion success for patients with 1, 2+, or 3+ risk factors was 88.5%, 87.5%, and 82.3%, respectively. Significant improvements in patient-reported outcomes using the Short Form 36, EuroQol 5 Dimension (EQ-5D) survey, Oswestry Disability Index, and visual analog scale for back and leg pain were also observed compared with baseline scores (P < 0.001). A favorable safety profile was observed. PEMF treatment showed a positive benefit-risk profile throughout the 6-month required use period. The addition of PEMF as an adjunct treatment in patients undergoing lumbar spinal surgery provided a high rate of successful fusion with significant improvements in pain, function, and quality of life, despite having risk factors for pseudarthrosis. PEMF represents a useful tool for adjunct treatment in patients who have undergone lumbar spinal surgery. Treatment with PEMF may result in improved fusion and patient-reported outcomes, regardless of risk factors. NCT03176303.

Stimulation of soy seeds using environmentally friendly magnetic and electric fields

The study analyses the impact of alternating (magnetic induction B = 30 mT for t = 60 s) and constant magnetic fields (B = 130 mT for t = 17 h) and alternating electric fields (electric current E = 5 kV/cm for t = 60 s) on various growth parameters of soy plants: the germination energy and capacity, plants emergence, the fresh mass of seedlings, protein content (Kjeldahl’s method), and photosynthetic parameters (with MINI-PAM 2000 WALTZ Photosynthesis Yield Analyser and a SPAD-502 Chlorophyll Meter). Four cultivars were used: MAVKA, MERLIN, VIOLETTA, and ANUSZKA. Moreover, the advanced Machine Learning processing pipeline was proposed to distinguish the impact of physical factors on photosynthetic parameters. The use of electromagnetic fields had a positive impact on the germination rate in MERLIN seeds. The best results in terms of germination improvement were observed for alternating magnetic field stimulation in all cultivars (p > 0.05). For the VIOLETTA cultivar an increase (p > 0.05) in the emergence and overall number of plants as well as fresh mass was observed after electromagnetic field stimulation. For the MAVKA and MERLIN cultivars, the concentration of proteins in the leaves was noticeably higher in plants grown from seeds stimulated using a constant magnetic field.

A Swine Model of Neural Circuit Electromagnetic Fields: Effects of Immediate Electromagnetic Field Stimulation on Cortical Injury.

Background Neurologic diseases have profound disability, mortality, and socioeconomic effects worldwide. Treatment of these disorders varies but is largely limited to unique factors associated with neural physiology. Early studies have evaluated alterations in electromagnetic fields (EMF) due to neural disorders with subsequent modulation of EMF as a potential treatment modality. Swine models have begun to be evaluated as translational models in this effect. Methods EMF measurements of a Yucatan miniswine were recorded using proprietary non-contact, non-invasive induction sensors with a dual layer Mu-metal and interlaced copper mesh helmet. The swine then underwent controlled cortical impact (CCI) to simulate traumatic brain injury (TBI). Twenty minutes post-injuryafter surgical wound closure, the swine underwent targeted EMF signal modulation using a signal generator to stimulate the swine's injured cortical circuit using a sinusoidal wave individualized at 2.5 Hz with a 500mV positive offset at 1V. After 10 days of stimulation, settings were modified to another individualized frequency of 5.5 Hz, 500mV positive offset and 1V for stimulation. Behavioral patterns in swine were evaluated, and EMF measurements were recorded daily prior to, during, and after stimulation. Artificial intelligence (AI) models evaluated patterns in EMF signals.Histology of the stimulated swine cortex was evaluated using hematoxylin and eosin staining and pentachrome staining and compared to a control swine without stimulation and a swine that had received stimulation two days post-injury in a delayed fashion. Serial serum specimens and tissue at the time of euthanasia were obtained for assessment of neuron-specific enolase (NSE) concentration. Results Pre-operative and post-stimulation measurements demonstrated differences in patterns and activity early on. There was an identified peak at 1.6Hz, not frequently seen pre-operatively. There were convergent frequencies in both data setsat 10.5 Hz and 3.9 Hz. Plateaus and decreased variability of changes in slope were identified early in the post-injury phase. AI modeling identified early similarities in pre-operative and post-stimulation measurements through the patterns of peaks with similarities on postoperative day 10 and similarities in the valleys on postoperative day 17.Histologic specimens identified increased degrees of apoptosis and cellular death in the non-stimulated control compared to the stimulated swine. Similarly, the immediately stimulated swine had less apoptosis and increased histologic viability at the site of injury compared to the two-day delayed stimulation swine. There were increased levels of NSE noted in the stimulated swine at the site of injury compared to non-injured sites and the control swine. Conclusions Cortical function was appropriately measured through induction sensors and shielding in the form of a helmet and electromagnetic field channels. Early stimulation resulted in the early and durable recovery of neuronal circuit-driven electromagnetic field patterns. Histology identified increased viability of neurons with fewer apoptotic neurons and glial cells in stimulated swine with early stimulation identifying the best effect compared to a non-stimulated subject. This recovery identifies change and recovery at the circuit, cellular, and subcellular levels that potentiatethe need for further study of EMF modulation as a treatment modality in neurological disorders.

A Swine Model of Traumatic Brain Injury: Effects of Neuronally Generated Electromagnetic Fields and Electromagnetic Field Stimulation on Traumatic Brain Injury-Related Changes.

Background and objective Traumatic brain injury (TBI) has been associated with aberrations in neural circuitry attributable to the pathology resulting in electromagnetic field (EMF) changes. These changes have been evaluated in a variety of settings including through novel induction sensors with an ultra-portable shielded helmet and EMF channels with differences identified by comparing pre-injury and post-injury states. Modulation of the EMF has undergone cursory evaluation in neurologic conditions but has not yet been fully evaluated for clinical effects in treatment. Target EMF stimulation using EMF-related changes preoperatively to postoperatively has not yet been attempted and has not been completed using induction sensor technology. Our objectives in this study were twofold: we wanted to test thehypothesis that targeted stimulation using an EMF signal generator and stimulator to abnormal thresholds identified by real-time measurement of EMFs may enable early resolution of EMF changes and treatment of the TBI as modeled through controlled cortical impact (CCI); we also aimed toassess the feasibility of attemptingthis using real-time measurements with an EMF shielded helmet with EMF channels and non-contact, non-invasive induction sensors with attached EMF transmitters in real-time. Methods A singular Yucatan miniswine was obtained and baseline EMF recordings were obtained. A CCI of TBI and postoperative assessment of cortical EMF in a non-invasive, non-contact fashion were completed. Alterations in EMF were evaluated and EMF stimulation using those abnormal frequencies was completed using multiple treatments involving three minutes of EMF stimulation at abnormal frequencies. Stimulation thresholds of 2.5 Hz, 3.5 Hz, and 5.5 Hz with 1 V signal intensity were evaluated using sinusoidal waves. Additionally, stimulation thresholds using differing offsets to the sine wave at -500 mV, 0 mV, and 500 mv were assessed. Daily EMF and post-stimulation EMF measurements were recorded. EMF patterns were then assessed using an artificial intelligence(AI) model. Results AI modeling appropriately identified differences in EMF signal in pre-injury, post-injury, and post-stimulation states. EMF stimulation using a positive offset of 500 mV appeared to have maximal beneficial effects in return to baseline. Similarly targeted stimulation using thresholds of 2.5 Hz and 5.5 Hz with a positive 500 mV offset at 1 V allowed for recovery of EMF patterns post-injury towards patterns seen in baseline EMF measurements on stimulation day seven (postoperative day 17). Conclusion Stimulation of neural circuits with targeted EMF in a sinusoidal pattern with targeted thresholds after measurement with induction sensors with shielding isolated to a Mu-metal and copper mesh helmet and EMF channels is efficacious in promoting neuronal circuit recovery to preoperative baselines in the TBI miniswine model. Similarly, our findings confirm the appropriateness of this translational model in the evaluation of brain neuronal circuit EMF and that preoperative and post-trauma differences can be appropriately assessed with this technology.

The effect of electromagnetic field stimulation on the electromagnetic spectrum structure of essenstial oil

The effect of electromagnetic field stimulation on the electromagnetic spectrum structure of essenstial oil

Postoperative Rehabilitation to Improve Outcomes After Cervical Spine Fusion for Degenerative Cervical Spondylosis: A Systematic Review.

Postoperative rehabilitation has recently been identified as a high-priority research topic for improving surgical outcomes for degenerative cervical spondylosis (DCS). However, there remains no consensus on specific rehabilitation strategies. Thus, the objective of this study was to evaluate the effectiveness of postoperative rehabilitation strategies for short-term and long-term outcomes after cervical spine fusion forDCS. A systematic review was performed according to Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines using the PubMed, Scopus, and Ovid Medline databases. All level I-IV therapeutic studies in the English language investigating the outcomes of postoperative rehabilitation strategies after cervical spine fusion for DCS were included. Nine studies with 895 patients with DCS (747 anterior-only fusion, 55 patients with posterior-only fusion, 93 patients with physiotherapy alone) were included in this analysis, with 446 (49.8%) patients receiving physiotherapy alone or standard postoperative therapyand 449 (50.2%) patients receiving standard postoperative therapy with additional intervention or augmentation. These interventions included pulsed electromagnetic field (PEMF) stimulation, telephone-supported home exercise program (HEP), early cervical spine stabilizer training, structured postoperative therapy, and a postoperative cervical collar. One level II study demonstrated that PEMF led to increased fusion rates at six months postoperatively compared to standard therapy alone, one level II study demonstrated that postoperative cervical therapy in addition to standard therapy was better than standard therapy alone in the improvement of neck pain intensity, one level IV study demonstrated home exercise therapy led to an improvement in neck pain, arm pain, and disability, and six level II studies reported no difference in clinical outcome measures between augmented or targeted therapy and standard postoperative therapy for DCS. In conclusion, there is moderate evidence to suggest that there is no significant difference in clinical and surgical outcomes between standard postoperative therapy and augmented or targeted postoperative therapy for cervical fusion in the setting of cervical spondylosis. However, there is some evidence to support that certain therapeutic modalities, such as PEMF stimulation, may lead to improved fusion rates, clinical outcomes, and patient satisfaction when compared to standard postoperative therapy protocols. There is no evidence to support a difference in effectiveness with different types of postoperative rehabilitation strategies between anterior and posterior fusions for DCS.

Dynamic orbital hybridization triggered spin-disorder renormalization via super-exchange interaction for oxygen evolution reaction

The oxygen evolution reaction (OER) underpins many aspects of energy storage and conversion in modern industry and technology, but which still be suffering from the dilemma of sluggish reaction kinetics and poor electrochemical performance. Different from the viewpoint of nanostructuring, this work focuses on an intriguing dynamic orbital hybridization approach to renormalize the disordering spin configuration in porous noble-metal-free metal-organic frameworks (MOFs) to accelerate the spin-dependent reaction kinetics in OER. Herein, we propose an extraordinary super-exchange interaction to reconfigure the domain direction of spin nets at porous MOFs through temporarily bonding with dynamic magnetic ions in electrolytes under alternating electromagnetic field stimulation, in which the spin renormalization from disordering low-spin state to high-spin state facilitates rapid water dissociation and optimal carrier migration, leading to a spin-dependent reaction pathway. Therefore, the spin-renormalized MOFs demonstrate a mass activity of 2,095.1 A gmetal-1 at an overpotential of 0.33V, which is about 5.9 time of pristine ones. Our findings provide a insight into reconfiguring spin-related catalysts with ordering domain directions to accelerate the oxygen reaction kinetics.

Stimulasi Pulsed Electromagnetic Field (PEMF) pada Osteogenesis Pergerakan Gigi Ortodonti

The prevalence of malocclusion and the need for orthodontic treatment in Indonesia is relatively very high. Orthodontic treatment that transmits mechanical forces to the periodontal ligament and alveolar bone cells can stimulate the periodontal ligament and alveolar bone remodeling. While this time, orthodontic treatment requires a long time, resulting in several adverse effects and discomfort. For this reason, a method that can accelerate orthodontic treatment is needed which aims to increase stability after active orthodontic treatment, to reduce the potential for relapse, and to accelerate retention time. Pulsed Electromagnetic Field (PEMF) stimulation is one of the methods that have this goal. This aim to study the stimulation mechanism of PEMF on the osteogenesis process of orthodontic tooth movement. Alveolar bone remodeling played an essential role in orthodontic tooth movement, is an active and dynamic process. The dynamic process depends on a balance between resorption and bone formation (osteogenesis). PEMF stimulation is a nonsurgical method that can promote osteogenesis through activation of Wnt signaling. Several studies showed that this method could accelerate retention time and orthodontic treatment. PEMF stimulated osteogenesis through activation of Wnt signaling in orthodontic tooth movement.

Electromagnetic investigation of neuron growth by using pulsed electromagnetic field stimulation

Electromagnetic field (EMF) applications for humans have been widely used for medical therapy for a long time. Various nerve diseases such as complex regional pain syndrome, cerebral palsy and spinal cord injuries may occur as a result of damage on the nerve cells. In addition, EMF applications are taken into consideration in terms of tissue regeneration. The use of electricity for nerve regeneration is applied internally or externally. The use of direct current (DC) electric fields to stimulate axon elongation and nerve regeneration in nerve cells has been applied in various studies. In our study, electromagnetic field was used for the controlled and directional growth of Dorsal Root Ganglia (DRG) nerve cells. An amplifier circuit, which can produce amplified signals to generate proposed pulsed electromagnetic fields (PEMF) is designed. The electromagnetic field is produced on FR-4 copper plate, which highlights electrical field unlike previous studies, since there is no circular coil structure preferred. The elongation of nerve cells is planned to be in one direction, hence the signal to be applied is produced as a novel asymmetrical waveform (sawtooth) in addition to triangular and square waveforms which were previously preferred in recent studies. By the aid of proposed novel construction and produced signal waveform, PEMF system can be applied for nerve diseases effectively. The morphological nerve cell elongation results and PEMF results have been investigated and the effectiveness of the proposed system is verified.

Applications of Pulsed Electromagnetic Field Therapy in Skeletal-Muscle System: An Integrative Review

Background: The Pulsed Electromagnetic Field (PEMF) technology has attracted increasing interest since consistent evidence of its therapeutic properties has been demonstrated to treat musculoskeletal conditions. However, this technology is not new, and has already been used in different experimental models and clinical studies for the treatment of tendinopathies, osteoarthritis, increased cell proliferation, bone consolidation, among others. Methods: In this work, we carried out an integrative review of clinical and experimental studies published in the last twenty years, on the available scientific evidence that demonstrate the effects of PEMF in different applications for health treatments. Five databases including Medline, Pubmed Central, Scopus, Lilacs and PEDro were searched for studies from 2001 to September 2022. The results were analyzed by the team of researchers and clinical professionals to assure methodological quality of the studies for the elaboration of the theoretical review on the effects of electromagnetic field stimulation on skeletal muscles, tendon or bones. Results: Sixty-two studies were included in this review, presenting evidence of the biological effects of PEMF that can suggest it´s possible use to treat different disorders. Conclusions: Pulsed electromagnetic fields (PEMF) present relevant clinical and experimental evidence of beneficial effects in the treatment of several musculoskeletal inflammatory disorders, such as tendinopathies and osteoarthritis, in addition to the treatment of urinary incontinence and abdominal diastasis. PEMF can be considered as the evolution of electrical currents for the treatment of musculoskeletal disorders, mainly due to its better tolerance by patients.

Numerical Modeling and Computer Simulation of a Meander Line Antenna for Alzheimer's Disease Treatment, a Feasibility Study.

Alzheimer's disease (AD) is a brain disorder that eventually causes memory loss and the ability to perform simple cognitive functions; research efforts within pharmaceuticals and other medical treatments have minimal impact on the disease. Our preliminary biological studies showed that Repeated Electromagnetic Field Stimulation (REFMS) applying an EM frequency of 64 MHz and a specific absorption rate (SAR) of 0.4 - 0.9 W/kg decrease the level of amyloid-β peptides (Aβ), which is the most likely etiology of AD. This study emphasizes uniform E/H field and SAR distribution with adequate penetration depth penetration through multiple human head layers driven with low input power for safety treatments. In this work, we performed numerical modeling and computer simulations of a portable Meander Line antenna (MLA) to achieve the required EMF parameters to treat AD. The MLA device features a low cost, small size, wide bandwidth, and the ability to integrate into a portable system. This study utilized a High-Frequency Simulation System (HFSS) in the design of the MLA with the desired characteristics suited for AD treatment in humans. The team designed a 24-turn antenna with a 60 cm length and 25 cm width and achieved the required resonant frequency of 64 MHz. Here we used two numerical human head phantoms to test the antenna, the MIDA and spherical head phantom with six and seven tissue layers, respectively. The antenna was fed from a 50-Watt input source to obtain the SAR of 0.6 W/kg requirement in the center of the simulated brain tissue layer. We found that the E/H field and SAR distribution produced was not homogeneous; there were areas of high SAR values close to the antenna transmitter, also areas of low SAR value far away from the antenna. This paper details the antenna parameters, the scattering parameters response, the efficiency response, and the E and H field distribution; we presented the computer simulation results and discussed future work for a practical model.

Disease-Modifying Effects of Non-Invasive Electroceuticals on β-Amyloid Plaques and Tau Tangles for Alzheimer’s Disease

Electroceuticals refer to various forms of electronic neurostimulators used for therapy. Interdisciplinary advances in medical engineering and science have led to the development of the electroceutical approach, which involves therapeutic agents that specifically target neural circuits, to realize precision therapy for Alzheimer's disease (AD). To date, extensive studies have attempted to elucidate the disease-modifying effects of electroceuticals on areas in the brain of a patient with AD by the use of various physical stimuli, including electric, magnetic, and electromagnetic waves as well as ultrasound. Herein, we review non-invasive stimulatory systems and their effects on β-amyloid plaques and tau tangles, which are pathological molecular markers of AD. Therefore, this review will aid in better understanding the recent technological developments, applicable methods, and therapeutic effects of electronic stimulatory systems, including transcranial direct current stimulation, 40-Hz gamma oscillations, transcranial magnetic stimulation, electromagnetic field stimulation, infrared light stimulation and ionizing radiation therapy, and focused ultrasound for AD.

Effects of the Pulsed Electromagnetic Fields on Experimental Periodontitis and Estrogen Deficiency.

Periodontitis is an inflammatory disease resulting from a complex polymicrobial infection that causes tissue destruction in susceptible individuals. Osteoporosis has been associated with greater clinical attachment loss in patients with periodontitis. Experimental studies have shown positive results in the treatment of osteoporosis through pulsed electromagnetic field (PEMF) stimulation. The aim of this study was to evaluate the effects of PEMF in the presence of estrogen deficiency associated with periodontitis, verifying its role in bone metabolism and in the inflammatory response. Sixty rats were divided into four groups: Sham surgery + ligature-induced periodontitis (P); Sham surgery + ligature-induced periodontitis + PEMF therapy (P + PEMF); Ovariectomy surgery + ligature-induced periodontitis (P + OVX); Ovariectomy surgery + ligature-induced periodontitis + PEMF therapy (P + OVX + PEMF). The area of bone loss in the furcation region (BL), connective tissue attachment loss (CTAL) and alveolar bone loss (ABL), BV/TV and BMD were evaluated. In addition to immunohistochemical labelling of RANKL, OPG, and TRAP and the inflammatory response of interleukin (IL)-1b, IL-6, TNF-α, IL-10, and vascular endothelial growth factor. P + OVX showed significant BL in relation to P + PEMF and the greatest CTAL and ABL. P + OVX and P + OVX + PEMF showed a significant reduction in BV/TV (%). P and P + PEMF showed a significantly lesser amount of Tb.Sp (mm) while P + OVX and P + OVX + PEMF showed a lesser of Tb.N. P + PEMF had the greatest BMD. P + OVX presented higher RANKL and lower OPG immunolabeling than other groups. P + PEMF and P + OVX + PEMF showed a reduction on all biomarkers evaluated. The application of PEMF seems to attenuate the effects of bone loss in the presence of periodontitis and ovariectomy. © 2022 Bioelectromagnetics Society.

P17.17.A Metformin, tmCLIC1 and transcranial stimulation: the basis of a new targeted therapy against glioblastoma relapse

Abstract Background Glioblastoma (GBM) is the most common brain tumor with an extremely poor prognosis. Standard therapy relies on surgery, chemo- and radiotherapy but tumor relapse occurs inevitably with fatal outcome. Hence, new approaches should be designed to increase patients’ life expectancy. The high grade of aggressiveness and recurrence is attributed to GBM stem-like cells (GSCs). Recent publications propose the transmembrane Chloride Intracellular Channel 1 protein (tmCLIC1) as a potential pharmacological target since it is crucial for GSCs proliferation both in vitro and in vivo. Recently, we proposed tmCLIC1 to be one of metformin’s targets. Here, we show molecular evidence of direct metformin-tmCLIC1 interaction. However, the high concentration of metformin needed to impair tumor progression cannot be reached in the brain due to the blood brain barrier. Hence, the main aim of this work is to decrease metformin’s operative concentration to the level that can access the brain. Our strategy is to induce repetitive membrane depolarizations to the tumoral area to promote metformin-tmCLIC1 interaction, that only occurs when tmCLIC1 is in the open state. Pulsed depolarizing electromagnetic field (EMF) stimulation should increase tmCLIC1 close-to-open transitions and, consequently, the availability of metformin binding sites. Material and Methods We generated CLIC1-knockout of several patient-derived GSCs using CRISPR-Cas9 technology to assess the role of CLIC1 in metformin’s antiproliferative effect. In the first set of experiments, we used 10mM metformin. When applying concomitant EMF stimulation, we dropped to 1mM metformin. NMR experiments and single-channel electrophysiological recordings were provided to study metformin-tmCLIC1 interaction. Metformin-EMF system was applied in vitro on GSCs cultures and spheroids, and in vivo on zebrafish embryos and mice. Results NMR and electrophysiology demonstrate the direct binding between metformin and tmCLIC1, that figures as the main metformin receptor in GBM. The application of EMF stimulation resulted in a 10-fold decrease of metformin concentration to get the same antiproliferative effect in vitro. To assess whether the effect is maintained in vivo, zebrafish embryos were orthotopically injected with GSCs and the tumor mass was measured after 72 hours in absence or presence of metformin in embryos’ water and of EMF stimulation. Consistently, metformin coupled to EMF reduces tumor progression in zebrafish embryos to the same extent of metformin 10mM alone. Experiments on mice are currently on-going. Conclusion Our research has revealed a new interactor of metformin. The combination of metformin and EMF results in an effective strategy to impair GBM progression both in vitro and in vivo. The long-term goal is to combine transcranial stimulation and metformin administration to patients as an adjuvant therapy. AIRC#24758

An automated 3D-printed perfusion bioreactor combinable with pulsed electromagnetic field stimulators for bone tissue investigations

In bone tissue engineering research, bioreactors designed for replicating the main features of the complex native environment represent powerful investigation tools. Moreover, when equipped with automation, their use allows reducing user intervention and dependence, increasing reproducibility and the overall quality of the culture process. In this study, an automated uni-/bi-directional perfusion bioreactor combinable with pulsed electromagnetic field (PEMF) stimulation for culturing 3D bone tissue models is proposed. A user-friendly control unit automates the perfusion, minimizing the user dependency. Computational fluid dynamics simulations supported the culture chamber design and allowed the estimation of the shear stress values within the construct. Electromagnetic field simulations demonstrated that, in case of combination with a PEMF stimulator, the construct can be exposed to uniform magnetic fields. Preliminary biological tests on 3D bone tissue models showed that perfusion promotes the release of the early differentiation marker alkaline phosphatase. The histological analysis confirmed that perfusion favors cells to deposit more extracellular matrix (ECM) with respect to the static culture and revealed that bi-directional perfusion better promotes ECM deposition across the construct with respect to uni-directional perfusion. Lastly, the Real-time PCR results of 3D bone tissue models cultured under bi-directional perfusion without and with PEMF stimulation revealed that the only perfusion induced a ~ 40-fold up-regulation of the expression of the osteogenic gene collagen type I with respect to the static control, while a ~ 80-fold up-regulation was measured when perfusion was combined with PEMF stimulation, indicating a positive synergic pro-osteogenic effect of combined physical stimulations.

Enhancing Myoblast Fusion and Myotube Diameter in Human 3D Skeletal Muscle Constructs by Electromagnetic Stimulation.

Skeletal muscle tissue engineering (SMTE) aims at the in vitro generation of 3D skeletal muscle engineered constructs which mimic the native muscle structure and function. Although native skeletal muscle is a highly dynamic tissue, most research approaches still focus on static cell culture methods, while research on stimulation protocols indicates a positive effect, especially on myogenesis. A more mature muscle construct may be needed especially for the potential applications for regenerative medicine purposes, disease or drug disposition models. Most efforts towards dynamic cell or tissue culture methods have been geared towards mechanical or electrical stimulation or a combination of those. In the context of dynamic methods, pulsed electromagnetic field (PEMF) stimulation has been extensively used in bone tissue engineering, but the impact of PEMF on skeletal muscle development is poorly explored. Here, we evaluated the effects of PEMF stimulation on human skeletal muscle cells both in 2D and 3D experiments. First, PEMF was applied on 2D cultures of human myoblasts during differentiation. In 2D, enhanced myogenesis was observed, as evidenced by an increased myotube diameter and fusion index. Second, 2D results were translated towards 3D bioartificial muscles (BAMs). BAMs were subjected to PEMF for varying exposure times, where a 2-h daily stimulation was found to be effective in enhancing 3D myotube formation. Third, applying this protocol for the entire 16-days culture period was compared to a stimulation starting at day 8, once the myotubes were formed. The latter was found to result in significantly higher myotube diameter, fusion index, and increased myosin heavy chain 1 expression. This work shows the potential of electromagnetic stimulation for enhancing myotube formation both in 2D and 3D, warranting its further consideration in dynamic culturing techniques.

Effect of Pulsed Electro Magnetic Field versus Transcutaneous Electrical Nerve Stimulation on Menstrual Distress of Primary Dysmenorrhea

Effect of Pulsed Electro Magnetic Field versus Transcutaneous Electrical Nerve Stimulation on Menstrual Distress of Primary Dysmenorrhea