Nerve Excitability and Neuropathic Pain is Reduced by BET Protein Inhibition After Spared Nerve Injury.

Neuropathic pain is a common disability produced by enhanced neuronal excitability after nervous system injury. The pathophysiological changes that underlie the generation and maintenance of neuropathic pain require modifications of transcriptional programs. In particular, there is an induction of pro-inflammatory neuromodulators levels, and changes in the expression of ion channels and other factors intervening in the determination of the membrane potential in neuronal cells. We have previously found that inhibition of the BET proteins epigenetic readers reduced neuroinflammation after spinal cord injury. Within the present study we aimed to determine if BET protein inhibition may also affect neuroinflammation after a peripheral nerve injury, and if this would beneficially alter neuronal excitability and neuropathic pain. For this purpose, C57BL/6 female mice underwent spared nerve injury (SNI), and were treated with the BET inhibitor JQ1, or vehicle. Electrophysiological and algesimetry tests were performed on these mice. We also determined the effects of JQ1 treatment after injury on neuroinflammation, and the expression of neuronal components important for the maintenance of axon membrane potential. We found that treatment with JQ1 affected neuronal excitability and mechanical hyperalgesia after SNI in mice. BET protein inhibition regulated cytokine expression and reduced microglial reactivity after injury. In addition, JQ1 treatment altered the expression of SCN3A, SCN9A, KCNA1, KCNQ2, KCNQ3, HCN1 and HCN2 ion channels, as well as the expression of the Na+/K+ ATPase pump subunits. In conclusion, both, alteration of inflammation, and neuronal transcription, could be the responsible epigenetic mechanisms for the reduction of excitability and hyperalgesia observed after BET inhibition. Inhibition of BET proteins is a promising therapy for reducing neuropathic pain after neural injury. PERSPECTIVE: Neuropathic pain is a common disability produced by enhanced neuronal excitability after nervous system injury. The underlying pathophysiological changes require modifications of transcriptional programs. This study notes that inhibition of BET proteins is a promising therapy for reducing neuropathic pain after neural injury.

Fascicular Topography of the Human Median Nerve for Neuroprosthetic Surgery.

One of the most sought-after applications of neuroengineering is the communication between the arm and an artificial prosthetic device for the replacement of an amputated hand or the treatment of peripheral nerve injuries. For that, an electrode is placed around or inside the median nerve to serve as interface for recording and stimulation of nerve signals coming from the fascicles that innervate the muscles responsible for hand movements. Due to the lack of a standard procedure, the electrode implantation by the surgeon is strongly based on intuition, which may result in poor performance of the neuroprosthesis because of the suboptimal location of the neural interface. To provide morphological data that can aid the neuroprosthetic surgeon with this procedure, we investigated the fascicular topography of the human median nerve along the forearm and upper arm. We first performed a description of the fascicular content and branching patterns along the length of the arm. Next we built a 3D reconstruction of the median nerve so we could analyze the fascicle morphological features in relation to the arm level. Finally, we characterized the motor content of the median nerve fascicles in the upper arm. Collectively, these results indicate that fascicular organization occurs in a short segment distal to the epicondyles and remains unaltered until the muscular branches leave the main trunk. Based on our results, overall recommendations based on electrode type and implant location can be drawn to help and aid the neuroprosthetic procedure. Invasive interfaces would be more convenient for the upper arm and the most proximal third of the forearm. Epineural electrodes seem to be most suitable for the forearm segment after fascicles have been divided from the main trunk.

Spatial and Functional Selectivity of Peripheral Nerve Signal Recording With the Transversal Intrafascicular Multichannel Electrode (TIME).

The selection of suitable peripheral nerve electrodes for biomedical applications implies a trade-off between invasiveness and selectivity. The optimal design should provide the highest selectivity for targeting a large number of nerve fascicles with the least invasiveness and potential damage to the nerve. The transverse intrafascicular multichannel electrode (TIME), transversally inserted in the peripheral nerve, has been shown to be useful for the selective activation of subsets of axons, both at inter- and intra-fascicular levels, in the small sciatic nerve of the rat. In this study we assessed the capabilities of TIME for the selective recording of neural activity, considering the topographical selectivity and the distinction of neural signals corresponding to different sensory types. Topographical recording selectivity was proved by the differential recording of CNAPs from different subsets of nerve fibers, such as those innervating toes 2 and 4 of the hindpaw of the rat. Neural signals elicited by sensory stimuli applied to the rat paw were successfully recorded. Signal processing allowed distinguishing three different types of sensory stimuli such as tactile, proprioceptive and nociceptive ones with high performance. These findings further support the suitability of TIMEs for neuroprosthetic applications, by exploiting the transversal topographical structure of the peripheral nerves.

Delaying discharge after the stimulus significantly decreases muscle activation thresholds with small impact on the selectivity: an in vivo study using TIME.

The number of devices for electrical stimulation of nerve fibres implanted worldwide for medical applications is constantly increasing. Stimulation charge is one of the most important parameters of stimulation. High stimulation charge may cause tissue and electrode damage and also compromise the battery life of the electrical stimulators. Therefore, the objective of minimizing stimulation charge is an important issue. Delaying the second phase of biphasic stimulation waveform may decrease the charge required for fibre activation, but its impact on stimulation selectivity is not known. This information is particularly relevant when transverse intrafascicular multichannel electrode (TIME) is used, since it has been designed to provide for high selectivity. In this in vivo study, the rat sciatic nerve was electrically stimulated using monopolar and bipolar configurations with TIME. The results demonstrated that the inclusion of a 100-µs delay between the cathodic and the anodic phase of the stimulus allows to reduce charge requirements by around 30 %, while only slightly affecting stimulation selectivity. This study shows that adding a delay to the typical stimulation waveform significantly ([Formula: see text]) reduces the charge required for nerve fibres activation. Therefore, waveforms with the delayed discharge phase are more suitable for electrical stimulation of nerve fibres.

Development of a neurotechnological system for relieving phantom limb pain using transverse intrafascicular electrodes (TIME).

Phantom limb pain (PLP) is a chronic condition that develops in the majority of amputees. The underlying mechanisms are not completely understood, and thus, no treatment is fully effective. Based on recent studies, we hypothesize that electrical stimulation of afferent nerves might alleviate PLP by giving sensory input to the patient if nerve fibers can be activated selectively. The critical component in this scheme is the implantable electrode structure. We present a review of a novel electrode concept to distribute highly selective electrode contacts over the complete cross section of a peripheral nerve to create a distributed activation of small nerve fiber ensembles at the fascicular level, the transverse intrafascicular multichannel nerve electrode (TIME). The acute and chronic implantations in a small animal model exhibited a good surface and structural biocompatibility as well as excellent selectivity. Implantation studies on large animal models that are closer to human nerve size and anatomical complexity have also been conducted. They proved implant stability and the ability to selectively activate nerve fascicles in a limited proximity to the implant. These encouraging results have opened the way forward for human clinical trials in amputees to investigate the effect of selective electrical stimulation on PLP.

Experimental validation of a hybrid computational model for selective stimulation using transverse intrafascicular multichannel electrodes.

Recently a hybrid model based on the finite element method and on a compartmental biophysical representation of peripheral nerve fibers and intraneural electrodes was developed founded on experimental physiological and histological data. The model appeared to be robust when dealing with uncertainties in parameter selection. However, an experimental validation of the findings provided by the model is required to fully characterize the potential of this approach. The recruitment properties of selective nerve stimulation using transverse intrafascicular multichannel electrodes (TIME) were investigated in this work in experiments with rats and were compared to model predictions. Animal experiments were performed using the same stimulation protocol as in the computer simulations in order to rigorously validate the model predictions and understand its limitations. Two different selectivity indexes were used, and new indexes for measuring electrode performance are proposed. The model predictions are in decent agreement with experimental results both in terms of recruitment curves and selectivity values. Results show that these models can be used for extensive studies targeting electrode shape design, active sites shape, and multipolar stimulation paradigms. From a neurophysiological point of view, the topographic organization of the rat sciatic nerve, on which the model was based, has been confirmed.

Comparative analysis of transverse intrafascicular multichannel, longitudinal intrafascicular and multipolar cuff electrodes for the selective stimulation of nerve fascicles.

The selection of a suitable nerve electrode for neuroprosthetic applications implies a trade-off between invasiveness and selectivity, wherein the ultimate goal is achieving the highest selectivity for a high number of nerve fascicles by the least invasiveness and potential damage to the nerve. The transverse intrafascicular multichannel electrode (TIME) is intended to be transversally inserted into the peripheral nerve and to be useful to selectively activate subsets of axons in different fascicles within the same nerve. We present a comparative study of TIME, LIFE and multipolar cuff electrodes for the selective stimulation of small nerves. The electrodes were implanted on the rat sciatic nerve, and the activation of gastrocnemius, plantar and tibialis anterior muscles was recorded by EMG signals. Thus, the study allowed us to ascertain the selectivity of stimulation at the interfascicular and also at the intrafascicular level. The results of this study indicate that (1) intrafascicular electrodes (LIFE and TIME) provide excitation circumscribed to the implanted fascicle, whereas extraneural electrodes (cuffs) predominantly excite nerve fascicles located superficially; (2) the minimum threshold for muscle activation with TIME and LIFE was significantly lower than with cuff electrodes; (3) TIME allowed us to selectively activate the three tested muscles when stimulating through different active sites of one device, both at inter- and intrafascicular levels, whereas selective activation using multipolar cuff (with a longitudinal tripolar stimulation configuration) was only possible for two muscles, at the interfascicular level, and LIFE did not activate selectively more than one muscle in the implanted nerve fascicle.

Biocompatibility of chronically implanted transverse intrafascicular multichannel electrode (TIME) in the rat sciatic nerve.

The transverse intrafascicular multichannel electrode (TIME) is intended to be transversally implanted in the peripheral nerve and to selectively interface subsets of axons in different fascicles within the same nerve. Two versions of TIME (TIME-2, TIME-3) were designed and tested for biocompatibility and safety in the sciatic nerve of the rat. TIME-2 was implanted in two groups: one group had only an acute implant and the second group had chronic implantation for 2 months; a third group was also chronically implanted with the TIME-3 version, designed to avoid the mechanical traction produced by muscles motion. We evaluated the functional and morphological effects of either TIME-2 or TIME-3 implanted in the rat sciatic nerve for 2 months. The results of the study indicate that implantation of the TIME-2 and TIME-3 devices in the rat sciatic nerve did not cause significant axonal loss or demyelination, as evidenced by the functional and histological results. The results of this study indicate that the TIME-2 and TIME-3 designs are biocompatible and safe after chronic implantation in a small peripheral nerve, such as the rat sciatic nerve.

A transverse intrafascicular multichannel electrode (TIME) to interface with the peripheral nerve.

Many different concepts of peripheral nerve interfaces have been developed over the past few decades and transferred into neuroscientific or clinical applications with varying degrees of success. In this study, we present a novel electrode design that transversally penetrates the peripheral nerve and is intended to selectively activate subsets of axons in different fascicles within the nerve. The "transverse intrafascicular multichannel electrode" (TIME) has been designed and fabricated using the micromachining and patterning of a polyimide substrate and insulation material and platinum electrode sites. In vitro characterization of the electrodes were carried out to determine the electrochemical transfer characteristics during recording and stimulation. Acute implantations were performed in the sciatic nerves of rats and recruitment curves were determined. Results indicated selective stimulation of different fascicles with graded recruitment. Future studies will address chronic implantation to investigate the reactions on the material-tissue interface and the long-term behavior and recruitment properties of the TIMEs.

Topographical distribution of motor fascicles in the sciatic-tibial nerve of the rat.

Knowledge of the intraneural topography of peripheral nerves may help to improve nerve repair after injuries and the selectivity of neural interfaces. We studied the fascicular pattern of motor fibers of the rat sciatic-tibial nerve. We carried out an anatomical dissection of the muscular tributaries of the tibial nerve in the leg. Immunohistochemistry against choline acetyltransferase was used to identify motor axons. Retrograde tracing allowed localization of the muscular fascicles at proximal levels of the sciatic trunk. The distribution of motor fibers in transverse section of the tibial nerve is not homogeneous; two clusters were identified, each one containing fibers of functionally related muscles. Retrograde tracing allowed for the identification of motor fascicles, each one well localized along the sciatic nerve. In the rat there is a somatotopic organization of the sciatic nerve, with muscular fascicles maintaining the same relative position along the entire nerve.