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Abstract Electrical pulse stimulation has been used to enhance the differentiation or proliferation of neuronal progenitor cells in tissue engineering and cancer treatment. Therefore, a comprehensive investigation of the effects caused by its parameters is crucial for improvements in those fields. We propose a study of pulse parameters, to allow the control of N2a cell line fate and behavior. We have focused on designing an experimental setup that allows for the knowledge and control over the environment and the stimulation signals applied. To map the effects of the stimulation on N2a cells, their morphology and the cellular and molecular reactions induced by the pulse stimulation have been analyzed. Immunofluorescence, rt-PCR and western blot analysis have been carried out for this purpose, as well as cell counting. Our results show that low-amplitude electrical pulse stimulation promotes proliferation of N2a cells, whilst amplitudes in the range 250 mV/mm–500 mV/mm induce differentiation. Amplitudes higher than 750 mV/mm produce cell damage at low frequencies. For high frequencies, large amplitudes are needed to cause cell death. An inverse relation has been found between cell density and pulse-induced neuronal differentiation. The best condition for neuronal differentiation was found to be 500 mV/mm at 100 Hz. These findings have been confirmed by up-regulation of the Neurod1 gene. Our preliminary study of the molecular effects of electrical pulse stimulation on N2a offers premonitory clues of the PI3K/Akt/GSK-3β pathway implications on the neuronal differentiation process through ES. In general, we have successfully mapped the sensitivity of N2a cells to electrical pulse stimulation parameters.

Heart-failure (HF) is a severe medical condition. Physicians need new tools to monitor the health status of their HF patients outside the hospital or medical supervision areas, to better know the evolution of their patients’ main biomarker values, necessary to evaluate their health status. Bioimpedance (BI) represents a good technology for sensing physiological variables and processes on the human body. BI is a non-expensive and non-invasive technique for sensing a wide variety of physiological parameters, easy to be implemented on biomedical portable systems, also called “wearable devices”. In this systematic review, we address the most important specifications of wearable devices based on BI used in HF real-time monitoring and how they must be designed and implemented from a practical and medical point of view. The following areas will be analyzed: the main applications of BI in heart failure, the sensing technique and impedance specifications to be met, the electrode selection, portability of wearable devices: size and weight (and comfort), the communication requests and the power consumption (autonomy). The different approaches followed by biomedical engineering and clinical teams at bibliography will be described and summarized in the paper, together with results derived from the projects and the main challenges found today.

Background: Heart failure (HF) represents a frequent cause of hospital admission, with fluid overload directly contributing to decompensations. Bioimpedance (BI), a physical parameter linked to tissue hydration status, holds promise in monitoring congestion and improving prognosis. This systematic review aimed to assess the clinical relevance of BI-based wearable devices for HF fluid monitoring. Methods: A systematic review of the published literature was conducted in five medical databases (PubMed, Scopus, Cochrane, Web of Science, and Embase) for studies assessing wearable BI-measuring devices on HF patients following PRISMA recommendations on February 4th, 2024. The risk of bias was evaluated using the ROBINS tool. Results: The review included 10 articles with 535 participants (mean age 66.7 ± 8.9 years, males 70.4%). Three articles identified significant BI value differences between HF patients and controls or congestive vs non-congestive HF patients. Four articles focused on the devices' ability to predict HF worsening-related events, revealing an overall sensitivity of 70.0 (95% CI 68.8–71.1) and specificity of 89.1 (95% CI 88.3–89.9). One article assessed prognosis, showing that R80kHz decrease was related to all-cause-mortality with a hazard ratio (HR) of 5.51 (95% CI 1.55–23.32; p = 0.02) and the composite all-cause-mortality and HF admission with a HR of 4.96 (95% CI 1.82–14.37; p = 0.01). Conclusions: BI-measuring wearable devices exhibit efficacy in detecting fluid overload and hold promise for HF monitoring. However, further studies and technological improvements are required to optimize their impact on prognosis compared to standard care before they can be routinely implemented in clinical practice. PROSPERO Registration: The search protocol was registered at PROSPERO (CRD42024509914).

This paper provides an update on the sensors and actuators involved in Stem Cells (SC) differentiation processes based on electric stimulation (STIM), including both current and future progress. These techniques are applied in a range of biological and medical protocols, including cell linage derivation, tissue engineering, cellular therapy, cancer research, and cell motility. The typical methodology of SC electric STIM endeavors to emulate biological processes by applying an electrical signal to the cell culture and evaluating the cell response. Cell metabolism is electrically sensitive, responding in some manner to a given stimulus. The precise mechanism by which this occurs is not fully understood, but it is evident that changes in ion density at the cell membrane proximity must excite the cell metabolism (receptors), thereby activating its “differentiation” in response. In order to gain a deeper insight into the cellular mechanisms involved in this process, the physical variables should be better recognized, measured, and quantified during the protocol execution. This work is contributing to the development of a compilation of proposed systems, and specifications required, to identify and better understand the local conditions within the cell environment that are responsible for the activation of the differentiation processes. It is crucial that STIM systems are optimally designed and that the cell response is correctly understood. Two features will be reviewed: the setup employed and the circuits for STIM and monitoring. The nexus between these two elements are the electrodes, and this work will therefore be devoted to the realization of integrated Micro-Electrode Arrays (MEAs), and the design problems associated with it. The focus will be on MEAs, with the same size scale as the cells, and the design issues related to integrated electrodes, under electric stimulation, voltage or current modes.

Abstract Background Electrical stimulation is a novel tool to promote the differentiation and proliferation of precursor cells. In this work we have studied the effects of direct current (DC) electrical stimulation on neuroblastoma (N2a) and osteoblast (MC3T3) cell lines as a model for nervous and bone tissue regeneration, respectively. We have developed the electronics and encapsulation of a proposed stimulation system and designed a setup and protocol to stimulate cell cultures. Methods Cell cultures were subjected to several assays to assess the effects of electrical stimulation on them. N2a cells were analyzed using microscope images and an inmunofluorescence assay, differentiated cells were counted and neurites were measured. MC3T3 cells were subjected to an AlamarBlue assay for viability, ALP activity was measured, and a real time PCR was carried out. Results Our results show that electrically stimulated cells had more tendency to differentiate in both cell lines when compared to non-stimulated cultures, paired with a promotion of neurite growth and polarization in N2a cells and an increase in proliferation in MC3T3 cell line. Conclusions These results prove the effectiveness of electrical stimulation as a tool for tissue engineering and regenerative medicine, both for neural and bone injuries. Bone progenitor cells submitted to electrical stimulation have a higher tendency to differentiate and proliferate, filling the gaps present in injuries. On the other hand, neuronal progenitor cells differentiate, and their neurites can be polarized to follow the electric field applied.

Obtaining cell concentration measurements from a culture assay by using bioimpedance is a very useful method that can be used to translate impedances to cell concentration values. The purpose of this study was to find a method to obtain the cell concentration values of a given cell culture assay in real time by using an oscillator as the measurement circuit. From a basic cell–electrode model, enhanced models of a cell culture immersed in a saline solution (culture medium) were derived. These models were used as part of a fitting routine to estimate the cell concentration in a cell culture in real time by using the oscillation frequency and amplitude delivered by the measurement circuits proposed by previous authors. Using real experimental data (the frequency and amplitude of oscillations) that were obtained by connecting the cell culture to an oscillator as the load, the fitting routine was simulated, and real-time data of the cell concentration were obtained. These results were compared to concentration data that were obtained by using traditional optical methods for counting. In addition, the error that we obtained was divided and analyzed in two parts: the first part of the experiment (when the few cells were adapting to the culture medium) and the second part of the experiment (when the cells exponentially grew until they completely covered the well). Low error values were obtained during the growth phase of the cell culture (the relevant phase); therefore, the results obtained were considered promising and show that the fitting routine is valid and that the cell concentration can be measured in real time by using an oscillator.

Biological Impedance is a physical property related to the state and inherent evolution of biological samples. Among the existing impedance measurement methods, Oscillation-Based (OB) tests are a simple and smart solution to indirectly measure impedance correlated with the amplitude and frequency of the generated oscillation which are proportional to the sample under test. An OB test requires tuning of the system blocks to specifications derived from every measurement problem. The OB setup must be done to obtain the optimum measurement sensitivity for the specific constraints imposed by the system under test, electronic interfaces, and electrodes employed for test. This work proposes the extension of OB measurement systems to spectroscopy test, enabling a completely new range of applications for this technology without the restrictions imposed by setting a fixed frequency on the electrical oscillator. Some examples will be presented to the measurement of cell cultures samples, considering the corresponding circuit interfaces and electric models for the electrode-cell system. The proposed analysis method allows the selection of the best oscillator elements for optimum sensitivity range in amplitude and frequency oscillation values, when a specific cell culture is monitored for the OB system.

Coronary artery disease is one of the most common diseases in developed countries and affects a large part of the population of developing countries. Preventing restenosis in patients with implanted stents is an important current medical problem. The purpose of this work is to analyse the viability of bioimpedance sensing to detect the formation of atheromatous plaque in an implantable stent. Simulations in COMSOL Multiphysics were performed to analyse the performance of the proposed bioimpedance sensing system, based on the Sheffield technique. Both non-pathological and pathological models (with atheromatous plaque), including the flow of blood were considered. Simulations with the non-pathological model showed a homogeneous distribution of the measured current intensity in the different electrodes, for every configuration. On the other hand, simulations with the pathological model showed a significant decrease of the measured current intensity in the electrodes close to the simulated atheromatous plaque. The presence of the atheromatous plaque can, therefore, be detected by the system with a simple algorithm, avoiding the full reconstruction of the image and the subsequent computational processing requirements.

High-throughput data analysis challenges in laboratory automation and lab-on-a-chip devices’ applications are continuously increasing. In cell culture monitoring, specifically, the electrical cell-substrate impedance sensing technique (ECIS), has been extensively used for a wide variety of applications. One of the main drawbacks of ECIS is the need for implementing complex electrical models to decode the electrical performance of the full system composed by the electrodes, medium, and cells. In this work we present a new approach for the analysis of data and the prediction of a specific biological parameter, the fill-factor of a cell culture, based on a polynomial regression, data-analytic model. The method was successfully applied to a specific ECIS circuit and two different cell cultures, N2A (a mouse neuroblastoma cell line) and myoblasts. The data-analytic modeling approach can be used in the decoding of electrical impedance measurements of different cell lines, provided a representative volume of data from the cell culture growth is available, sorting out the difficulties traditionally found in the implementation of electrical models. This can be of particular importance for the design of control algorithms for cell cultures in tissue engineering protocols, and labs-on-a-chip and wearable devices applications.

A smart sensor system for cell culture real-time supervision is proposed, allowing for a significant reduction in human effort applied to this type of assay. The approach converts the cell culture under test into a suitable “biological” oscillator. The system enables the remote acquisition and management of the “biological” oscillation signals through a secure web interface. The indirectly observed biological properties are cell growth and cell number, which are straightforwardly related to the measured bio-oscillation signal parameters, i.e., frequency and amplitude. The sensor extracts the information without complex circuitry for acquisition and measurement, taking advantage of the microcontroller features. A discrete prototype for sensing and remote monitoring is presented along with the experimental results obtained from the performed measurements, achieving the expected performance and outcomes.

This paper proposes a new yet efficient method allowing a significant improvement in the on-line analysis of biological cell growing and evolution. The procedure is based on an empirical-mathematical approach for calibration and fitting of any cell-electrode electrical model. It is valid and can be extrapolated for any type of cellular line used in electrical cell-substrate impedance spectroscopy (ECIS) tests. Parameters of the bioimpedance model, acquired from ECIS experiments, vary for each cell line, which makes obtaining results difficult and—to some extent-renders them inaccurate. We propose a fitting method based on the cell line initial characterization, and carry out subsequent experiments with the same line to approach the percentage of well filling and the cell density (or cell number in the well). To perform our calibration technique, the so-called oscillation-based test (OBT) approach is employed for each cell density. Calibration results are validated by performing other experiments with different concentrations on the same cell line with the same measurement technique. Accordingly, a bioimpedance electrical model of each cell line is determined, which is valid for any further experiment and leading to a more precise electrical model of the electrode-cell system. Furthermore, the model parameters calculated can be also used by any other measurement techniques. Promising experimental outcomes for three different cell-lines have been achieved, supporting the usefulness of this technique.

To follow up the restenosis in arteries stented during an angioplasty is an important current clinical problem. A new approach to monitor the growth of neointimal tissue inside the stent is proposed on the basis of electrical impedance spectroscopy (EIS) sensors and the oscillation-based test (OBT) circuit technique. A mathematical model was developed to analytically describe the histological composition of the neointima, employing its conductivity and permittivity data. The bioimpedance model was validated against a finite element analysis (FEA) using COMSOL Multiphysics software. A satisfactory correlation between the analytical model and FEA simulation was achieved in most cases, detecting some deviations introduced by the thin “double layer” that separates the neointima and the blood. It is hereby shown how to apply conformal transformations to obtain bioimpedance electrical models for stack-layered tissues over coplanar electrodes. Particularly, this can be applied to characterize the neointima in real-time. This technique is either suitable as a main mechanism for restenosis follow-up or it can be combined with proposed intelligent stents for blood pressure measurements to auto-calibrate the sensibility loss caused by the adherence of the tissue on the micro-electro-mechanical sensors (MEMSs).

This paper presents the design of an optical Position Sensing Detector (PSD) for application in biological material gene identification. The system is able to measure changes in the position of a light spot emitted by a Vertical Cavity Surface Emitting Laser (VCSEL). Changes in spot position are produced by the deflections of a microcantilever used to sense the hybridisation process of the biological material. Detection of these deflections requires submicrometer resolution. The photodetectors and the detection algorithm proposed in this paper have been designed and optimized for this resolution. A 0.35 μm CMOS prototype consisting of an array of 22 PSDs has been implemented and experimentally characterized. The obtained results confirm that displacements of the light spot position as low as 0.15 μm can be detected. [ABSTRACT FROM AUTHOR]

In tissue engineering, of utmost importance is the control of tissue formation, in order to form tissue constructs of clinical relevance. In this work, we present the use of an impedance spectroscopy technique for the real-time measurement of the dielectric properties of skeletal myoblast cell cultures. The processes involved in the growth and differentiation of these cell cultures in skeletal muscle are studied. A circuit based on the oscillation-based test technique was used, avoiding the use of high-performance circuitry or external input signals. The effect of electrical pulse stimulation applied to cell cultures was also studied. The technique proved useful for monitoring in real-time the processes of cell growth and estimating the fill factor of muscular stem cells. Impedance spectroscopy was also useful to study the real-time monitoring of cell differentiation, obtaining different oscillation amplitude levels for differentiated and undifferentiated cell cultures. Finally, an electrical model was implemented to better understand the physical properties of the cell culture and control the tissue formation process.