Natalia Lago

A critical review of interfaces with the peripheral nervous system for the control of neuroprostheses and hybrid bionic systems

Abstract  Considerable scientific and technological efforts have been devoted to develop neuroprostheses and hybrid bionic systems that link the human nervous system with electronic or robotic prostheses, with the main aim of restoring motor and sensory functions in disabled patients. A number of neuroprostheses use interfaces with peripheral nerves or muscles for neuromuscular stimulation and signal recording. Herein, we provide a critical overview of the peripheral interfaces available and trace their use from research to clinical application in controlling artificial and robotic prostheses. The first section reviews the different types of non‐invasive and invasive electrodes, which include surface and muscular electrodes that can record EMG signals from and stimulate the underlying or implanted muscles. Extraneural electrodes, such as cuff and epineurial electrodes, provide simultaneous interface with many axons in the nerve, whereas intrafascicular, penetrating, and regenerative electrodes may contact small groups of axons within a nerve fascicle. Biological, technological, and material science issues are also reviewed relative to the problems of electrode design and tissue injury. The last section reviews different strategies for the use of information recorded from peripheral interfaces and the current state of control neuroprostheses and hybrid bionic systems.

Assessment of Biocompatibility of Chronically Implanted Polyimide and Platinum Intrafascicular Electrodes

Longitudinal intrafascicular electrodes (LIFEs) are electrodes designed to be placed inside the peripheral nerve to improve stimulation selectivity and to increase the recording signal-to-noise ratio. We evaluated the functional and morphological effects of either Pt wire LIFEs or polyimide-based thin-film LIFEs implanted in the rat sciatic nerve for 3 mo. The newly designed thin-film LIFEs are more flexible, can be micromachined and allow placement of more active electrode sites than conventional Pt LIFEs. Functional results at 1 mo indicated an initial decline in the nerve conduction velocity and in the amplitude of muscle responses, which recovered during the following 2 mo towards normal values. Morphological results showed that both types of LIFEs induced a mild scar response and a focal but chronic inflammatory reaction, which were limited to a small area around the electrode placed in the nerve. Both types of LIFEs can be considered biocompatible and cause reversible, minimal nerve damage

Metallothionein-1+2 protect the CNS after a focal brain injury

We have evaluated the physiological relevance of metallothionein-1+2 (MT-1+2) in the CNS following damage caused by a focal cryolesion onto the cortex. In comparison to normal mice, transgenic mice overexpressing the MT-1 isoform (TgMTI* mice) showed a significant decrease of the number of activated microglia/macrophage and of CD3+ T lymphocytes in the area surrounding the lesion, while astrocytosis was increased. The TgMTI* mice showed a diminished peripheral macrophage but not CD3 T cell response to the cryolesion. This altered inflammatory response produced a decreased expression of the proinflammatory cytokines IL-1beta, IL-6, and TNF-alpha and an increased expression of the growth factors bFGF, TGFbeta1, and VEGF in the TgMTI* mice relative to control mice, which might be related to the increased angiogenesis and regeneration of the parenchyma of the former mice. The overexpression of MT-1 dramatically reduced the cryolesion-induced oxidative stress and neuronal apoptosis. Remarkably, these effects were also obtained by the intraperitoneal administration of MT-2 to both normal and MT-1+2 knock-out mice. These results fully support the notion that MT-1+2 are essential in the CNS for coping with focal brain injury and suggest a potential therapeutic use of these proteins.

Astrocyte-targeted expression of IL-6 protects the CNSagainst a focal brain injury

The effect of CNS-targeted IL-6 gene expression has been thoroughly investigated in the otherwise nonperturbed brain but not following brain injury. Here we examined the impact of astrocyte-targeted IL-6 production in a traumatic brain injury (cryolesion) model using GFAP-IL6 transgenic mice. This study demonstrated that transgenic IL-6 production significantly increased wound healing following the cryolesion. Thus, at 20 days postlesion (dpl) the GFAP-IL6 mice showed almost complete wound healing compared to litter mate nontransgenic controls. It seems likely that a reduced inflammatory response in the long term could be responsible for this IL-6-related effect. Thus, while in the acute phase following cryolesion (1-6 dpl) the recruitment of macrophages and T lymphocytes was higher in GFAP-IL6 mice, at 10-20 dpl it was significantly reduced compared to controls. Reactive astrogliosis was also significantly increased up to but not including 20 dpl in the GFAP-IL6 mice. Oxidative stress as well as apoptotic cell death was significantly decreased throughout the time period studied in the GFAP-IL6 mice compared to controls. This could be linked to the altered inflammatory response as well as to the transgenic IL-6-induced increase of the antioxidant, neuroprotective proteins metallothionein-I + II. These results indicate that although in the brain the chronic astrocyte-targeted expression of IL-6 spontaneously induces an inflammatory response causing significant damage, during an acute neuropathological insult such as following traumatic injury, a clear neuroprotective role is evident.

Neurobiological Assessment of Regenerative Electrodes for Bidirectional Interfacing Injured Peripheral Nerves

Regenerative electrodes are designed to interface regenerated axons from a sectioned peripheral nerve. Applicability of regenerative electrodes depends on biocompatibility, success of axonal regeneration, secondary nerve damage, and adequacy of interface electronics. Polyimide sieve electrodes with 281 holes were chronically implanted in the severed sciatic nerve of 30 rats. Regeneration was successful in all the animals, with increasing numbers of regenerated myelinated fibers from 2 to 6 mo. However, constrictive axonopathy affected a few cases from 6 to 12 mo. postimplantation. A second electrode design with 571 holes and 27 ring electrodes was developed. The number of regenerated axons increased thanks to the larger open area. Recordings were obtained from a low proportion of electrodes on the sieve in response to distal stimulation. Difficulties for recording impulses with regenerative electrodes include the small size of regenerated axons, changes in membrane excitability and in target reconnection

Design, in vitro and in vivo assessment of a multi-channel sieve electrode with integrated multiplexer

This paper reports on the design, in vitro and in vivo investigation of a flexible, lightweight, polyimide based implantable sieve electrode with a hybrid assembly of multiplexers and polymer encapsulation. The integration of multiplexers enables us to connect a large number of electrodes on the sieve using few input connections. The implant assembly of the sieve electrode with the electronic circuitry was verified by impedance measurement. The 27 platinum electrodes of the sieve were coated with platinum black to reduce the electrode impedance. The impedance magnitude of the electrode sites on the sieve (geometric surface area 2,200 microm(2)) was |Z(f=1kHz)| = 5.7 kOmega. The sieve electrodes, encased in silicone, have been implanted in the transected sciatic nerve of rats. Initial experiments showed that axons regenerated through the holes of the sieve and reinnervated distal target organs. Nerve signals were recorded in preliminary tests after 3-7 months post-implantation.

Role of metallothioneins in peripheral nerve function and regeneration.

Abstract: The physiological role of the metallothionein (MT) family of proteins during peripheral nerve injury and regeneration was examined in Mt1+2 and Mt3 knockout (KO) mice. To this end, the right sciatic nerve was crushed, and the regeneration distance was evaluated by the pinch test 2-7 days postlesion (dpl) and electrophysiologically at 14 dpl. The quality of the regeneration was assessed by light microscopy and immunohistochemical methods. The results show that the regeneration distance was greater in the Mt3 KO than in the Mt1+2 KO mice, whereas control mice showed intermediate values. Moreover, the number of regenerating axons in the distal tibial nerve was significantly higher in Mt3KO mice than in the other two strains at 14 dpl. Immunoreactive profiles to protein gene product 9.5 were present in the epidermis and the sweat glands of the plantar skin of the hindpaw of the Mt3 KO group. The improved regeneration observed with the Mt3 KO mice was confirmed by compound nerve action potentials that were recorded from digital nerves at 14 dpl only in this group. We conclude that Mt3 normally inhibits peripheral nerve regeneration.

Effects of motor and sensory nerve transplants on amount and specificity of sciatic nerve regeneration

Nerve regeneration after complete transection does not allow for adequate functional recovery mainly because of lack of selectivity of target reinnervation. We assessed if transplanting a nerve segment from either motor or sensory origin may improve specifically the accuracy of sensory and motor reinnervation. For this purpose, the rat sciatic nerve was transected and repaired with a silicone guide containing a predegenerated segment of ventral root (VR) or dorsal root (DR), compared to a silicone guide filled with saline. Nerve regeneration and reinnervation was assessed during 3 months by electrophysiologic and functional tests, and by nerve morphology and immunohistochemistry against choline acetyltransferase (ChAT) for labeling motor axons. Functional tests showed that reinnervation was successful in all the rats. However, the two groups with a root allotransplant reached higher degrees of reinnervation in comparison with the control group. Group VR showed the highest reinnervation of muscle targets, whereas Group DR had higher levels of sensory reinnervation than VR and saline groups. The total number of regenerated myelinated fibers was similar in the three groups, but the number of ChAT+ fibers was slightly lower in the VR group in comparison with DR and saline groups. These results indicate that a predegenerated root nerve allotransplant enhances axonal regeneration, leading to faster and higher levels of functional recovery. Although there is not clear preferential reinnervation, regeneration of motor axons is promoted at early times by a motor graft, whereas reinnervation of sensory pathways is increased by a sensory graft.

Neurobiological evaluation of thin-film longitudinal intrafascicular electrodes as a peripheral nerve interface

Longitudinal intrafascicular electrodes (LIFEs) designed to be placed inside a peripheral nerve seem adequate devices for constituting a nerve-prosthesis interface. In this study we evaluated the functional and morphological effects of newly designed polyimide-based thin-film (tf-)LIFEs implanted in the rat sciatic nerve for 3 months. The tf-LIFEs used are highly flexible, and have 8 regularly spaced contacts, 4 at each side of the folded structure. Functional results at 1 month showed a mild decline in nerve conduction velocity and amplitude of muscle responses, which recovered during the following 2 months. Histological results showed a mild scar response around the intraneural electrode, but no signs of axonal loss and degeneration. Longitudinally implanted LIFEs behaved slightly better than transversally inserted LIFEs. Acute experiments proved that selective neural signal recording and nerve stimulation could be achieved from the different active sites in one tf-LIFE.

Evaluation of the long-term regenerative potential in an experimental nerve amputee model.

Abstract  In this study, we evaluated the long‐term maintenance of regenerated axons in an experimental nerve amputee model. The sciatic nerve of adult rats was transected and repaired with a silicone tube leaving a short gap; the distal nerve segment was again transected 10 mm distally and the distal stump either introduced in a capped silicone chamber (amputee group) or connected to denervated targets (tibial branch into the gastrocnemius muscle and peroneal nerve apposed to skin) (reinnervation group). Morphological studies were performed at 2.5, 6, and 9 months after surgery. In all cases, axons regenerated across the silicone tube and grew in the distal nerve segment. In the amputee group, the morphological results show the expected features of a neuroma that is formed when regenerating axons are prevented from reaching the end organs, with a large number of axonal profiles indicative of regenerative sprouting. The number of myelinated axons counted at the distal nerve was sustained over 9 months follow‐up, indicating that regenerated axons are maintained chronically. Immunohistochemical labeling showed maintained expression of choline acetyltransferase, calcitonin gene‐related peptide, and growth‐related peptides 43 in the distal neuroma at 6 and 9 months. Reconnection of the distal nerve to foreign targets mildly improved the pattern of nerve regeneration, decreasing the number of excessive sprouts. These results indicate that axons regenerated may be eventually interfaced with external input‐output systems over long time, even if ending in the absence of distal targets as will occur in amputee limbs.