3D Reconstruction of the Intracortical Volume Around a Hybrid Microelectrode Array

Extensive research using penetrating electrodes implanted in the central and peripheral nervous systems has been performed for many decades with significant advances made in recent years. While penetrating devices provide proximity to individual neurons in vivo, they suffer from declining performance over the course of months and often fail within a year. 2D histology studies using serial tissue sections have been extremely insightful in identifying and quantifying factors such as astroglial scar formation and neuronal death around the implant sites that may be contributing to failures. However, 2D histology has limitations in providing a holistic picture of the problems occurring at the electrode-tissue interface and struggles to analyze tissue below the electrode tips where the electrode tracks are no longer visible. In this study, we present 3D reconstruction of serial sections to overcome the limitations of 2D histological analysis. We used a cohort of software: XuvStitch, AutoAligner, and Imaris coupled with custom MATLAB programming to correct warping effects. Once the 3D image volume was reconstructed, we were able to use Imaris to quantify neuronal densities around the electrode tips of a hybrid microelectrode array incorporating Blackrock, Microprobes, and NeuroNexus electrodes in the same implant. This paper presents proof-of-concept and detailed methodological description of a technique which can be used to quantify neuronal densities in future studies of implanted electrodes

Doctor of Philosophy

dissertationBy enabling neuroprosthetic technologies, neural microelectrodes can greatly improve diagnostic and treatment options for millions of individuals living with limb loss, paralysis, and sensory and autonomic neural disorders. However, clinical use of these devices is restricted by the limited functional lifetimes of implanted electrodes, which are commonly less than a few years. One cause is the evolution of damage to dielectric encapsulation that insulates microelectrodes from the physiological environment. Fluid penetration and exposure to an aggressive immunological response over time may weaken encapsulating films and cause electrical shunting. This reduces electrode impedance, diverts electrical signal away from target tissue, and causes multi-channel crosstalk. To date, no neural microelectrode encapsulating material or design approach has reliably resolved this issue. We employ the parylene C-encapsulated Utah Electrode Array (UEA), a silicon-micromachined neural interface FDA-cleared for human use, to execute three aims that address this challenge through investigations of new materials, electrode designs, and testing methods. We first evaluate a novel bilayer encapsulating film comprised of atomic layer deposited Al2O3 and parylene C, testing this film using UEAs and devices with UEA-relevant topography. Contrasting with previous work employing simplified planar structures, the incorporation of neural electrode features on test structures revealed failure modes pointing to the dissolution of Al2O3 over time. Our results emphasize the need for dielectric coatings resistant to water degradation as well as test methods that take electrode features into account. In our second aim, we show through finite element modeling and aggressive in vitro testing that use of degenerately doped silicon as a conductive neural electrode material can mitigate the consequences of encapsulation damage, owing to the high electrochemical impedance of silicon. Our final aim compares oxidative in vitro aging to long-term in vivo material damages and finds clear evidence that such in vitro testbeds may help predict certain in vivo damage modes. By pairing this testing with absorption and emission spectroscopic characterization modalities, we identify contributors to material damage and future design solutions. Our results will inform future material and testing choices, to improve the resilience of neural electrode dielectric encapsulation and enhance the longevity of neuroprostheses

An investigation of extraocular and intraocular wireless communication techniques on a retinal prosthesis system

Retinitis Pigmentosa (RP) and Age-related Macular Degeneration (AMD) are two genetic ocular diseases that cause gradual visual impairments which will eventually lead to blindness as a result of damage in the retina. In the cases of people suffering from RP and AMD, it has been found out that 95% of the photoreceptors are damaged, while interestingly majority of the bipolar and ganglion cells that are responsible for the nerve stimulation remain intact. This is where a retinal prosthesis system comes into the picture. Retinal prosthesis is a prosthetic device that is aimed to assume the functionality of the damaged photoreceptors and produce stimulations to the bipolar and ganglion cells for a visual perception. Typically, a retinal prosthesis system comprises of two major components: an image capturing unit and an array of microelectrode. While a lot of studies have been conducted on each major component, the development of the wireless link between the two components has been mostly overlooked. It is clear that the two components are not physically connected and a data exchange is required between the two. This thesis aims to bridge the knowledge gap in this area by addressing the following research questions: “What is the most suitable frequency band for a wireless link in a retinal prosthesis system?” and “What kind of antenna would generate the most optimal performance under the constraints introduced by a retinal prosthesis system?

Implantable Electrodes for Upper Limb Prosthetic Control

This thesis describes a study investigating implantable interfaces with muscles and peripheral nerves. Current prostheses for upper limb amputees do not provide intuitive control over hand, wrist and elbow motion. By implanting electrodes for recording and stimulating onto muscles and into nerves in the amputation stump a greater number of control signals may be made available, signals which will be used to control dextrous hand movements. An implantable epimysial interface was developed using a bone-anchored device to hard-wire signals across the skin barrier. In a single ovine model pilot study the bone-anchor was implanted transtibially and the epimysial electrode was place superficially to m. peroneus teritus. Physiological signals were obtained over 12 weeks during treadmill walking. The external connector on the bone-anchor failed at 12 weeks, correlating with a drop in signal quality in an otherwise robust interface integrated with bone and skin tissue. The ovine bone-anchor model was repeated in 6 sheep for 19 weeks, with epimysial recordings made regularly. Increasing signal quality was seen during the study and was significantly greater from implanted electrodes compared with skin surface electrodes at 19 weeks (p = 0.016). Some complications with skin-implant integration were observed in proximally located implants. Crosstalk between muscles was assessed using pre-terminal nerve stimulation, and was found to be dependent upon muscle location and innervation. The ovine m. peroneus teritus model was used to assess recovery following targeted muscle reinnervation. Muscle signal recovery was observed approximately one month after surgery correlating with the start of functional recovery (assessed by force plate analysis). These studies indicate that a suitably modified bone-anchored device may be suitable for signal transmission in human patients, providing a stable, long-term solution to both prosthesis attachment and control. The potential of nerve interfaces for prosthetic control was investigated. The microchannel neural interface (MNI) was chosen because it overcomes limitations with other neural microarray designs: signal strength; cross-talk, and the locations of Nodes of Ranvier. MNIs confine regenerating nerves to small, ∼ 100 µm diameter, insulating tubes, this increases the length within which nerve signals can be recorded and amplifies the recorded signals. However, in vivo MNIs can become occluded by fibrosis that reduces or prevents axon regeneration. Two in vitro studies of neurocompatibility were carried out to investigate strategies for improving axon regeneration within microchannels. The first in vitro study compared the effect of different adsorbed endoneurial basement membrane proteins on PC-12 cell neurite extension on silicone substrates. The optimal protein coating concentrations for poly-D-lysine, collagen-IV and laminin-2,(-4) were determined. The optimal concentrations were compared with mixtures of basement membrane proteins, the effect of mixture coating order and constitution were investigated. It was found that endoneurial BM proteins significantly enhance neurite outgrowth compared with controls. Two coatings were suggested as most suited for improving neural regeneration within microchannels: a single layer coating of 10 µg/cm2 collagen-IV; and a mixed coating of 10 µg/cm2 collagen-IV, 1 µg/cm2 laminin-2,(-4), and 0.175 µg/cm2 nidogen-1. The second in vitro study investigated the effect of grooved, roughened and multi-scale silicone surfaces on on PC-12 cell neurite extension. Deeper, narrower grooves were shown to increase the extent of neurite alignment, while resulting in fewer, longer, neurites. Roughening surfaces was shown to increase the amount of protein (collagen-IV) which adsorbed from solution and increase the number of neurites each cell extended. Surfaces with multiscale topographies synergistically increased the number and length of neurites and guided neurite growth along the groove direction. MNIs were manufactured for in vivo testing. These MNIs were used to determine the effect of adsorbed endoneurial basement membrane proteins on nerve regeneration in vivo, but the multiscale topographies were not applied during manufacturing. Four alternative manufacturing methods were investigated and iterative improvements were made to create a stacked interface with multiple microchannel layers. Microchannel layers were created by laser patterning silicone and metal foil components, followed by plasma bonding to create a 3-dimensional structure with 150 µm deep, 200 µm wide microchannels. Electrode impedances of 27.2 ± 19.8 kΩ at 1kHz were achieved by DC etching. The method overcomes some current limitations on electrode connectivity and microchannel sealing, and may improve recording capabilities over single layer designs by increasing the ratio of electrodes to microchannels. Manufactured MNIs were tested in a rat sciatic nerve transection model. Following implantation nerves were allowed to regenerate for one and two months. First, suture and fibrin glue were compared as MNI fixation methods for one month, the nerve regenerated within the fibrin glue, outside the interface lumen, therefore sutures were chosen as a long term fixation method. The influence of endoneurial basement membrane protein coatings, identified previously, on nerve regeneration with MNIs was investigated. Nerves regenerated through the MNIs over two months and began to reinnervate the distal limb. Improvements in the sciatic function index were observed over two months, with no significant differences between protein coated and control interfaces. Some weak histological evidence for the use of protein coatings was found, with axon diameters increased distal to protein coated MNIs. Electromyographic and electroneurographic recordings demonstrated similar signal amplitudes to previous studies. In order to bring the research described in this thesis to clinical practice further engineering improvements to the design and manufacture of electrodes, which utilise materials or coatings to enhance neurocompatibility, is required. Avenues for further research are discussed and additional experiments and investigations are described. By combining developments in implantable muscle and nerve interfaces with surgical techniques and improvements in neurocompatibility the promise of upper limb prosthetic control may be realised