A soft, scalable and adaptable multi-contact cuff electrode for targeted peripheral nerve modulation.

BACKGROUND: Cuff electrodes target various nerves throughout the body, providing neuromodulation therapies for motor, sensory, or autonomic disorders. However, when using standard, thick silicone cuffs, fabricated in discrete circular sizes, complications may arise, namely cuff displacement or nerve compression, due to a poor adaptability to variable nerve shapes and sizes encountered in vivo. Improvements in cuff design, materials, closing mechanism and surgical approach are necessary to overcome these issues. METHODS: In this work, we propose a microfabricated multi-channel silicone-based soft cuff electrode with a novel easy-to-implant and size-adaptable design and evaluate a number of essential features such as nerve-cuff contact, nerve compression, cuff locking stability, long-term integration and stimulation selectivity. We also compared performance to that of standard fixed-size cuffs. RESULTS: The belt-like cuff made of 150 µm thick silicone membranes provides a stable and pressure-free conformal contact, independently of nerve size variability, combined with a straightforward implantation procedure. The adaptable design and use of soft materials lead to limited scarring and demyelination after 6-week implantation. In addition, multi-contact designs, ranging from 6 to 16 electrodes, allow for selective stimulation in models of rat and pig sciatic nerve, achieving targeted activation of up to 5 hindlimb muscles. CONCLUSION: These results suggest a promising alternative to classic fixed-diameter cuffs and may facilitate the adoption of soft, adaptable cuffs in clinical settings.

Optical Monitoring of Water Side Permeation in Thin Film Encapsulation.

The stability of long-term microfabricated implants is hindered by the presence of multiple water diffusion paths within artificially patterned thin-film encapsulations. Side permeation, defined as infiltration of molecules through the lateral surface of the thin structure, becomes increasingly critical with the trend of developing high-density and miniaturized neural electrodes. However, current permeability measurement methods do not account for side permeation accurately nor quantitatively. Here, a novel optical, magnesium (Mg)-based method is proposed to quantify the side water transmission rate (SWTR) through thin film encapsulation and validate the approach using micrometric polyimide (PI) and polyimide-silicon carbide (PI-SiC) multilayers. Through computed digital grayscale images collected with corroding Mg film microcells coated with the thin encapsulation, side and surface WTRs are quantified. A 4.5-fold ratio between side and surface permeation is observed, highlighting the crucial role of the PI-PI interface in lateral diffusion. Universal guidelines for the design of flexible, hermetic neural interfaces are proposed. Increasing encapsulation's width (interelectrode spacing), creating stronger interfacial interactions, and integrating high-barrier interlayers such as SiC significantly enhance the lateral hermeticity.

Radiation Patterns of RF Wireless Devices Implanted in Small Animals: Unexpected Deformations Due to Body Resonance.

One challenge in designing RF wireless bioelectronic devices is the impact of the interaction between electromagnetic waves and host body tissues on far-field wireless performance. In this article, we investigate a peculiar phenomenon of implantable RF wireless devices within a small-scale host body related to the deformation of the directivity pattern. Radiation measurements of subcutaneously implanted antennas within rodent cadavers show that the direction of maximum radiation is not always identical with the direction to the closest body-air interface, as one would expect in larger-scale host bodies. For an implanted antenna in the back of a mouse, we observed the maximum directivity in the ventral direction with 4.6 dB greater gain compared to the nearest body-air interface direction. Analytic analysis within small-scale spherical body phantoms identifies two main factors for these results: the limited absorption losses due to the small body size relative to the operating wavelength and the high permittivity of the biological tissues of the host body. Due to these effects, the entire body acts as a dielectric resonator antenna, leading to deformations of the directivity pattern. These results are confirmed with the practical example of a wirelessly powered 2.4-GHz optogenetic implant, demonstrating the significance of the judicious placement of external antennas to take advantage of the deformation of the implanted antenna pattern. These findings emphasize the importance of carefully designing implantable RF wireless devices based on their placements and relative electrical dimensions in small-scale animal models.

A low-profile electromechanical packaging system for soft-to-flexible bioelectronic interfaces.

Interfacing the human body with the next generation of electronics requires technological advancement in designing and producing bioelectronic circuits. These circuits must integrate electrical functionality while simultaneously addressing limitations in mechanical compliance and dynamics, biocompatibility, and consistent, scalable manufacturing. The combination of mechanically disparate materials ranging from elastomers to inorganic crystalline semiconductors calls for modular designs with reliable and scalable electromechanical connectors. Here, we report on a novel interconnection solution for soft-to-flexible bioelectronic interfaces using a patterned and machined flexible printed circuit board, which we term FlexComb, interfaced with soft transducing systems. Using a simple assembly process, arrays of protruding "fingers" bearing individual electrical terminals are laser-machined on a standard flexible printed circuit board to create a comb-like structure, namely, the FlexComb. A matching pattern is also machined in the soft system to host and interlock electromechanically the FlexComb connections via a soft electrically conducting composite. We examine the electrical and electromechanical properties of the interconnection and demonstrate the versatility and scalability of the method through various customized submillimetric designs. In a pilot in vivo study, we validate the stability and compatibility of the FlexComb technology in a subdural electrocorticography system implanted for 6 months on the auditory cortex of a minipig. The FlexComb provides a reliable and simple technique to bond and connect soft transducing systems with flexible or rigid electronic boards, which should find many implementations in soft robotics and wearable and implantable bioelectronics.

Deployment of an electrocorticography system with a soft robotic actuator.

Electrocorticography (ECoG) is a minimally invasive approach frequently used clinically to map epileptogenic regions of the brain and facilitate lesion resection surgery and increasingly explored in brain-machine interface applications. Current devices display limitations that require trade-offs among cortical surface coverage, spatial electrode resolution, aesthetic, and risk consequences and often limit the use of the mapping technology to the operating room. In this work, we report on a scalable technique for the fabrication of large-area soft robotic electrode arrays and their deployment on the cortex through a square-centimeter burr hole using a pressure-driven actuation mechanism called eversion. The deployable system consists of up to six prefolded soft legs, and it is placed subdurally on the cortex using an aqueous pressurized solution and secured to the pedestal on the rim of the small craniotomy. Each leg contains soft, microfabricated electrodes and strain sensors for real-time deployment monitoring. In a proof-of-concept acute surgery, a soft robotic electrode array was successfully deployed on the cortex of a minipig to record sensory cortical activity. This soft robotic neurotechnology opens promising avenues for minimally invasive cortical surgery and applications related to neurological disorders such as motor and sensory deficits.

Sensitivity to Pulse Rate and Amplitude Modulation in an Animal Model of the Auditory Brainstem Implant (ABI).

The auditory brainstem implant (ABI) is an auditory neuroprosthesis that provides hearing by electrically stimulating the cochlear nucleus (CN) of the brainstem. Our previous study (McInturff et al., 2022) showed that single-pulse stimulation of the dorsal (D)CN subdivision with low levels of current evokes responses that have early latencies, different than the late response patterns observed from stimulation of the ventral (V)CN. How these differing responses encode more complex stimuli, such as pulse trains and amplitude modulated (AM) pulses, has not been explored. Here, we compare responses to pulse train stimulation of the DCN and VCN, and show that VCN responses, measured in the inferior colliculus (IC), have less adaption, higher synchrony, and higher cross-correlation. However, with high-level DCN stimulation, responses become like those to VCN stimulation, supporting our earlier hypothesis that current spreads from electrodes on the DCN to excite neurons located in the VCN. To AM pulses, stimulation of the VCN elicits responses with larger vector strengths and gain values especially in the high-CF portion of the IC. Additional analysis using neural measures of modulation thresholds indicate that these measures are lowest for VCN. Human ABI users with low modulation thresholds, who score best on comprehension tests, may thus have electrode arrays that stimulate the VCN. Overall, the results show that the VCN has superior response characteristics and suggest that it should be the preferred target for ABI electrode arrays in humans.

Subdural Soft Electrocorticography (ECoG) Array Implantation and Long-Term Cortical Recording in Minipigs.

Neurological impairments and diseases can be diagnosed or treated using electrocorticography (ECoG) arrays. In drug-resistant epilepsy, these help delineate the epileptic region to resect. In long-term applications such as brain-computer interfaces, these epicortical electrodes are used to record the movement intention of the brain, to control the robotic limbs of paralyzed patients. However, current stiff electrode grids do not answer the need for high-resolution brain recordings and long-term biointegration. Recently, conformable electrode arrays have been proposed to achieve long-term implant stability with high performance. However, preclinical studies for these new implant technologies are needed to validate their long-term functionality and safety profile for their translation to human patients. In this context, porcine models are routinely employed in developing medical devices due to their large organ sizes and easy animal handling. However, only a few brain applications are described in the literature, mostly due to surgery limitations and integration of the implant system on a living animal. Here, we report the method for long-term implantation (6 months) and evaluation of soft ECoG arrays in the minipig model. The study first presents the implant system, consisting of a soft microfabricated electrode array integrated with a magnetic resonance imaging (MRI)-compatible polymeric transdermal port that houses instrumentation connectors for electrophysiology recordings. Then, the study describes the surgical procedure, from subdural implantation to animal recovery. We focus on the auditory cortex as an example target area where evoked potentials are induced by acoustic stimulation. We finally describe a data acquisition sequence that includes MRI of the whole brain, implant electrochemical characterization, intraoperative and freely moving electrophysiology, and immunohistochemistry staining of the extracted brains. This model can be used to investigate the safety and function of novel design of cortical prostheses; mandatory preclinical study to envision translation to human patients.

Cardiovascular Response to Intraneural Right Vagus Nerve Stimulation in Adult Minipig.

OBJECTIVE: This study explored intraneural stimulation of the right thoracic vagus nerve (VN) in sexually mature male minipigs to modulate safe heart rate and blood pressure response. MATERIAL AND METHODS: We employed an intraneural electrode designed for the VN of pigs to perform VN stimulation (VNS). This was delivered using different numbers of contacts on the electrode and different stimulation parameters (amplitude, frequency, and pulse width), identifying the most suitable stimulation configuration. All the parameter ranges had been selected from a computational cardiovascular system model. RESULTS: Clinically relevant responses were observed when stimulating with low current intensities and relatively low frequencies delivered with a single contact. Selecting a biphasic, charge-balanced square wave for VNS with a current amplitude of 500 µA, frequency of 10 Hz, and pulse width of 200 µs, we obtained heart rate reduction of 7.67 ± 5.19 beats per minute, systolic pressure reduction of 5.75 ± 2.59 mmHg, and diastolic pressure reduction of 3.39 ± 1.44 mmHg. CONCLUSION: Heart rate modulation was obtained without inducing any observable adverse effects, underlining the high selectivity of the intraneural approach.

Revealing the complexity of ultra-soft hydrogel re-swelling inside the brain.

The brain is an ultra-soft viscoelastic matrix. Sub-kPa hydrogels match the brain's mechanical properties but are challenging to manipulate in an implantable format. We propose a simple fabrication and processing sequence, consisting of de-hydration, patterning, implantation, and re-hydration steps, to deliver brain-like hydrogel implants into the nervous tissue. We monitored in real-time the ultra-soft hydrogel re-swelling kinetics in vivo using microcomputed tomography, achieved by embedding gold nanoparticles inside the hydrogel for contrast enhancement. We found that re-swelling in vivo strongly depends on the implant geometry and water availability at the hydrogel-tissue interface. Buckling of the implant inside the brain occurs when the soft implant is tethered to the cranium. Finite-element and analytical models reveal how the shank geometry, modulus and anchoring govern in vivo buckling. Taken together, these considerations on re-swelling kinetics of hydrogel constructs, implant geometry and soft implant-tissue mechanical interplay can guide the engineering of biomimetic brain implants.

The neurons that restore walking after paralysis.

A spinal cord injury interrupts pathways from the brain and brainstem that project to the lumbar spinal cord, leading to paralysis. Here we show that spatiotemporal epidural electrical stimulation (EES) of the lumbar spinal cord1-3 applied during neurorehabilitation4,5 (EESREHAB) restored walking in nine individuals with chronic spinal cord injury. This recovery involved a reduction in neuronal activity in the lumbar spinal cord of humans during walking. We hypothesized that this unexpected reduction reflects activity-dependent selection of specific neuronal subpopulations that become essential for a patient to walk after spinal cord injury. To identify these putative neurons, we modelled the technological and therapeutic features underlying EESREHAB in mice. We applied single-nucleus RNA sequencing6-9 and spatial transcriptomics10,11 to the spinal cords of these mice to chart a spatially resolved molecular atlas of recovery from paralysis. We then employed cell type12,13 and spatial prioritization to identify the neurons involved in the recovery of walking. A single population of excitatory interneurons nested within intermediate laminae emerged. Although these neurons are not required for walking before spinal cord injury, we demonstrate that they are essential for the recovery of walking with EES following spinal cord injury. Augmenting the activity of these neurons phenocopied the recovery of walking enabled by EESREHAB, whereas ablating them prevented the recovery of walking that occurs spontaneously after moderate spinal cord injury. We thus identified a recovery-organizing neuronal subpopulation that is necessary and sufficient to regain walking after paralysis. Moreover, our methodology establishes a framework for using molecular cartography to identify the neurons that produce complex behaviours.

A finite element model of the mechanical interactions between peripheral nerves and intrafascicular implants.

Objective.Intrafascicular peripheral nerve implants are key components in the development of bidirectional neuroprostheses such as touch-enabled bionic limbs for amputees. However, the durability of such interfaces is hindered by the immune response following the implantation. Among the causes linked to such reaction, the mechanical mismatch between host nerve and implant is thought to play a decisive role, especially in chronic settings.Approach.Here we focus on modeling mechanical stresses induced on the peripheral nerve by the implant's micromotion using finite element analysis. Through multiple parametric sweeps, we analyze the role of the implant's material, geometry (aspect-ratio and shape), and surface coating, deriving a set of parameters for the design of better-integrated implants.Main results.Our results indicate that peripheral nerve implants should be designed and manufactured with smooth edges, using materials at most three orders of magnitude stiffer than the nerve, and with innovative geometries to redistribute micromotion-associated loads to less delicate parts of the nerve such as the epineurium.Significance.Overall, our model is a useful tool for the peripheral nerve implant designer that is mindful of the importance of implant mechanics for long term applications.

Active force generation contributes to the complexity of spontaneous activity and to the response to stretch of murine cardiomyocyte cultures.

Cardiomyocyte cultures exhibit spontaneous electrical and contractile activity, as in a natural cardiac pacemaker. In such preparations, beat rate variability exhibits features similar to those of heart rate variability in vivo. Mechanical deformations and forces feed back on the electrical properties of cardiomyocytes, but it is not fully elucidated how this mechano-electrical interplay affects beating variability in such preparations. Using stretchable microelectrode arrays, we assessed the effects of the myosin inhibitor blebbistatin and the non-selective stretch-activated channel blocker streptomycin on beating variability and on the response of neonatal or fetal murine ventricular cell cultures against deformation. Spontaneous electrical activity was recorded without stretch and upon predefined deformation protocols (5% uniaxial and 2% equibiaxial strain, applied repeatedly for 1 min every 3 min). Without stretch, spontaneous activity originated from the edge of the preparations, and its site of origin switched frequently in a complex manner across the cultures. Blebbistatin did not change mean beat rate, but it decreased the spatial complexity of spontaneous activity. In contrast, streptomycin did not exert any manifest effects. During the deformation protocols, beat rate increased transiently upon stretch but, paradoxically, also upon release. Blebbistatin attenuated the response to stretch, whereas this response was not affected by streptomycin. Therefore, our data support the notion that in a spontaneously firing network of cardiomyocytes, active force generation, rather than stretch-activated channels, is involved mechanistically in the complexity of the spatiotemporal patterns of spontaneous activity and in the stretch-induced acceleration of beating. KEY POINTS: Monolayer cultures of cardiac cells exhibit spontaneous electrical and contractile activity, as in a natural cardiac pacemaker. Beating variability in these preparations recapitulates the power-law behaviour of heart rate variability in vivo. However, the effects of mechano-electrical feedback on beating variability are not yet fully understood. Using stretchable microelectrode arrays, we examined the effects of the contraction uncoupler blebbistatin and the non-specific stretch-activated channel blocker streptomycin on beating variability and on stretch-induced changes of beat rate. Without stretch, blebbistatin decreased the spatial complexity of beating variability, whereas streptomycin had no effects. Both stretch and release increased beat rate transiently; blebbistatin attenuated the increase of beat rate upon stretch, whereas streptomycin had no effects. Active force generation contributes to the complexity of spatiotemporal patterns of beating variability and to the increase of beat rate upon mechanical deformation. Our study contributes to the understanding of how mechano-electrical feedback influences heart rate variability.

Comparison of Responses to DCN vs. VCN Stimulation in a Mouse Model of the Auditory Brainstem Implant (ABI).

The auditory brainstem implant (ABI) is an auditory neuroprosthesis that provides hearing to deaf patients by electrically stimulating the cochlear nucleus (CN) of the brainstem. Whether such stimulation activates one or the other of the CN's two major subdivisions is not known. Here, we demonstrate clear response differences from the stimulation of the dorsal (D) vs. ventral (V) subdivisions of the CN in a mouse model of the ABI with a surface-stimulating electrode array. For the DCN, low levels of stimulation evoked multiunit responses in the inferior colliculus (IC) that were unimodally distributed with early latencies (avg. peak latency of 3.3 ms). However, high levels of stimulation evoked a bimodal distribution with the addition of a late latency response peak (avg. peak latency of 7.1 ms). For the VCN, in contrast, electrical stimulation elicited multiunit responses that were usually unimodal and had a latency similar to the DCN's late response. Local field potentials (LFP) from the IC showed components that correlated with early and late multiunit responses. Surgical cuts to sever the output of the DCN, the dorsal acoustic stria (DAS), gave insight into the origin of these early and late responses. Cuts eliminated early responses but had little-to-no effect on late responses. The early responses thus originate from cells that project through the DAS, such as DCN's pyramidal and giant cells. Late responses likely arise from the spread of stimulation from a DCN-placed electrode array to the VCN and could originate in bushy and/or stellate cells. In human ABI users, the spread of stimulation in the CN may result in abnormal response patterns that could hinder performance.

Preclinical upper limb neurorobotic platform to assess, rehabilitate, and develop therapies.

Numerous neurorehabilitative, neuroprosthetic, and repair interventions aim to address the consequences of upper limb impairments after neurological disorders. Although these therapies target widely different mechanisms, they share the common need for a preclinical platform that supports the development, assessment, and understanding of the therapy. Here, we introduce a neurorobotic platform for rats that meets these requirements. A four-degree-of-freedom end effector is interfaced with the rat's wrist, enabling unassisted to fully assisted execution of natural reaching and retrieval movements covering the entire body workspace. Multimodal recording capabilities permit precise quantification of upper limb movement recovery after spinal cord injury (SCI), which allowed us to uncover adaptations in corticospinal tract neuron dynamics underlying this recovery. Personalized movement assistance supported early neurorehabilitation that improved recovery after SCI. Last, the platform provided a well-controlled and practical environment to develop an implantable spinal cord neuroprosthesis that improved upper limb function after SCI.

Recent Advances in Encapsulation of Flexible Bioelectronic Implants: Materials, Technologies, and Characterization Methods.

Bioelectronic implantable systems (BIS) targeting biomedical and clinical research should combine long-term performance and biointegration in vivo. Here, recent advances in novel encapsulations to protect flexible versions of such systems from the surrounding biological environment are reviewed, focusing on material strategies and synthesis techniques. Considerable effort is put on thin-film encapsulation (TFE), and specifically organic-inorganic multilayer architectures as a flexible and conformal alternative to conventional rigid cans. TFE is in direct contact with the biological medium and thus must exhibit not only biocompatibility, inertness, and hermeticity but also mechanical robustness, conformability, and compatibility with the manufacturing of microfabricated devices. Quantitative characterization methods of the barrier and mechanical performance of the TFE are reviewed with a particular emphasis on water-vapor transmission rate through electrical, optical, or electrochemical principles. The integrability and functionalization of TFE into functional bioelectronic interfaces are also discussed. TFE represents a must-have component for the next-generation bioelectronic implants with diagnostic or therapeutic functions in human healthcare and precision medicine.

Prevention of the foreign body response to implantable medical devices by inflammasome inhibition.

SignificanceImplantable electronic medical devices (IEMDs) are used for some clinical applications, representing an exciting prospect for the transformative treatment of intractable conditions such Parkinson's disease, deafness, and paralysis. The use of IEMDs is limited at the moment because, over time, a foreign body reaction (FBR) develops at the device-neural interface such that ultimately the IEMD fails and needs to be removed. Here, we show that macrophage nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activity drives the FBR in a nerve injury model yet integration of an NLRP3 inhibitor into the device prevents FBR while allowing full healing of damaged neural tissue to occur.

NeuralTree: A 256-Channel 0.227-µJ/Class Versatile Neural Activity Classification and Closed-Loop Neuromodulation SoC.

Closed-loop neural interfaces with on-chip machine learning can detect and suppress disease symptoms in neurological disorders or restore lost functions in paralyzed patients. While high-density neural recording can provide rich neural activity information for accurate disease-state detection, existing systems have low channel counts and poor scalability, which could limit their therapeutic efficacy. This work presents a highly scalable and versatile closed-loop neural interface SoC that can overcome these limitations. A 256-channel time-division multiplexed (TDM) front-end with a two-step fast-settling mixed-signal DC servo loop (DSL) is proposed to record high-spatial-resolution neural activity and perform channel-selective brain-state inference. A tree-structured neural network (NeuralTree) classification processor extracts a rich set of neural biomarkers in a patient- and disease-specific manner. Trained with an energy-aware learning algorithm, the NeuralTree classifier detects the symptoms of underlying disorders (e.g., epilepsy and movement disorders) at an optimal energy-accuracy trade-off. A 16-channel high-voltage (HV) compliant neurostimulator closes the therapeutic loop by delivering charge-balanced biphasic current pulses to the brain. The proposed SoC was fabricated in 65nm CMOS and achieved a 0.227µJ/class energy efficiency in a compact area of 0.014mm2/channel. The SoC was extensively verified on human electroencephalography (EEG) and intracranial EEG (iEEG) epilepsy datasets, obtaining 95.6%/94% sensitivity and 96.8%/96.9% specificity, respectively. In-vivo neural recordings using soft µECoG arrays and multi-domain biomarker extraction were further performed on a rat model of epilepsy. In addition, for the first time in literature, on-chip classification of rest-state tremor in Parkinson's disease (PD) from human local field potentials (LFPs) was demonstrated.

Wireless closed-loop optogenetics across the entire dorsoventral spinal cord in mice.

Optoelectronic systems can exert precise control over targeted neurons and pathways throughout the brain in untethered animals, but similar technologies for the spinal cord are not well established. In the present study, we describe a system for ultrafast, wireless, closed-loop manipulation of targeted neurons and pathways across the entire dorsoventral spinal cord in untethered mice. We developed a soft stretchable carrier, integrating microscale light-emitting diodes (micro-LEDs), that conforms to the dura mater of the spinal cord. A coating of silicone-phosphor matrix over the micro-LEDs provides mechanical protection and light conversion for compatibility with a large library of opsins. A lightweight, head-mounted, wireless platform powers the micro-LEDs and performs low-latency, on-chip processing of sensed physiological signals to control photostimulation in a closed loop. We use the device to reveal the role of various neuronal subtypes, sensory pathways and supraspinal projections in the control of locomotion in healthy and spinal-cord injured mice.

Intrafascicular peripheral nerve stimulation produces fine functional hand movements in primates.

Restoring dexterous hand control is critical for people with paralysis. Approaches based on surface or intramuscular stimulation provide limited finger control, generate insufficient force to recover functional movements, and require numerous electrodes. Here, we show that intrafascicular peripheral electrodes could produce functional grasps and sustained forces in three monkeys. We designed an intrafascicular implantable electrode targeting the motor fibers of the median and radial nerves. Our interface selectively and reliably activated extrinsic and intrinsic hand muscles, generating multiple functional grips, hand opening, and sustained contraction forces for up to 2 months. We extended those results to a behaving monkey with transient hand paralysis and used intracortical signals to control simple stimulation protocols that enabled this animal to perform a functional grasping task. Our findings show that just two intrafascicular electrodes can generate a rich portfolio of dexterous and functional hand movements with important implications for clinical applicability.