Optical mapping and optogenetics in cardiac electrophysiology research and therapy:a state-of-the-art review

State-of-the-art innovations in optical cardiac electrophysiology are significantly enhancing cardiac research. A potential leap into patient care is now on the horizon. Optical mapping, using fluorescent probes and high-speed cameras, offers detailed insights into cardiac activity and arrhythmias by analysing electrical signals, calcium dynamics, and metabolism. Optogenetics utilizes light-sensitive ion channels and pumps to realize contactless, cell-selective cardiac actuation for modelling arrhythmia, restoring sinus rhythm, and probing complex cell–cell interactions. The merging of optogenetics and optical mapping techniques for ‘all-optical’ electrophysiology marks a significant step forward. This combination allows for the contactless actuation and sensing of cardiac electrophysiology, offering unprecedented spatial–temporal resolution and control. Recent studies have performed all-optical imaging ex vivo and achieved reliable optogenetic pacing in vivo, narrowing the gap for clinical use. Progress in optical electrophysiology continues at pace. Advances in motion tracking methods are removing the necessity of motion uncoupling, a key limitation of optical mapping. Innovations in optoelectronics, including miniaturized, biocompatible illumination and circuitry, are enabling the creation of implantable cardiac pacemakers and defibrillators with optoelectrical closed-loop systems. Computational modelling and machine learning are emerging as pivotal tools in enhancing optical techniques, offering new avenues for analysing complex data and optimizing therapeutic strategies. However, key challenges remain including opsin delivery, real-time data processing, longevity, and chronic effects of optoelectronic devices. This review provides a comprehensive overview of recent advances in optical mapping and optogenetics and outlines the promising future of optics in reshaping cardiac electrophysiology and therapeutic strategies

Digital neural circuits : from ions to networks

PhD ThesisThe biological neural computational mechanism is always fascinating to human beings since it shows several state-of-the-art characteristics: strong fault tolerance, high power efficiency and self-learning capability. These behaviours lead the developing trend of designing the next-generation digital computation platform. Thus investigating and understanding how the neurons talk with each other is the key to replicating these calculation features. In this work I emphasize using tailor-designed digital circuits for exactly implementing bio-realistic neural network behaviours, which can be considered a novel approach to cognitive neural computation. The first advance is that biological real-time computing performances allow the presented circuits to be readily adapted for real-time closed-loop in vitro or in vivo experiments, and the second one is a transistor-based circuit that can be directly translated into an impalpable chip for high-level neurologic disorder rehabilitations. In terms of the methodology, first I focus on designing a heterogeneous or multiple-layer-based architecture for reproducing the finest neuron activities both in voltage-and calcium-dependent ion channels. In particular, a digital optoelectronic neuron is developed as a case study. Second, I focus on designing a network-on-chip architecture for implementing a very large-scale neural network (e.g. more than 100,000) with human cognitive functions (e.g. timing control mechanism). Finally, I present a reliable hybrid bio-silicon closed-loop system for central pattern generator prosthetics, which can be considered as a framework for digital neural circuit-based neuro-prosthesis implications. At the end, I present the general digital neural circuit design principles and the long-term social impacts of the presented work

Optogenetics in striated muscle: defibrillation of the heart and direct stimulation of skeletal muscles with light

Optogenetic depolarization of cells using the non-selective cation channel Channelrhodopsin-2 (ChR2) enables precise control over the membrane potential of cells within a specific area of intact organs. Furthermore, the selective overexpression of light-gated proteins allows cell type-specific and pain-free stimulation which could be of great benefit for future scientific and therapeutic approaches. In my thesis, I explored two potential applications of optogenetic methods in striated muscle: optogenetic defibrillation to terminate ventricular arrhythmia in intact mouse hearts and direct optogenetic stimulation of skeletal muscles. These new approaches could lead in the future to the development of optogenetic defibrillators and laryngeal pacemakers. Most experiments were performed with explanted hearts, isolated skeletal fibers and muscles or larynges from transgenic ChR2 expressing mice. To add translational perspectives, we also explored optogenetic defibrillation and intralaryngeal muscles stimulation after ChR2 gene transfer to wild type mice using adeno-associated virus (AAV). Optogenetic defibrillation by epicardial illumination was highly efficient in terminating ventricular arrhythmia in transgenic hearts and the success rate of optogenetic defibrillation was depending on the pulse duration, the size of illumination and the light intensity. Importantly, we were also able to terminate ventricular arrhythmia in non-transgenic hearts even one year after AAV mediated gene transfer. The potential applicability of optogenetic defibrillation in the human heart was assessed in experimentally-calibrated computer simulations of a patient’s heart with infarct-related ventricular tachycardia. Because optogenetic stimulation would be in principle pain-free in patients, the proof for its feasibility could lay the foundation for the development of a new treatment option for patients at high risk for ventricular arrhythmia. Direct optogenetic stimulation of skeletal muscle was first proven in isolated Flexor digitorum brevis fibers and in intact soleus muscles, which could both be stimulated using brief light pulses. The force of light-induced single twitches could be precisely controlled by varying the pulse duration and light intensity. Optogenetic stimulation was most efficient with 10 ms long pulses at a repetition rate of 40 Hz reaching ~84% of the maximum force generated by electrical stimulation with 100 Hz. Recurrent nerve paralysis is a severe complication of neck surgery, malignant processes or central neurological diseases and results in a fixed paramedian position of the vocal cords as well as life-threatening dyspnea in the case of bilateral paralysis. Current treatment options consist only of destructive surgery. Unfortunately the use of local electrical stimulation to restore laryngeal function faces severe technical limitations. Therefore I sought to explore direct optogenetic stimulation of intralaryngeal muscles in explanted larynges from ChR2 transgenic mice. Specific illumination of the individual intralaryngeal muscle groups led to an opening or closing of the vocal cords depending on the site of illumination. This proves the sufficient spatial resolution of light for selective stimulation of the intralaryngeal muscles groups. In addition, we were able to induce opening of the vocal cords in wild type mice after AAV-based gene transfer of ChR2 with light. Thus optogenetic stimulation could become a new treatment option for patients suffering from bilateral laryngeal paralysis. In conclusion, optogenetic stimulation can overcome the severe limitations of electrical stimulation of the heart and skeletal muscles. The new technologies, I have developed and characterized in this thesis, allow for the design of completely new stimulation patterns to address open questions in muscle physiology. Furthermore, optogenetic stimulation of striated muscles could become a new treatment option for patients enabling selective and pain-free stimulation with few side effects

Energy Conversion from Gradients across Bio-Inspired Membranes

Membranes are fundamentally important barriers that enable the processes of life by slowing the dissipation of gradients and transducing usable energy from entropic driving forces. This dissertation presents three investigations of gradient equilibration across bio-inspired membranes, with a focus on permeability and geometric considerations in membrane-based energy transduction systems. The first study models the dynamics of pressure generation from osmotic gradients in an expandable compartment based on mass transport principles. Using an osmotic working fluid composed of aqueous polyethylene glycol inside commercially available dialysis cassettes whose membranes exclude polymer solutes, we validated this model and explored the importance of cassette geometry, restrictions on expansion, and membrane porosity characteristics on pressure generation rates over time. The model made it possible to predict the kinetics of nastic motions caused by osmotically-induced shifts in turgor pressure in plants such as Mimosa pudica; the model’s projections based on plant cell dimensions agree well with published time scales. These cassettes are “waterable” pressure generators available to the general public; we demonstrated their utility by actuating a soft robotic gripper and published our characterization algorithm as an open-source tool. The second study investigates the relationship between the chemical structures of a class of tethered membrane-spanning lipids found in hyperthermoacidophilic archaea and the proton/hydroxide permeability of self-assembled monolayer membranes that the lipids form. We determined permeability values by measuring the fluorescence of solutions of liposomes containing pH-sensitive dyes over time after a step change in the external pH. The presence of isoprenoid methyl groups led to reductions in permeability values, and the length of the transmembrane tether unexpectedly displayed a direct correlation with the permeability, leading us to speculate about the importance of hydrophobic crowding in the membrane interior. Surprisingly, the presence of a transmembrane tether had no significant effect on the permeability at room temperature. We observed a strong positive correlation between the permeability of a membrane and the probability of observing water molecules spontaneously clustering inside the hydrophobic region of a simulated membrane of identical composition using molecular dynamics, providing a predictive parameter obtainable without any “wet” experimentation that may be useful for the design of membrane compositions with specific permeability characteristics. The third study presents a novel hydrogel-based reverse-electrodialytic energy transduction scheme inspired by the electric eel. As in the biological system (but unlike traditional batteries), the “artificial electric organ” presented here is a soft, flexible, potentially biocompatible means of generating electricity using only a repeating arrangement of ionic gradients across selective membranes. The artificial electric organ generates numerous additive voltages at the same time in order to produce a large transient electric signal; while the biological system accomplishes this synchronicity through neural signaling, the artificial setup uses geometries that allow simultaneous mechanical registration of the gels. This scheme is flexible enough that we were able to implement it in three distinct ways: fluidically, through the printing of large arrays (enabling the generation of over 100 V), and using a Miura-ori folding geometry that assembles planar films, which achieved a power density of 27 mW m^-2. The work presented here draws inspiration from biological systems and earlier industrial efforts to extract electrical power from salinity gradients. Membrane-based strategies are well-suited for the local generation of usable energy on small scales and may be useful in the microelectromechanical systems and implantable devices of the future.PHDChemical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttps://deepblue.lib.umich.edu/bitstream/2027.42/140979/1/tomschro_1.pd

Advances in Bioengineering

The technological approach and the high level of innovation make bioengineering extremely dynamic and this forces researchers to continuous updating. It involves the publication of the results of the latest scientific research. This book covers a wide range of aspects and issues related to advances in bioengineering research with a particular focus on innovative technologies and applications. The book consists of 13 scientific contributions divided in four sections: Materials Science; Biosensors. Electronics and Telemetry; Light Therapy; Computing and Analysis Techniques

Advance in Energy Harvesters/Nanogenerators and Self-Powered Sensors

This reprint is a collection of the Special Issue "Advance in Energy Harvesters/Nanogenerators and Self-Powered Sensors" published in Nanomaterials, which includes one editorial, six novel research articles and four review articles, showcasing the very recent advances in energy-harvesting and self-powered sensing technologies. With its broad coverage of innovations in transducing/sensing mechanisms, material and structural designs, system integration and applications, as well as the timely reviews of the progress in energy harvesting and self-powered sensing technologies, this reprint could give readers an excellent overview of the challenges, opportunities, advancements and development trends of this rapidly evolving field

Intelligent Biosignal Processing in Wearable and Implantable Sensors

This reprint provides a collection of papers illustrating the state-of-the-art of smart processing of data coming from wearable, implantable or portable sensors. Each paper presents the design, databases used, methodological background, obtained results, and their interpretation for biomedical applications. Revealing examples are brain–machine interfaces for medical rehabilitation, the evaluation of sympathetic nerve activity, a novel automated diagnostic tool based on ECG data to diagnose COVID-19, machine learning-based hypertension risk assessment by means of photoplethysmography and electrocardiography signals, Parkinsonian gait assessment using machine learning tools, thorough analysis of compressive sensing of ECG signals, development of a nanotechnology application for decoding vagus-nerve activity, detection of liver dysfunction using a wearable electronic nose system, prosthetic hand control using surface electromyography, epileptic seizure detection using a CNN, and premature ventricular contraction detection using deep metric learning. Thus, this reprint presents significant clinical applications as well as valuable new research issues, providing current illustrations of this new field of research by addressing the promises, challenges, and hurdles associated with the synergy of biosignal processing and AI through 16 different pertinent studies. Covering a wide range of research and application areas, this book is an excellent resource for researchers, physicians, academics, and PhD or master students working on (bio)signal and image processing, AI, biomaterials, biomechanics, and biotechnology with applications in medicine

New Techniques in Gastrointestinal Endoscopy

As result of progress, endoscopy has became more complex, using more sophisticated devices and has claimed a special form. In this moment, the gastroenterologist performing endoscopy has to be an expert in macroscopic view of the lesions in the gut, with good skills for using standard endoscopes, with good experience in ultrasound (for performing endoscopic ultrasound), with pathology experience for confocal examination. It is compulsory to get experience and to have patience and attention for the follow-up of thousands of images transmitted during capsule endoscopy or to have knowledge in physics necessary for autofluorescence imaging endoscopy. Therefore, the idea of an endoscopist has changed. Examinations mentioned need a special formation, a superior level of instruction, accessible to those who have already gained enough experience in basic diagnostic endoscopy. This is the reason for what these new issues of endoscopy are presented in this book of New techniques in Gastrointestinal Endoscopy

Multimodal Investigation of the Efficiency and Stability of Microstimulation using Electrodes Coated with PEDOT/CNT and Iridium Oxide

Electrical microstimulation is an invaluable tool in neuroscience research to dissect neural circuits, relate brain areas, and identify relationships between brain structure and behavior. In the clinic, electrical microstimulation has enabled partial restoration of vision, movement, sensation and autonomic functions. Recently, novel materials and new fabrication techniques of traditional metals have emerged such as iridium oxide and the conducting polymer PEDOT/CNT. These materials have demonstrated particular promise in the improvement in electrical efficiency. However, the in vivo stimulation efficiency and the in vivo stability of these materials have not been thoroughly characterized. In this dissertation, we use a multimodal approach to study the efficiency and stability of electrode-tissue interface using novel materials in microstimulation

Ion camera development for real–time acquisition of localised pH responses using the CMOS based 64×64–pixel ISFET sensor array technology

This thesis presents the development and test of an integrated ion camera chip for monitoring highly localised ion fluxes of electrochemical processes using an ion sensitive sensor array. Ionic concentration fluctuations are shown to travel across the sensor array as a result of citric acid injection and the BZ-reaction. The imaging capability of non-equilibrium chemical activities is also demonstrated monitoring self-assembling micrometre sized polyoxometalate tubular and membranous architectures. The sufficient spatial resolution for the visualisation of the 10-60 µm wide growing trajectories is provided by the dense sensor array containing 64×64 pixels. In the case of citric acid injection and the BZ-reaction the ion camera chip is shown to be able to resolve pH differences with resolution as low as the area of one pixel. As a result of the transient and volatile ionic fluxes high time resolution is required, thus the signal capturing can be performed in real.time at the maximum sampling rate of 40 µs per pixel, 10.2 ms per array. The extracted sensor data are reconstructed into ionic images and thus the ionic activities can be displayed as individual figures as well as continuous video recordings. This chip is the first prototype in the envisioned establishment of a fully automated CMOS based ion camera system which would be able to image the invisible activity of ions using a single microchip. In addition the capability of detecting ultra-low level pH oscillations in the extracellular space is demonstrated using cells of the slime mould organism. The detected pH oscillations with extent of ~0.022 pH furthermore raise the potential for observing fluctuations of ion currents in cell based tissue environments. The intrinsic noise of the sensor devices are measured to observe noise effect on the detected low level signals. It is experimentally shown that the used ion sensitive circuits, similarly to CMOS, also demonstrate 1/f noise. In addition the reference bias and pH sensitivity of the measured noise is confirmed. Corresponding to the measurement results the noise contribution is approximated with a 28.2 µV peak-to-peak level and related to the 450 µV �+/- 70 µV peak-to-peak oscillations amplitudes of the slime mould. Thus a maximum intrinsic noise contribution of 6.2 �+/- 1.2 % is calculated. A H+ flickering hypothesis is also presented that correlates the pH fluctuations on the surface of the device with the intrinsic 1/f noise. The ion camera chip was fabricated in an unmodified 4-metal 0.35 µm CMOS process and the ionic imaging technology was based on a 64�×64-pixel ion sensitive field effect transistor (ISFET) array. The high-speed and synchronous operation of the 4096 ISFET sensors occupying 715.8×715.8 µm space provided a spatial resolution as low as one pixel. Each pixel contained 4 transistors with 10.2×10.2 µm layout dimensions and the pixels were separated by a 1 µm separation gap. The ion sensitive silicon nitride based passivation layer was in contact with the floating gates of the ISFET sensors. It allowed the capacitive measurements of localised changes in the ionic concentrations, e.g. pH, pNa, on the surface of the chip. The device showed an average ionic sensitivity of 20 mV/pH and 9 mV/pNa. The packaging and encapsulation was carried out using PGA-100 chip carriers and two-component epoxies. Custom designed printed circuit boards (PCBs) were used to provide interface between the ISFET array chip and the data acquisition system. The data acquisition and extraction part of the developed software system was based on LabVIEW, the data processing was carried out on Matlab platform