Interface-Mediated Neurogenic Signaling: The Impact of Surface Geometry and Chemistry on Neural Cell Behavior for Regenerative and Brain-Machine Interfacing Applications.

Nanomaterial advancements have driven progress in central and peripheral nervous system applications such as tissue regeneration and brain-machine interfacing. Ideally, neural interfaces with native tissue shall seamlessly integrate, a process that is often mediated by the interfacial material properties. Surface topography and material chemistry are significant extracellular stimuli that can influence neural cell behavior to facilitate tissue integration and augment therapeutic outcomes. This review characterizes topographical modifications, including micropillars, microchannels, surface roughness, and porosity, implemented on regenerative scaffolding and brain-machine interfaces. Their impact on neural cell response is summarized through neurogenic outcome and mechanistic analysis. The effects of surface chemistry on neural cell signaling with common interfacing compounds like carbon-based nanomaterials, conductive polymers, and biologically inspired matrices are also reviewed. Finally, the impact of these extracellular mediated neural cues on intracellular signaling cascades is discussed to provide perspective on the manipulation of neuron and neuroglia cell microenvironments to drive therapeutic outcomes.

A Library of Polymer-based Microelectrode Array Designs for Recording from the Brain of Different Animal Models.

Large-scale network recording technology is critical in linking neural activity to behavior. Stable, long-term recordings collected from behaving animals are the foundation for understanding neural dynamics and the plasticity of neural circuits. Penetrating microelectrode arrays (MEAs) can obtain high-resolution neural activity from different brain regions. However, ensuring the longevity of implantable devices and the consistency of neural signals over time remains one big challenge. A potential solution is to use flexible, polymer-based MEAs to minimize the foreign body response and prolong the lifetime of neural interfacing devices. Rodents and nonhuman primates (NHP) are commonly used animal models in neuroscience and neuroengineering studies. Specially designed MEAs that capture morphological features of different animal brains and various brain structures are powerful tools to simultaneously obtain neural activities from multiple brain regions. In this work, we develop a set of prototype designs of polymer MEAs that cover cortical, sub-cortical, and multiple brain regions of rodents and NHP.

Polymer Implantable Electrode Foundry: A shared resource for manufacturing polymer-based microelectrodes for neural interfaces.

Large scale monitoring of neural activity at the single unit level can be achieved via electrophysiological recording using implanted microelectrodes. While neuroscience researchers have widely employed chronically implanted electrode-based interfaces for this purpose, a commonly encountered limitation is loss of highly resolved signals arising from immunological response over time. Next generation electrode-based interfaces improve longitudinal signal quality using the strategy of stabilizing the device-tissue interface with microelectrode arrays constructed from soft and flexible polymer materials. The limited availability of such polymer microelectrode arrays has restricted access to a small number of researchers able to build their own custom devices or who have developed specific collaborations with engineering researchers who can produce them. Here, a new technology resource model is introduced that seeks to widely increase access to polymer microelectrode arrays by the neuroscience research community. The Polymer Implantable Electrode (PIE) Foundry provides custom and standardized polymer microelectrode arrays as well as training and guidance on best-practices for implantation and chronic experiments.

Characterization of thin film Parylene C device curvature and the formation of helices via thermoforming.

In microfabricated biomedical devices, flexible, polymer substrates are becoming increasingly preferred over rigid, silicon substrates because of their ability to conform to biological tissue. Such devices, however, are fabricated in a planar configuration, which results in planar devices that do not closely match the shape of most tissues. Thermoforming, a process which can reshape thermoplastic polymers, can be used to transform flat, thin film, polymer devices with patterned metal features into complex three-dimensional (3D) geometries. This process extends the use of planar microfabrication to achieve 3D shapes which can more closely interface with the body. Common shapes include spheres, which can conform to the shape of the retina; cones, which can be used as a sheath to interface with an insertion stylet; and helices, which can be wrapped around nerves, blood vessels, muscle fibers, or be used as strain relief feature. This work characterizes the curvature of thin film Parylene C devices with patterned metal features built with varying Parylene thicknesses and processing conditions. Device curvature is caused by film stress in each Parylene and metal layer, which is characterized experimentally and by a mathematical model which estimates the effects of device geometry and processing on curvature. Using this characterization, an optimized process to thermoform thin film Parylene C devices with patterned metal features into 0.25 mm diameter helices while preventing cracking in the polymer and metal was developed.

Making a case for endovascular approaches for neural recording and stimulation.

There are many electrode types for recording and stimulating neural tissue, most of which necessitate direct contact with the target tissue. These electrodes range from large, scalp electrodes which are used to non-invasively record averaged, low frequency electrical signals from large areas/volumes of the brain, to penetrating microelectrodes which are implanted directly into neural tissue and interface with one or a few neurons. With the exception of scalp electrodes (which provide very low-resolution recordings), each of these electrodes requires a highly invasive, open brain surgical procedure for implantation, which is accompanied by significant risk to the patient. To mitigate this risk, a minimally invasive endovascular approach can be used. Several types of endovascular electrodes have been developed to be delivered into the blood vessels in the brain via a standard catheterization procedure. In this review, the existing body of research on the development and application of endovascular electrodes is presented. The capabilities of each of these endovascular electrodes is compared to commonly used direct-contact electrodes to demonstrate the relative efficacy of the devices. Potential clinical applications of endovascular recording and stimulation and the advantages of endovascular versus direct-contact approaches are presented.

Acute in vivo Recording with a Generic Parylene Microelectrode Array Implanted with Dip-coating Method into the Rat Brain.

Flexible polymer-based microelectrode arrays (MEAs) can reduce tissue inflammation and foreign body response and greatly prolong the lifetime of neural implants. However, standard and customized polymer devices are only accessible to limited groups. To better promote the development and application of polymer MEAs, we have launched the Polymer Implantable Electrode (PIE) Foundry and developed a 64-channel Parylene C-based MEA with generic electrodes layout that can be used to record from both cortical and sub-cortical regions in rodents. In addition, a practical dip-coating protocol for the insertion of the flexible standard Parylene MEA is developed.

A portable multi-sensor module for monitoring external ventricular drains.

External ventricular drains (EVDs) are used clinically to relieve excess fluid pressure in the brain. However, EVD outflow rate is highly variable and typical clinical flow tracking methods are manual and low resolution. To address this problem, we present an integrated multi-sensor module (IMSM) containing flow, temperature, and electrode/substrate integrity sensors to monitor the flow dynamics of cerebrospinal fluid (CSF) drainage through an EVD. The impedimetric sensors were microfabricated out of biocompatible polymer thin films, enabling seamless integration with the fluid drainage path due to their low profile. A custom measurement circuit enabled automated and portable sensor operation and data collection in the clinic. System performance was verified using real human CSF in a benchtop EVD model. Impedimetric flow sensors tracked flow rate through ambient temperature variation and biomimetic pulsatile flow, reducing error compared with previous work by a factor of 6.6. Detection of sensor breakdown using novel substrate and electrode integrity sensors was verified through soak testing and immersion in bovine serum albumin (BSA). Finally, the IMSM and measurement circuit were tested for 53 days with an RMS error of 61.4 µL/min.

A comparison of insertion methods for surgical placement of penetrating neural interfaces.

Many implantable electrode arrays exist for the purpose of stimulating or recording electrical activity in brain, spinal, or peripheral nerve tissue, however most of these devices are constructed from materials that are mechanically rigid. A growing body of evidence suggests that the chronic presence of these rigid probes in the neural tissue causes a significant immune response and glial encapsulation of the probes, which in turn leads to gradual increase in distance between the electrodes and surrounding neurons. In recording electrodes, the consequence is the loss of signal quality and, therefore, the inability to collect electrophysiological recordings long term. In stimulation electrodes, higher current injection is required to achieve a comparable response which can lead to tissue and electrode damage. To minimize the impact of the immune response, flexible neural probes constructed with softer materials have been developed. These flexible probes, however, are often not strong enough to be inserted on their own into the tissue, and instead fail via mechanical buckling of the shank under the force of insertion. Several strategies have been developed to allow the insertion of flexible probes while minimizing tissue damage. It is critical to keep these strategies in mind during probe design in order to ensure successful surgical placement. In this review, existing insertion strategies will be presented and evaluated with respect to surgical difficulty, immune response, ability to reach the target tissue, and overall limitations of the technique. Overall, the majority of these insertion techniques have only been evaluated for the insertion of a single probe and do not quantify the accuracy of probe placement. More work needs to be performed to evaluate and optimize insertion methods for accurate placement of devices and for devices with multiple probes.

Bonding methods for chip integration with Parylene devices.

Flexible electronics require more compact interconnects for next-generation devices. Polymer devices can be bonded to integrated circuit chips, but combining flexible and rigid substrates poses unique technical challenges. Existing technologies either cannot achieve the density required for modern chips or employ specialized equipment and complex processes to do so. Here, we adapt several approaches to achieve fine-pitch bonding between rigid and flexible substrates including epoxy, ultrasonic wire, and anisotropic conductive film bonding and also introduce a novel technique called polymer ultrasonic on bump (PUB) bonding. Using Parylene C devices and various rigid substrates as our model testbed systems, we investigate these four methods across a range of bond pad size and pitch by measuring yield and resistance and by subjecting devices to thermomechanical reliability tests. We demonstrate that all methods are capable of bonding fine pitch interconnects (100 µm) at low temperature (<100 °C). Additionally, we focus on PUB bonding and join a packaged chip and a bare die to Parylene devices.

Acute in vivo testing of a polymer cuff electrode with integrated microfluidic channels for stimulation, recording, and drug delivery on rat sciatic nerve.

BACKGROUND: Extraneural cuffs are among the least invasive peripheral nerve interfaces as they remain outside the nerve. However, compared with more invasive interfaces, these electrodes may suffer from lower selectivity and sensitivity since the targeted nerve fibers are more distanced from the electrodes. NEW METHOD: A lyse-and-attract cuff electrode (LACE) was enabled by microfabrication and developed to improve selectivity and sensitivity while maintaining a cuff format. Its engineering design was described in previous work. LACE is a hybrid cuff that integrates both microelectrodes and microfluidic channels. The ultimate goal is to increase fascicular selectivity and sensitivity by focal delivery via the microchannels of (1) lysing agent to remove connective tissue separating electrodes from nerve fibers, and (2) neurotrophic factors to promote axonal sprouting of the exposed nerve fibers into microfluidic channels where electrodes are embedded. Here, we focus on demonstrating in vivo function of microfluidics and microelectrodes in an acute preparation in which we evaluate the ability to focally remove connective tissue and record and stimulate with microchannel-embedded microelectrodes neural activity in rat sciatic nerves. COMPARISON WITH EXISTING METHODS: While extraneural interfaces prioritize nerve health and intraneural interfaces prioritize functionality, LACE represents a new extraneural approach which could potentially excel at both aims. RESULTS: Surgical implantation demonstrate preservation of LACE function following careful and minimal handling. In vivo electrical evaluation demonstrates the ability of microelectrodes placed within microfluidic channels to successfully stimulate and record compound action potentials from rat sciatic nerve. Furthermore, collagen-rich epineurium was focally removed following infusion of collagenase via microchannels and confirmed via microscopy. CONCLUSION: The feasibility of using a cuff having integrated microelectrodes and microfluidics to stimulate, record, and deliver drug to focally lyse away the epineurium layer was demonstrated in acute experiments on rat sciatic nerve.

A Parylene Neural Probe Array for Multi-Region Deep Brain Recordings.

A Parylene C polymer neural probe array with 64 electrodes purposefully positioned across 8 individual shanks to anatomically match specific regions of the hippocampus was designed, fabricated, characterized, and implemented in vivo for enabling recording in deep brain regions in freely moving rats. Thin film polymer arrays were fabricated using surface micromachining techniques and mechanically braced to prevent buckling during surgical implantation. Importantly, the mechanical bracing technique developed in this work involves a novel biodegradable polymer brace that temporarily reduces shank length and consequently, increases its stiffness during implantation, therefore enabling access to deeper brain regions while preserving a low original cross-sectional area of the shanks. The resulting mechanical properties of braced shanks were evaluated at the benchtop. Arrays were then implemented in vivo in freely moving rats, achieving both acute and chronic recordings from the pyramidal cells in the cornu ammonis (CA) 1 and CA3 regions of the hippocampus which are responsible for memory encoding. This work demonstrated the potential for minimally invasive polymer-based neural probe arrays for multi-region recording in deep brain structures.

Recent advances in neural interfaces-Materials chemistry to clinical translation.

Implantable neural interfaces are important tools to accelerate neuroscience research and translate clinical neurotechnologies. The promise of a bidirectional communication link between the nervous system of humans and computers is compelling, yet important materials challenges must be first addressed to improve the reliability of implantable neural interfaces. This perspective highlights recent progress and challenges related to arguably two of the most common failure modes for implantable neural interfaces: (1) compromised barrier layers and packaging leading to failure of electronic components; (2) encapsulation and rejection of the implant due to injurious tissue-biomaterials interactions, which erode the quality and bandwidth of signals across the biology-technology interface. Innovative materials and device design concepts could address these failure modes to improve device performance and broaden the translational prospects of neural interfaces. A brief overview of contemporary neural interfaces is presented and followed by recent progress in chemistry, materials, and fabrication techniques to improve in vivo reliability, including novel barrier materials and harmonizing the various incongruences of the tissue-device interface. Challenges and opportunities related to the clinical translation of neural interfaces are also discussed.

A 512-Channel Multi-Layer Polymer-Based Neural Probe Array.

We present for the first time the design, fabrication, and preliminary bench-top characterization of a high-density, polymer-based penetrating microelectrode array, developed for chronic, large-scale recording in the cortices and hippocampi of behaving rats. We present two architectures for these targeted brain regions, both featuring 512 Pt recording electrodes patterned front-and-back on micromachined eight-shank arrays of thin-film Parylene C. These devices represent an order of magnitude improvement in both number and density of recording electrodes compared with prior work on polymer-based microelectrode arrays. We present enabling advances in polymer micro-machining related to lithographic resolution and a new method for back-side patterning of electrodes. In vitro electrochemical data verifies suitable electrode function and surface properties. Finally, we describe next steps toward the implementation of these arrays in chronic, large-scale recording studies in free-moving animal models.

A review for the peripheral nerve interface designer.

Informational density and relative accessibility of the peripheral nervous system make it an attractive site for therapeutic intervention. Electrode-based electrophysiological interfaces with peripheral nerves have been under development since the 1960s and, for several applications, have seen widespread clinical implementation. However, many applications require a combination of neural target resolution and stability which has thus far eluded existing peripheral nerve interfaces (PNIs). With the goal of aiding PNI designers in development of devices that meet the demands of next-generation applications, this review seeks to collect and present practical considerations and best practices which emerge from the literature, including both lessons learned during early PNI development and recent ideas. Fundamental and practical principles guiding PNI design are reviewed, followed by an updated and critical account of existing PNI designs and strategies. Finally, a brief survey of in vitro and in vivo PNI characterization methods is presented.

Continuous and bolus intraventricular topotecan prolong survival in a mouse model of leptomeningeal medulloblastoma.

Leptomeningeal metastasis remains a difficult clinical challenge. Some success has been achieved by direct administration of therapeutics into the cerebrospinal fluid (CSF) circumventing limitations imposed by the blood brain barrier. Here we investigated continuous infusion versus bolus injection of therapy into the CSF in a preclinical model of human Group 3 medulloblastoma, the molecular subgroup with the highest incidence of leptomeningeal disease. Initial tests of selected Group 3 human medulloblastoma cell lines in culture showed that D283 Med and D425 Med were resistant to cytosine arabinoside and methotrexate. D283 Med cells were also resistant to topotecan, whereas 1 µM topotecan killed over 99% of D425 Med cells. We therefore introduced D425 Med cells, modified to express firefly luciferase, into the CSF of immunodeficient mice. Mice were then treated with topotecan or saline in five groups: continuous intraventricular (IVT) topotecan via osmotic pump (5.28 µg/day), daily bolus IVT topotecan injections with a similar daily dose (6 µg/day), systemic intraperitoneal injections of a higher daily dose of topotecan (15 µg/day), daily IVT pumped saline and daily intraperitoneal injections of saline. Bioluminescence analyses revealed that both IVT topotecan treatments effectively slowed leptomeningeal tumor growth in the brains. Histological analysis showed that they were associated with localized brain necrosis, possibly due to backtracking of topotecan around the catheter. In the spines, bolus IVT topotecan showed a trend towards slower tumor growth compared to continuous (pump) IVT topotecan, as measured by bioluminescence. Both continuous and bolus topotecan IVT showed longer survival compared to other groups. Thus, both direct IVT topotecan CSF delivery methods produced better anti-medulloblastoma effect compared to systemic therapy at the dosages used here.

Application of Parylene-Based Flexible Multi-Electrode Array for Recording From Subcortical Brain Regions From Behaving Rats.

Obtaining multiple single-unit recordings in particular neural networks from behaving animals is crucial for the understanding of cognitive functions of the brain. Attaining stable, chronic recordings from the brain is also the foundation to develop effective cortical prosthetic devices. However, severe immune response caused by micromotion between stiff implants and surrounding brain tissue often limits the lifetime of penetrating, neural recording devices. To reduce the stiffness mismatch between recording devices and brain tissue, we developed a flexible, polymer based multi-electrode array for recording single neuron activities from the rat hippocampus, a major subcortical structure of the rat brain. Parylene C, a biocompatible polymer, was used as the structural and insulation material of the multi-electrode array. 64 platinum (Pt) recording electrodes were placed in groups along each shank to conform to the anatomical distribution of hippocampal principle neurons. The multi-electrode array was chronically implanted in three animals. After recovery, neural activity together with movement traces were collected from the behaving animals.

Techniques and Considerations in the Microfabrication of Parylene C Microelectromechanical Systems.

Parylene C is a promising material for constructing flexible, biocompatible and corrosion-resistant microelectromechanical systems (MEMS) devices. Historically, Parylene C has been employed as an encapsulation material for medical implants, such as stents and pacemakers, due to its strong barrier properties and biocompatibility. In the past few decades, the adaptation of planar microfabrication processes to thin film Parylene C has encouraged its use as an insulator, structural and substrate material for MEMS and other microelectronic devices. However, Parylene C presents unique challenges during microfabrication and during use with liquids, especially for flexible, thin film electronic devices. In particular, the flexibility and low thermal budget of Parylene C require modification of the fabrication techniques inherited from silicon MEMS, and poor adhesion at Parylene-Parylene and Parylene-metal interfaces causes device failure under prolonged use in wet environments. Here, we discuss in detail the promises and challenges inherent to Parylene C and present our experience in developing thin-film Parylene MEMS devices.

A review of implantable biosensors for closed-loop glucose control and other drug delivery applications.

Closed-loop drug delivery promises autonomous control of pharmacotherapy through the continuous monitoring of biomarker levels. For decades, researchers have strived for portable closed-loop systems capable of treating ambulatory patients with chronic conditions such as diabetes mellitus. After years of development, the first of these systems have left the laboratory and entered commercial use. This long-awaited advance reflects recent development of chronically stable implantable biosensors able to accurately measure biomarker levels in vivo. This review discusses the role of implantable biosensors in closed-loop drug delivery applications, with the intent to provide a resource for engineers and researchers studying such systems. We provide an overview of common biosensor designs and review the principle challenges in implementing long indwelling sensors: namely device sensitivity, selectivity, and lifetime. This review examines novel advances in transducer design, biological interface, and material biocompatibility, with a focus on recent academic and commercial work which provide successful strategies to overcome perennial challenges. This review focuses primarily on the topics of closed-loop glucose control and continuous glucose monitoring biosensors, which make up the overwhelming majority of published research in this area. We conclude with an overview of recent advances in closed-loop systems targeting applications outside blood glucose management.