Flexible active electrode arrays with ASICs that fit inside the rat's spinal canal

Epidural spinal cord electrical stimulation (ESCS) has been used as a means to facilitate locomotor recovery in spinal cord injured humans. Electrode arrays, instead of conventional pairs of electrodes, are necessary to investigate the effect of ESCS at different sites. These usually require a large number of implanted wires, which could lead to infections. This paper presents the design, fabrication and evaluation of a novel flexible active array for ESCS in rats. Three small (1.7 mm2) and thin (100 μm) application specific integrated circuits (ASICs) are embedded in the polydimethylsiloxane-based implant. This arrangement limits the number of communication tracks to three, while ensuring maximum testing versatility by providing independent access to all 12 electrodes in any configuration. Laser-patterned platinum-iridium foil forms the implant’s conductive tracks and electrodes. Double rivet bonds were employed for the dice microassembly. The active electrode array can deliver current pulses (up to 1 mA, 100 pulses per second) and supports interleaved stimulation with independent control of the stimulus parameters for each pulse. The stimulation timing and pulse duration are very versatile. The array was electrically characterized through impedance spectroscopy and voltage transient recordings. A prototype was tested for long term mechanical reliability when subjected to continuous bending. The results revealed no track or bond failure. To the best of the authors’ knowledge, this is the first time that flexible active electrode arrays with embedded electronics suitable for implantation inside the rat’s spinal canal have been proposed, developed and tested in vitro

An Implantable Versatile Electrode-Driving ASIC for Chronic Epidural Stimulation in Rats

This paper presents the design and testing of an electrode driving application specific integrated circuit (ASIC) intended for epidural spinal cord electrical stimulation in rats. The ASIC can deliver up to 1 mA fully programmable monophasic or biphasic stimulus current pulses, to 13 electrodes selected in any possible configuration. It also supports interleaved stimulation. Communication is achieved via only 3 wires. The current source and the control of the stimulation timing were kept off-chip to reduce the heat dissipation close to the spinal cord. The ASIC was designed in a 0.18- \mu m high voltage CMOS process. Its output voltage compliance can be up to 25 V. It features a small core area ( {< } 0.36 mm ^{2} ) and consumes a maximum of 114 \mu W during a full stimulation cycle. The layout of the ASIC was developed to be suitable for integration on the epidural electrode array, and two different versions were fabricated and electrically tested. Results from both versions were almost indistinguishable. The performance of the system was verified for different loads and stimulation parameters. Its suitability to drive a passive epidural 12-electrode array in saline has also been demonstrated

Advances in Scalable Implantable Systems for Neurostimulation Using Networked ASICs

Neurostimulation is a known method for restoring lost functions to neurologically impaired patients. This paper describes recent advances in scalable implantable stimulation systems using networked application specific integrated circuits (ASICs). It discusses how they can meet the ever-growing demand for high-density neural interfacing and long-term reliability. A detailed design example of an implantable (inductively linked) scalable stimulation system for restoring lower limb functions in paraplegics after spinal cord injury is presented. It comprises a central hub implanted at the costal margin and multiple Active Books which provide the interface for stimulating nerve roots in the cauda equina. A 16-channel stimulation system using four Active Books is demonstrated. Each Active Book has an embedded ASIC, which is responsible for initiating stimulus current to the electrodes. It also ensures device safety by monitoring temperature, humidity, and peak electrode voltage during stimulation. The implant hub was implemented using a microcontroller-based circuit. The ASIC in the Active Book was fabricated using XFAB’s 0.6-µm high-voltage CMOS process. The stimulation system does not require an accurate reference clock in the implant. Measured results are provided

Afferent information modulates spinal network activity in vitro and in preclinical animal models

Primary afferents are responsible for the transmission of peripheral sensory information to the spinal cord. Spinal circuits involved in sensory processing and in motor activity are directly modulated by incoming input conveyed by afferent fibres. Current neurorehabilitation exploits primary afferent information to induce plastic changes within lesioned spinal circuitries. Plasticity and neuromodulation promoted by activity-based interventions are suggested to support both the functional recovery of locomotion and pain relief in subjects with sensorimotor disorders. The present study was aimed at assessing spinal modifications mediated by afferent information. At the beginning of my PhD project, I adopted a simplified in vitro model of isolated spinal cord from the newborn rat. In this preparation, dorsal root (DR) fibres were repetitively activated by delivering trains of electrical stimuli. Responses of dorsal sensory-related and ventral motor-related circuits were assessed by extracellular recordings. I demonstrated that electrostimulation protocols able to activate the spinal CPG for locomotion, induced primary afferent hyperexcitability, as well. Thus, evidence of incoming signals in modulating spinal circuits was provided. Furthermore, a robust sensorimotor interplay was reported to take place within the spinal cord. I further investigated hyperexcitability conditions in a new in vivo model of peripheral neuropathic pain. Adult rats underwent a surgical procedure where the common peroneal nerve was crushed using a calibrated nerve clamp (modified spared nerve injury, mSNI). Thus, primary afferents of the common peroneal nerve were activated through the application of a noxious compression, which presumably elicited ectopic activity constitutively generated in the periphery. One week after surgery, animals were classified into two groups, with (mSNI+) and without (mSNI-) tactile hypersensitivity, based on behavioral tests assessing paw withdrawal threshold. Interestingly, the efficiency of the mSNI in inducing tactile hypersensitivity was halved with respect to the classical SNI model. Moreover, mSNI animals with tactile hypersensitivity (mSNI+) showed an extensive neuroinflammation within the dorsal horn, with activated microglia and astrocytes being significantly increased with respect to mSNI animals without tactile hypersensitivity (mSNI-) and to sham-operated animals. Lastly, RGS4 (regulator of G protein signaling 4) was reported to be enhanced in lumbar dorsal root ganglia (DRGs) and dorsal horn ipsilaterally to the lesion in mSNI+ animals. Thus, a new molecular marker was demonstrated to be involved in tactile hypersensitivity in our preclinical model of mSNI. Lastly, we developed a novel in vitro model of newborn rat, where hindlimbs were functionally connected to a partially dissected spinal cord and passively-driven by a robotic device (Bipedal Induced Kinetic Exercise, BIKE). I aimed at studying whether spinal activity was influenced by afferent signals evoked during passive cycling. I first demonstrated that BIKE could actually evoke an afferent feedback from the periphery. Then, I determined that spinal circuitries were differentially affected by training sessions of different duration. On one side, a short exercise session could not directly activate the locomotor CPG, but was able to transiently facilitate an electrically-induced locomotor-like activity. Moreover, no changes in reflex or spontaneous activity of dorsal and ventral networks were promoted by a short training. On the other side, a long BIKE session caused a loss in facilitation of spinal locomotor networks and a depression in the area of motor reflexes. Furthermore, activity in dorsal circuits was long-term enhanced, with a significant increase in both electrically-evoked and spontaneous antidromic discharges. Thus, the persistence of training-mediated effects was different, with spinal locomotor circuits being only transiently modulated, whereas dorsal activity being strongly and stably enhanced. Motoneurons were also affected by a prolonged training, showing a reduction in membrane resistance and an increase in the frequency of post-synaptic currents (PSCs), with both fast- and slow-decaying synaptic inputs being augmented. Changes in synaptic transmission onto the motoneuron were suggested to be responsible for network effects mediated by passive training. In conclusion, I demonstrated that afferent information might induce changes within the spinal cord, involving both neuronal and glial cells. In particular, spinal networks are affected by incoming peripheral signals, which mediate synaptic, cellular and molecular modifications. Moreover, a strong interplay between dorsal and ventral spinal circuits was also reported. A full comprehension of basic mechanisms underlying sensory-mediated spinal plasticity and bidirectional interactions between functionally different spinal networks might lead to the development of neurorehabilitation strategies which simultaneously promote locomotor recovery and pain relief

An Electrophysiological Study Of Voluntary Movement and Spinal Cord Injury

Voluntary movement is generated from the interaction between neurons in our brain and the neurons in our spinal cord that engage our muscles. A spinal cord injury destroys the connection between these two regions, but parts of their underlying neural circuits survive. A new class of treatment (the brain-machine interface) takes advantage of this fact by either a) recording neural activity from the brain and predicting the intended movement (neural prosthetics) or b) stimulating neural activity in the spinal cord to facilitate muscle activity (spinal stimulation). This thesis covers new research studying the brain-machine interface and its application for spinal injury. First, the electrical properties of the microelectrode (the main tool of the brain-machine interface) are studied during deep brain recording and stimulation. This work shows that the insulation coating the electrode forms a capacitor with the surrounding neural tissue. This capacitance causes large spikes of voltage in the surrounding tissue during deep brain stimulation, which will cause electrical artifacts in neural recordings and may damage the surrounding neurons. This work also shows that a coaxially shielded electrode will block this effect. Second, the activity of neurons in the parietal cortex is studied during hand movements, which has applications for neural prosthetics. Prior work suggests that the parietal cortex encodes a state-estimator [1], which combines sensory feedback with the internal efference copy to predict the state of the hand. To test this idea, we used a visual lag to misalign sensory feedback from the efference copy. The expectation was that a state-estimator would unknowingly combine the delayed visual feedback with the current efference information, resulting in incorrect predictions of the hand. Our results show a drop in correlation between neural activity in the parietal cortex and hand movement during a visual lag, supporting the idea that the parietal cortex encodes a state-estimator. This correlation gradually recovers over time, showing that parietal cortex is adaptive to sensory delays. Third, while the intention of spinal stimulation was to interact locally with neural circuits in the spinal cord, results from the clinic show that electrical stimulation of the lumbosacral enlargement enables paraplegic patients to regain voluntary movement of their legs [2]. This means that spinal stimulation facilitates communication across an injury site. To further study this effect, we developed a new behavioral task in the rodent. Rats were trained to kick their right hindlimb in response to an auditory cue. The animals then received a spinal injury that caused paraplegia. After injury, the animals recovered the behavior (they could kick in response to the cue), but only during spinal stimulation. Their recovered behavior was slower and more stereotyped than their pre-injury response. Administering quipazine to these rodents disrupted their ability to respond to the cue, suggesting that serotonin plays an important role in the recovered pathway. This work proves that the new behavioral task is a successful tool for studying the recovery of voluntary movement. Future work will combine cortical recordings with this behavioral task in the rodent to study plasticity in the nervous system and improve treatment of spinal cord injuries. [1] Mulliken, Grant H., Sam Musallam, and Richard A. Andersen. "Forward estimation of movement state in posterior parietal cortex." Proceedings of the National Academy of Sciences105.24 (2008): 8170-8177. [2] Harkema, Susan, et al. "Effect of epidural stimulation of the lumbosacral spinal cord on voluntary movement, standing, and assisted stepping after motor complete paraplegia: a case study." The Lancet 377.9781 (2011): 1938-1947.</p

Closed-loop approaches for innovative neuroprostheses

The goal of this thesis is to study new ways to interact with the nervous system in case of damage or pathology. In particular, I focused my effort towards the development of innovative, closed-loop stimulation protocols in various scenarios: in vitro, ex vivo, in vivo

Neurostimulateur hautement intégré et nouvelle stratégie de stimulation pour améliorer la miction chez les paraplégiques

RÉSUMÉ Une lésion de la moelle épinière est un problème dévastateur médicalement et socialement. Pour la population des États-Unis seulement, il y a près de 10 000 nouveaux cas chaque année. A cause des nombreux types de lésions possibles, divers degrés de dysfonctionnement du bas appareil urinaire peuvent en découler. Une lésion est dite complète lors d’une perte totale des fonctions sensorielles et motrices volontaires en dessous du niveau de la lésion. Une lésion incomplète implique que certaines activités sensorielles et/ou motrices soient encore présentes. Si la lésion se produit au dessus du cône médullaire, la vessie développera une hyperréflexie qui se manifeste par des contractions réflexes non-inhibées. Ces contractions peuvent être accompagnées d’une augmentation de l’activité du sphincter externe. Par conséquent, cela mène à un état d’obstruction fonctionnelle de la vessie, qui induit une forte pression intravésicale à chacune des contractions réflexes et qui peut potentiellement endommager le haut appareil urinaire. Dans ce contexte, la neurostimulation est l'une des techniques les plus prometteuses pour la réhabilitation de la vessie chez les patients ayant subi une lésion de la moelle épinière. Le seul neurostimulateur implantable commercialisé, ciblant l'amélioration de la miction et ayant obtenu des résultats satisfaisants, nécessite une rhizotomie (section de certains nerfs) afin de réduire la dyssynergie entre la vessie et le sphincter. Cependant, la rhizotomie est irréversible et peut abolir les réflexes sexuels, de défécation ainsi que les sensations sacrales si encore présents dans le cas de lésions incomplètes. Afin d'éviter la rhizotomie, nous proposons une nouvelle stratégie de stimulation multi-site appliquée aux racines sacrées, et basée sur le blocage de la conduction des nerfs à l'aide d'une stimulation à haute fréquence comme alternative à la rhizotomie. Cette approche permettrait une meilleure miction en augmentant sélectivement la contraction de la vessie et en diminuant la dyssynergie. Huit expériences en phase aigüe ont étés menées sur des chiens pour vérifier la réponse de la vessie et du sphincter urétral externe à la stratégie de stimulation proposée. Le blocage à haute-fréquence (1 kHz) combiné à la stimulation basse-fréquence (30 Hz), a augmenté la différence de pression intra-vésicale/intra-urétrale moyenne jusqu'à 53 cmH2O et a réduit la pression intra-urétrale moyenne jusqu'à hauteur de 86 % relativement au niveau de référence. Dans l’objectif de tester la stratégie de neurostimulation proposée avec des expériences animales en phase chronique, un dispositif de neurostimulation implantable est requis. Un prototype discret implémentant cette stratégie de stimulation a été réalisé en utilisant uniquement des composants discrets disponibles commercialement. Ce prototype est capable de générer des impulsions à une fréquence aussi basse que 18 Hz tout en générant simultanément une forme d’onde alternative à une fréquence aussi haute que 8.6 kHz, et ce sur de multiples canaux. Lorsque tous les étages de stimulation et leurs différentes sorties sont activés avec des fréquences d’impulsions (2 mA, 217 μs) et de sinusoïdes de 30 Hz et 1 kHz respectivement, la consommation de puissance totale est autour de 4.5 mA (rms). Avec 50 mW de puissance inductive disponible par exemple et 4.5 mA de consommation de courant, le régulateur haute-tension peut être réglé à 10 V permettant ainsi une stimulation de 2 mA avec une impédance nerf-électrode de 4.4 kΩ. Le nombre effectif de sorties activées et le maximum réalisable des paramètres de stimulation sont limités par l’énergie disponible fournie par le lien inductif et l’impédance des interfaces nerf-électrode. Cependant, une plus grande intégration du neurostimulateur devient de plus en plus nécessaire à des fins de miniaturisation, de réduction de consommation de puissance, et d’augmentation du nombre de canaux de stimulation. Comme première étape vers une intégration totale, nous présentons la conception d’un neurostimulateur hautement intégré et qui peut être assemblé sur un circuit imprimé de 21 mm de diamètre. Le prototype est basé sur trois circuits intégrés, dédiés et fabriqués en technologie CMOS haute-tension, ainsi qu’un FPGA miniature à faible puissance et disponible commercialement. En utilisant une approche basée sur un abaisseur de tension, où la tension induite est laissée libre jusqu’à 20 V, l’étage d’entrée de récupération de puissance inductive et de données est totalement intégré.----------ABSTRACT Spinal cord injury (SCI) is a devastating condition medically and socially. For the population of USA only, the incidence is around 10 000 new cases per year. SCI leads to different degrees of dysfunction of the lower urinary tract due to a large variety of possible lesions. With a complete lesion, there is a complete loss of sensory and motor control below the level of lesion. An incomplete lesion implies that some sensory and/or motor activity is still present. Most patients with suprasacral SCI suffer from detrusor over-activity (DO) and detrusor sphincter dyssynergia (DSD). DSD leads to high intravesical pressure, high residual urine, urinary tract infection, and deterioration of the upper urinary tract. In this context, neurostimulation is one of the most promising techniques for bladder rehabilitation in SCI patients. The only commercialized implantable neurostimulator aiming for improved micturition and having obtained satisfactory results requires rhizotomy to reduce DSD. However, rhizotomy is irreversible and may abolish sexual and defecation reflexes as well as sacral sensations, if still present in case of incomplete SCI. In order to avoid rhizotomy, we propose a new multisite stimulation strategy applied to sacral roots, and based on nerve conduction blockade using high-frequency stimulation as an alternative to rhizotomy. This approach would allow a better micturition by increasing bladder contraction selectively and decreasing dyssynergia. Eight acute dog experiments were carried out to verify the bladder and the external urethral sphincter responses to the proposed stimulation strategy. High-frequency blockade (1 kHz) combined with low-frequency stimulation (30 Hz) increased the average intravesical-intraurethral pressure difference up to 53 cmH2O and reduced the average intraurethral pressure with respect to baseline by up to 86 %. To test the proposed neurostimulation strategy during chronic animal experiments, an implantable neurostimulateur is required. A discrete prototype implementing the proposed stimulation strategy has been designed using commercially available discrete components. This prototype is capable of generating a low frequency pulse waveform as low as 18 Hz with a simultaneous high frequency alternating waveform as high as 8.6 kHz, and that over different and multiple channels

A Study of Techniques and Mechanisms of Vagus Nerve Stimulation for Treatment of Inflammation

Vagus nerve stimulation (VNS) has been on the forefront of inflammatory disorder research for the better part of the last three decades and has yielded many promising results. There remains, however, much debate about the actual biological mechanisms of such treatments, as well as, questions about inconsistencies in methods used in many research efforts. In this work, I identify shortcomings in past VNS methods and submit new developments and findings that can progress the research community towards more selective and relevant VNS research and treatments. In Aim 1, I present the most recent advancements in the capabilities of our fully implantable Bionode stimulation device platform for use in VNS studies to include stimulation circuitry, device packaging, and stimulation cuff design. In Aim 2, I characterize the inflammatory cytokine response of rats to intraperitoneally injected endotoxin utilizing new data analysis methods and demonstrate the modulatory effects of VNS applied by the Bionode stimulator to subdiaphragmatic branches of the left vagus nerve in an acute study. In Aim 3, using fully implanted Bionode devices, I expose a previously unidentified effect of chronically cuffing the left cervical vagus nerve to suppress efferent Fluorogold transport and cause unintended attenuation to physiological effects of VNS. Finally, in accordance with our findings from Aims 1, 2, and 3, I present results from new and promising techniques we have explored for future use of VNS in inflammation studies

The mechanisms of GABAergic signalling in the peripheral pain pathway

Peripheral pain pathway plays a crucial role in how pain is perceived and felt. The dorsal root ganglia (DRG) which house the primary sensory neurons have become the focus of many emerging pain studies due to its potential as a functional structure in controlling pain transmission, and not only for producing proteins and providing nutrients essential for neuron survival. The major inhibitory neurotransmitter in the nervous system, GABA has been shown to play a significant role in this regard. Within the present study, the mechanism of GABA release within DRG neurons was investigated by studying the expression of vesicular GABA transporter (VGAT) in the DRG neurons. VGAT was highly expressed in the DRG neuron somata. The VGAT-positive neurons also expressed markers of subpopulations of DRG neurons, including those involved in nociception. The availability of VGAT luminal (VGAT-C) and cytoplasmic (VGAT-N) domains were utilised to investigate the mechanism of GABA release in a live DRG neuron culture. This mechanism involves the recycling process of vesicles following their exocytosis. Imaging of the internalization of VGAT-C domain during vesicle recycling indicates GABA is released via exocytosis and has both, tonic and activity-dependent components. Using the in vivo electrophysiological recordings, neuronal firing in the spinal nerve and dorsal branches of the peripheral nerve (before and after the DRG, respectively), was investigated. These data revealed existence of a ‘filter’ in the DRG that decreased the frequency of the neuronal firing passing through the DRG. This filtering effect was overcome by bicuculline, a GABAA receptor antagonist indicating the role of GABAA receptor in peripheral pain pathway. This role of GABAA receptor was also supported by the decrease in GABAA receptor activation in the presence of bicuculline in DRG neurons co-cultured with HEK293 cells. In sum, in the DRG, GABA is liberated into the interneuronal space via Ca2+-dependent vesicular exocytosis, which in turn acts on GABAA receptors. This GABAergic signalling is responsible for filtering the action potentials from the periphery to the central terminals in the spinal cord. These findings identify and further characterize peripheral ‘gate’ within the somatosensory system