Electroporation is a promising technique for enhancing the delivery of therapeutic agents by transiently increasing cell membrane permeability through high-amplitude (from hundreds of V/cm to tens of kV/cm), ultra-short pulsed electric fields (from ms to ns). This technique has shown potential not only for nucleic acid and small molecule delivery but also for the efficient modulation of tumor targets. In this study, we explore, for the first time, the combination of electroporation with an engineered monoclonal antibody targeting tenascin-C, a molecular hallmark of glioblastoma. Using a three-dimensional spheroid model of glioblastoma, we demonstrate that electroporation can enhance antibody binding to tenascin-C in the extracellular matrix, overcoming the structural barriers of the tumor microenvironment. Our results show that electroporation increases antibody access to tumor cells without significant cytotoxicity, highlighting its role as a potentiating tool for antibody-based therapies. Additionally, we utilize plant molecular farming to produce biologically active antibodies, offering a scalable and sustainable platform for large-scale production. This study provides a novel strategy for therapeutic delivery in solid tumors, such as glioblastoma, and offers novel possibilities for other types of cancer.

The global rollout of 5G networks has raised questions regarding the potential biological effects of millimeter-wave exposure, particularly in the skin due to its limited penetration depth. This study examined the effects of whole-body exposure to 27.5 GHz millimeter waves, a frequency within the 5G FR2 bands, on skin-related biological responses in early life mice. Patched1- heterozygous knockout and wild-type CD1 mice were exposed from birth to weaning (P21), 23 h per day, in 10-minute ON/5-minute OFF cycles, at two power densities (6.67 and 20 W/m²). SHAM-exposed animals served as a control. No overt histological abnormalities were observed in exposed skin. However, molecular analyses revealed significant modulation of inflammation-related gene expression. Notably, Ccl4, Csf2, and Tnfsf11 emerged as central regulatory nodes, displaying high degree and betweenness centrality across all groups, irrespective of genotype and sex. Crucially, exposure significantly stimulated mast cell degranulation and, in wild-type mice, led to a reduction in cutaneous glutamate levels. Concurrently, a down-regulation of transcripts associated with cutaneous sensory components (Calca, Mrgprd) was observed within the skin microenvironment. These findings show that 27.5 GHz exposure induces coordinated changes in cutaneous inflammatory pathways and mast cell-mediated homeostasis without detectable structural damage. Overall, these results demonstrate a localized molecular and cellular response within the cutaneous microenvironment, reflecting a subtle homeostatic shift, and suggest that genetic background may contribute to variability in the biological response to millimeter-wave exposure.

Repetitive transcranial magnetic stimulation (rTMS) is an emerging non-invasive therapeutic approach to slow down cognitive and functional decline in Alzheimer’s disease (AD), potentially through plasticity-related mechanisms. MicroRNAs (miRNAs) play a crucial role in synaptic plasticity, and their deregulation contributes to AD-related cognitive impairment. In the present study, we first used a dosimetric model to translate rTMS field applied in AD patients to an in vitro system, identifying miRNAs as potential biomarkers responsive to rTMS. We found that rTMS induced in vitro deregulation of miR-26b, miR-125b, miR-181c, and miR-146a. Then, we investigated the effects of rTMS over precuneus during a 3-week, randomized, sham-controlled trial in AD patients. In patient serum, miR-26b, miR-30b, and miR-125b were significantly modulated in AD patients compared to healthy controls, though no significant modulation emerged between sham and rTMS groups before or after stimulation. Subsequently, the correlation analyses, which incorporated patients’ cognitive scores, revealed that reduced miR-25 levels were significantly associated with cognitive improvement. However, no significant differences emerged between Real- and sham-rTMS correlation coefficients, likely due to the limited sample size, indicating that miR-25 may represent a general prognostic marker rather than a treatment-specific indicator. Furthermore, the ability of this miRNA to discriminate responders from non-responders, shown by ROC analysis, highlights its potential as a promising predictor of rTMS treatment efficacy to be validated in a larger patient cohort. Altogether, our findings suggest, for the first time, that rTMS may modulate specific miRNAs in AD patients, with miR-25 representing a pivotal key target for future validation studies.

Pulsed Electric Fields (PEFs) have emerged as a minimally invasive and highly controllable therapeutic modality for cancer treatment, capable of inducing reversible or irreversible electroporation depending on pulse parameters. Recent advances in Artificial Intelligence (AI) offer new opportunities to optimize PEF protocols by exploiting complex, nonlinear relationships between electrical parameters, cellular properties, and biological outcomes. In this work, we present a novel AI-based framework for the optimization of standalone PEF therapeutic protocols. Unlike standard approaches relying on literature-derived parameter selection, the proposed methodology is grounded on experimental in vitro data obtained from six human tumor cell lines with distinct molecular profiles exposed to various PEF conditions. Multiple Artificial Neural Network (ANN) models were specifically developed to predict cell viability and proliferation and optimize the electrical signal characteristics. The results demonstrate the ability of AI-driven models to accurately capture PEF-induced cellular responses and to guide the selection of effective, tumor-specific therapeutic protocols, paving the way toward data-driven and personalized oncological therapies.

In this work, we used deep learning solutions to predict optimized parameters for pulsed field ablation treatments. This procedure is a new electromagnetic technique used to non-thermally ablate arrhythmogenic cardiac tissue, reducing the side effects associated with microwave thermal ablation. The precision of the lesion, induced by irreversible electroporation of the cardiac tissue, is determined by a large number of parameters (both electromagnetic and biological) that define the pulsed field ablation protocol. The purpose of our work is to predict the various parameters that define pulsed field ablation protocols in an optimized manner using artificial intelligence models. We demonstrated that deep learning methods can optimize and predict these parameters, reducing the need for computationally expensive simulations or trial-anderror approaches, which are both time-consuming and costly. The predictions were based on current literature, and the final estimated accuracy is sufficient to suggest that this approach is promising for the development of future patient-specific protocols.

In the tissue regeneration field, stem cell transplantation represents a promising therapeutic strategy. To favor their implantation, proliferation and differentiation need to be controlled. Several studies have demonstrated that stem cell fate can be controlled by applying continuous electric field stimulation. This study aims to characterize the effect of a specific microsecond electric pulse stimulation (bipolar pulses of 100 µs + 100 µs, delivered for 30 min at an intensity of 250 V/cm) to induce an increase in cell proliferation on mesenchymal stem cells (MSCs) and induced neural stem cells (iNSCs). The effect was evaluated in terms of (i) cell counting, (ii) cell cycle, (iii) gene expression, and (iv) apoptosis. The results show that 24 h after the stimulation, cell proliferation, cell cycle, and apoptosis are not affected, but variation in the expression of specific genes involved in these processes is observed. These results led us to investigate cell proliferation until 72 h from the stimulation, observing an increase in the iNSCs number at this time point. The main outcome of this study is that the microsecond electric pulses can modulate stem cell proliferation.

Objective.Recent studies have indicated that repetitive transcranial magnetic stimulation (rTMS) could enhance cognition in Alzheimer's Disease (AD) patients, but to now the molecular-level interaction mechanisms driving this effect remain poorly understood. While cognitive scores have been the primary measure of rTMS effectiveness, employing molecular-based approaches could offer more precise treatment predictions and prognoses. To reach this goal, it is fundamental to assess the electric field (E-field) and the induced current densities (J) within the stimulated brain areas and to translate these values toin vitrosystems specifically devoted in investigating molecular-based interactions of this stimulation.Approach.This paper offers a methodological procedure to guide dosimetric assessment to translate the E-field induced in humans (in a specific pilot study) intoin vitrosettings. Electromagnetic simulations on patients' head models and cellular holders were conducted to characterize exposure conditions and determine necessary adjustments forin vitroreplication of the same dose delivered in humans using the same stimulating coil.Main results.Our study highlighted the levels of E-field andJinduced in the target brain region and showed that the computed E-field andJwere different among patients that underwent the treatment, so to replicate the exposure to thein vitrosystem, we have to consider a range of electric quantities as reference. To match the E-field to the levels calculated in patients' brains, an increase of at least the 25% in the coil feeding current is necessary whenin vitrostimulations are performed. Conversely, to equalize current densities, modifications in the cells culture medium conductivity have to be implemented reducing it to one fifth of its value.Significance.This dosimetric assessment and subsequent experimental adjustments are essential to achieve controlledin vitroexperiments to better understand rTMS effects on AD cognition. Dosimetry is a fundamental step for comparing the cognitive effects with those obtained by stimulating a cellular model at an equal dose rigorously evaluated.

Electroporation has been shown to have considerable potential in biological research and clinical applications. This technique permits the opening of reversible or irreversible nanometric pores in cellular membranes by the application of a high-intensity electric field characterized by extremely short duration (i.e., microseconds, nanoseconds down to tens of picoseconds). Recently, also thanks to the spreading use of lab-on-chip, it was discovered that applying a high electric field to single cells can cause them to contract and generate blebs. Even though these effects are known in the literature, their correlation with the electroporation phenomenon is still unclear. For this reason, in this work, we present a reliable, low-cost sensing system based on a lab-on-chip platform with transferred laser-induced graphene (LIG) electrodes to study electroporation by bright-field and fluorescence time-lapse microscopy. The acquired video was then used to extrapolate the calcium intake due to electroporation, cell contractility, and morphology over time. The two abovementioned side effects were finally correlated with electroporation and used to estimate its efficacy on single cells, exploiting calcium intake information as the ground truth. The proposed sensing system could be used to predict the electroporation phenomenon easily and even in a label-free modality, using exclusively the morphological variations induced in the single cells.

Objective: Recent studies have indicated that repetitive Transcranial Magnetic Stimulation (rTMS) could enhance cognition in Alzheimer's Disease (AD) patients, yet the molecular-level interaction mechanisms driving this effect remain poorly understood. While cognitive scores have been the primary measure of rTMS effectiveness, employing molecular-based approaches could offer more precise treatment predictions and prognoses. To reach this goal, it is fundamental to assess the electric field (E-field) and the induced current densities (J) within the stimulated brain areas and to translate these values to in vitro systems specifically devoted in investigating molecular-based interactions of this stimulation. Approach: This paper offers a methodological procedure to guide dosimetric assessment to translate the E-field induced in humans (in a specific pilot study) into in vitro settings. Electromagnetic (EM) simulations on patients’ head models and cellular holders were conducted to characterize exposure conditions and determine necessary adjustments for in vitro replication of the same dose delivered in humans using the same stimulating coil. Main results: Our study highlighted the levels of E-field and J induced in the target brain region and showed that the computed E-field and J were different among patients that underwent the treatment, so to replicate the exposure to the in vitro system, we have to consider a range of electric quantities as reference. To match the E-field to the levels calculated in patients’ brains, an increase of at least the 25% in the coil feeding current is necessary when in vitro stimulations are performed. Conversely, to equalize current densities, modifications in the cells culture medium conductivity have to be implemented reducing it to one fifth of its value. Significance: This dosimetric assessment and subsequent experimental adjustments are essential to achieve controlled in vitro experiments to better understand rTMS effects on AD cognition. Dosimetry is a fundamental step for comparing the cognitive effects with those obtained by stimulating a cellular model at an equal dose rigorously evaluated.

In recent years, the interest in transcranial magnetic stimulation (TMS) has surged, necessitating deeper understanding, development, and use of low-frequency (LF) numerical dosimetry for TMS studies. While various ad hoc dosimetric models exist, commercial software tools like SimNIBS v4.0 and Sim4Life v7.2.4 are preferred for their user-friendliness and versatility. SimNIBS utilizes unstructured tetrahedral mesh models, while Sim4Life employs voxel-based models on a structured grid, both evaluating induced electric fields using the finite element method (FEM) with different numerical solvers. Past studies primarily focused on uniform exposures and voxelized models, lacking realism. Our study compares these LF solvers across simplified and realistic anatomical models to assess their accuracy in evaluating induced electric fields. We examined three scenarios: a single-shell sphere, a sphere with an orthogonal slab, and a MRI-derived head model. The comparison revealed small discrepancies in induced electric fields, mainly in regions of low field intensity. Overall, the differences were contained (below 2% for spherical models and below 12% for the head model), showcasing the potential of computational tools in advancing exposure assessment required for TMS protocols in different bio-medical applications.

Spinal Cord Injury (SCI), a major cause of paralysis, currently has no effective therapies. Every year almost 500.000 people are diagnosed with SCI worldwide. he difficulty on the neuronal restoration after SCI is based on the complex cascade of events that inexorably cause a degenerative chronic stage mainly favored by the non-permissive environment and limited capacity for axonal regrowth. RISEUP, an Europen project, proposes to attain neuronal functional regeneration after SCI by an unprecedented and unique bio-hybrid-compatible electro-activated and wireless rechargeable implantable technology. RISEUP introduces high voltage microsecond electric pulses stimulations and low amplitude direct currents on a combination of stem cells, induced neural stem cells (iNSCs) and multipotent stromal cells (MSCs), whose transplantation is facilitated by an innovative scaffold biomaterial (Fig.1).

Spinal cord injury (SCI) is a disabling, devasting and irreversible condition that strongly impacts on patients' life and their families. Every year among 250.000 and 500.000 new SCI cases are diagnosed. Most of them are traumatic and the average of age at diagnosis is 20–29 years for males, while females are most at risk in adolescence (15–19), which means a tremendous socioeconomic impact on affected individuals and the health care system. Recent advances in medical management of SCI have significantly improved diagnosis, stabilization, survival rate and well-being of SCI patients. However, there has been small progress on treatment options for improving the neurological outcomes of SCI patients. This is principally due to the high complexity of spinal cord tissue and to all events that occur after its lesion. RISEUP is a H2020 FET-Open project (nr. 964562) aimed at introducing stem cells microsecond (µs) electric pulses stimulation as a strategy for guiding their differentiation towards a neuronal phenotype for SCI regeneration. Here the Authors report the first results on molecular response induced by µs pulses on mesenchymal stem cells (MSCs) and induced neuronal stem cells (iNSCs).

In recent years, the application of pulsed electric fields with very short durations (nanoseconds) and extremely high amplitudes (MV/m) has been investigated for novel medical purposes. Various electric protocols have been explored for different objectives, including the utilization of fractionated pulse doses to enhance cell electrosensitization to the uptake of different markers or an increase in apoptosis. This study focused on the use of fluorescence imaging to examine molecular calcium fluxes induced by different fractionated protocols of short electric pulses in neuroblastoma (SH-SY5Y) and mesenchymal stem cells (HaMSCs) that were electroporated using nanosecond pulsed electric fields. In our experimental setup, we did not observe cell electrosensitization in terms of an increase in calcium flux following the administration of fractionated doses of nanosecond pulsed electric fields with respect to the non-fractionated dose. However, we observed the targeted activation of calcium-dependent genes (c-FOS, c-JUN, EGR1, NURR-1, β3-TUBULIN) based on the duration of calcium flux, independent of the instantaneous levels achieved but solely dependent on the final plateau reached. This level of control may have potential applications in various medical and biological treatments that rely on calcium and the delivery of nanosecond pulsed electric fields.

Accelerated ripening through the exposure of fruits to controlled environmental conditions and gases is nowadays one of the most assessed food technologies, especially for climacteric and exotic products. However, a fine granularity control of the process and consequently of the quality of the goods is still missing, so the management of the ripening rooms is mainly based on qualitative estimations only. Following the modern paradigms of Industry 4.0, this contribution proposes a non-destructive RFID-based system for the automatic evaluation of the live ripening of avocados. The system, coupled with a properly trained automatic classification algorithm based on Support Vector Machines (SVMs), can discriminate the stage of ripening with an accuracy greater than 85%.

Bio-integrated wireless sensors in the form of conformable plaster, based on the Radiofrequency Identification (RFID) communication, have been recently proposed for the battery-less measurement of the human skin temperature. However, the response of the Integrated Circuit (IC) transponder is sensitive to the strength of the interrogating power. Indeed, high power produces artifacts on the sampled temperature up to 2 °C when the mutual position between reader and sensors, as well as the emitted power, can not be carefully controlled. Hence, a reliable adoption of this technology in real cases is challenging and still in question. A combined macro-scale electromagnetic-thermal model is here introduced to predict and correct the above artifact so that the temperature measurement becomes insensitive to the RF power collected by the IC. The method is based on the new generation RFID ICs with on-chip temperature sensor that are also capable to give back the strength of the collected RF power. The model is validated in controlled conditions and then applied for different skin temperature measurements on human body. An average accuracy of ±0.25 °C, compared with a reference calibrated thermocouple, was demonstrated in the considered tests.

Neuroprotective effects of pulsed electromagnetic fields (PEMFs) have been demonstrated both in vivo and in vitro. Moreover, preliminary clinical studies have been conducted and suggested PEMFs as a possible alternative therapy to treat acute ischemic stroke. In this work, we show that it's possible to build-up a patient semi-specific head model, where the 3D reconstruction of the ischemic lesion of the patient under treatment is inserted in the head of the human body model "Duke" (v.1.0, Zurich MedTech AG). The semi-specific model will be used in the randomized, placebo-controlled, double-blind study currently ongoing. Three patients were modelled and simulated, and results showed that each ischemic lesion experiences a magnetic flux density field comparable to the one for which biological effects have been attested. Such a kind of dosimetric analysis reveals a reliable tool to assess the correlation between levels of exposure and the beneficial effect. Thus, once the on-going double blind study is complete it will prove if PEMFs treatment triggers a clinical effect, and we will then be able to characterize a dose-response curve with the methodology arranged in this study.

Wireless epidermal devices based on Radio frequency Identification (RFID) enable a contactless and non-invasive monitoring of the human body by sampling health parameters directly on the skin. To achieve multi-parametric sensing, while preserving the intrinsic simplicity and the low cost of RFID tags, a dual-chip epidermal device is here proposed. At this purpose a polarization-diversity loop antenna is exploited so that two almost independent current modes are excited. The resulting radiation patterns are both broadside, thus enabling the simultaneous gathering of two independent dataset from the same maximum distance. A 3.5 by 3.5 cm battery-less, flexible and soft prototype provides -13 dBi embedded realized gain with read distances ranging from 0.6m to 1.5m depending on the microchip sensitivity. The electromagnetic performance of the two ports remain similar even when the tag is applied onto rather in-homogeneous body regions. With reference to body temperature monitoring, the device has been experimented in both controlled and real-life environments, demonstrating the possibility of doubling the sensing capabilities of RFID epidermal devices without affecting their size and radiation performances.

A proper choice of the scintillation crystal can considerably improve the detector performance in nuclear medicine imaging. In this study two promising crystals, CRY018 and CRY019, are characterized in terms of spectrometric and imaging capabilities. Thanks to the same irradiation conditions, it has been possible to compare their performance for a wide range of energies (30 keV to 662 keV) . They have high energy resolution (6.5% and 7.6% at 662 keV for CRY018 and CRY019, respectively), negligible light yield non-proportionality, good position linearity, and intrinsic spatial resolution less than 2 mm. In particular, the performance deviations could be explained from the different attenuation length (CRY018 depth of interaction is higher than CRY019 one, due to their different density).