preparation-of-rat-sciatic-nerve-for-ex-vivo-neurophysiology

Preparation of Rat Sciatic Nerve for <em>Ex Vivo</em> Neurophysiology

Ex vivo preparations enable the study of many neurophysiological processes in isolation from the rest of the body while preserving local tissue structure. ...

Imperial College London

Biological components of perilymph affect the electrical performance of cochlear implants. Understanding the perilymph composition of common animal models will improve the understanding of this impact and improve the interpretation of results from animal studies and how it relates to humans.

Comparing perilymph proteomes across species.

Nerve block waveforms require the passage of large amounts of electrical energy at the neural interface for extended periods of time. It is desirable that such waveforms be applied chronically, consistent with the treatment of protracted immune conditions, however current metal electrode technologies are limited in their capacity to safely deliver ongoing stable blocking waveforms. Conductive hydrogel (CH) electrode coatings have been shown to improve the performance of conventional bionic devices, which use considerably lower amounts of energy than conventional metal electrodes to replace or augment sensory neuron function. In this study the application of CH materials was explored, using both a commercially available platinum iridium (PtIr) cuff electrode array and a novel low-cost stainless steel (SS) electrode array. The CH was able to significantly increase the electrochemical performance of both array types. The SS electrode coated with the CH was shown to be stable under continuous delivery of 2 mA square pulse waveforms at 40,000 Hz for 42 days. CH coatings have been shown as a beneficial electrode material compatible with long-term delivery of high current, high energy waveforms.

Conductive Hydrogel Electrodes for Delivery of Long-Term High Frequency Pulses.

Promoting nerve regeneration requires engineering cellular carriers to physically and biochemically support neuronal growth into a long lasting functional tissue. This study systematically evaluated the capacity of a biosynthetic poly(vinyl alcohol) (PVA) hydrogel to support growth and differentiation of co-encapsulated neurons and glia. A significant challenge is to understand the role of the dynamic degradable hydrogel mechanical properties on expression of relevant cellular morphologies and function. It was hypothesised that a carrier with mechanical properties akin to neural tissue will provide glia with conditions to thrive, and that glia in turn will support neuronal survival and development. PVA co-polymerised with biological macromolecules sericin and gelatin (PVA-SG) and with tailored nerve tissue-like mechanical properties were used to encapsulate Schwann cells (SCs) alone and subsequently a co-culture of SCs and neural-like PC12s. SCs were encapsulated within two PVA-SG gel variants with initial compressive moduli of 16 kPa and 2 kPa, spanning a range of reported mechanical properties for neural tissues. Both hydrogels were shown to support cell viability and expression of extracellular matrix proteins, however, SCs grown within the PVA-SG with a higher initial modulus were observed to present with greater physiologically relevant morphologies and increased expression of extracellular matrix proteins. The higher modulus PVA-SG was subsequently shown to support development of neuronal networks when SCs were co-encapsulated with PC12s. The lower modulus hydrogel was unable to support effective development of neural networks. This study demonstrates the critical link between hydrogel properties and glial cell phenotype on development of functional neural tissues. STATEMENT OF SIGNIFICANCE: Hydrogels as platforms for tissue regeneration must provide encapsulated cellular progenitors with physical and biochemical cues for initial survival and to support ongoing tissue formation as the artificial network degrades. While most research focuses on tailoring scaffold properties to suit neurons, this work aims to support glia SCs as the key cellular component that physically and biochemically supports the neuronal network. The challenge is to modify hydrogel properties to support growth and development of multiple cell types into a neuronal network. Given SCs ability to respond to substrate mechanical properties, the significance of this work lies in understanding the relationship between dynamic hydrogel mechanical properties and glia SCs development as the element that enables formation of mature, differentiated neural networks.

Tissue engineered hydrogels supporting 3D neural networks.

Flexible polymeric bioelectronics have the potential to address the limitations of metallic electrode arrays by minimizing the mechanical mismatch at the device-tissue interface for neuroprosthetic applications. This work demonstrates the straightforward fabrication of fully organic electrode arrays based on conductive elastomers (CEs) as a soft, flexible and stretchable electroactive composite material. CEs were designed as hybrids of polyurethane elastomers (PU) and poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS), with the aim of combining the electrical properties of PEDOT:PSS with the mechanical compliance of elastomers. CE composites were fabricated by solvent casting of PEDOT:PSS dispersed in dissolved PU at different conductive polymer (CP) loadings, from 5 wt% to 25 wt%. The formation of PEDOT:PSS networks within the PU matrix and the resultant composite material properties were examined as a function of CP loading. Increased PEDOT:PSS loading was found to result in a more connected network within the PU matrix, resulting in increased conductivity and charge storage capacity. Increased CP loading was also determined to increase the Young's modulus and reduce the strain at failure. Biological assessment of CE composites showed them to mediate ReNcell VM human neural precursor cell adhesion. The increased stiffness of CE films was also found to promote neurite outgrowth. CE sheets were directly laser micromachined into a functional array and shown to deliver biphasic waveforms with comparable voltage transients to Pt arrays in in vitro testing.

Conductive elastomer composites for fully polymeric, flexible bioelectronics.

Elastic and conductive hydrogel electrodes.

Intracortical electrodes for brain-machine interfaces rely on intimate contact with tissues for recording signals and stimulating neurons. However, the long-term viability of intracortical electrodes  is poor, with a major contributing factor being the development of a glial scar.  approaches for evaluating responses to intracortical devices are resource intensive and complex, making statistically significant, high throughput data difficult to obtain.  models provide an alternative to  studies; however, existing approaches have limitations which restrict the translation of the cellular reactions to the implant scenario. Notably, there is no current robust model that includes astrocytes, microglia, oligodendrocytes and neurons, the four principle cell types, critical to the health, function and wound responses of the central nervous system (CNS). In previous research a co-culture of primary mouse mature mixed glial cells and immature neural precursor cells were shown to mimic several key properties of the CNS response to implanted electrode materials. However, the method was not robust and took up to 63 days, significantly affecting reproducibility and widespread use for assessing brain-material interactions. In the current research a new co-culture approach has been developed and evaluated using immunocytochemistry and quantitative polymerase chain reaction (qPCR). The resulting method reduced the time in culture significantly and the culture model was shown to have a genetic signature similar to that of healthy adult mouse brain. This new robust CNS culture model has the potential to significantly improve the capacity to translate  data to the  responses.

An Improved  Model of Cortical Tissue.

Hydrogels have been applied across a wide range of biomedical applications due to their versatility, but more recently have garnered interest as materials in bioelectronics due to the capacity to tailor their mechanical and biological properties. Hydrogel coatings in particular have been used to impart softness at the bionic device interface, deliver therapeutics and control cell interactions through presentation of peptides and growth factors. Additionally, the use of dynamic hydrogel properties has been harnessed as shuttles for the implantation of flexible electrode arrays. In all of these applications, the hydrogel must be designed not only to provide the desired performance, but also have no unexpected impacts on the surrounding tissues, such as extensive swelling that can compress the cells at the interface. Appropriate selection and design of hydrogel systems for bioelectronics requires an understanding of the physical, chemical and biological properties of hydrogels as well as their structure-property relationships. This review covers the design rationale for application of hydrogels systems for use in bioelectronic devices with a focus on in vivo applications.

Considerations for hydrogel applications to neural bioelectronics.

Hydrogel materials have been employed as biological scaffolds for tissue regeneration across a wide range of applications. Their versatility and biomimetic properties make them an optimal choice for treating the complex and delicate milieu of neural tissue damage. Aside from finely tailored hydrogel properties, which aim to mimic healthy physiological tissue, a minimally invasive delivery method is essential to prevent off-target and surgery-related complications. The specific class of injectable hydrogels termed self-assembling peptides (SAPs), provide an ideal combination of in situ polymerization combined with versatility for biofunctionlization, tunable physicochemical properties, and high cytocompatibility. This review identifies design criteria for neural scaffolds based upon key cellular interactions with the neural extracellular matrix (ECM), with emphasis on aspects that are reproducible in a biomaterial environment. Examples of the most recent SAPs and modification methods are presented, with a focus on biological, mechanical, and topographical cues. Furthermore, SAP electrical properties and methods to provide appropriate electrical and electrochemical cues are widely discussed, in light of the endogenous electrical activity of neural tissue as well as the clinical effectiveness of stimulation treatments. Recent applications of SAP materials in neural repair and electrical stimulation therapies are highlighted, identifying research gaps in the field of hydrogels for neural regeneration.

Self-Assembling Hydrogel Structures for Neural Tissue Repair.

Conducting polymers (CPs) possess unique electrical and electrochemical properties and hold great potential for different applications in the field of bioelectronics. However, the widespread implementation of CPs in this field has been critically hindered by their poor processibility. There are four key elements that determine the processibility of CPs, which are thermal tunability, chemical stability, solvent compatibility and mechanical robustness. Recent research efforts have focused on enhancing the processibility of these materials through pre- or post-synthesis chemical modifications, the fabrication of CP-based complexes and composites, and the adoption of additive manufacturing techniques. In this review, the physicochemical and structural properties that underlie the performance and processibility of CPs are examined. In addition, current research efforts to overcome technical limitations and broaden the potential applications of CPs in bioelectronics are discussed. STATEMENT OF SIGNIFICANCE: This review details the inherent properties of CPs that have hindered their use in additive manufacturing for the creation of 3D bioelectronics. A fundamental approach is presented with consideration of the chemical structure and how this contributes to their electrical, thermal and mechanical properties. The review then considers how manipulation of these properties has been addressed in the literature including areas where improvements can be made. Finally, the review details the use of CPs in additive manufacturing and the future scope for the use of CPs and their composites in the development of 3D bioelectronics.

The influence of physicochemical properties on the processibility of conducting polymers: A bioelectronics perspective.

Brain-computer interfaces (BCIs) provide bidirectional communication between the brain and output devices that translate user intent into function. Among the different brain imaging techniques used to operate BCIs, electroencephalography (EEG) constitutes the preferred method of choice, owing to its relative low cost, ease of use, high temporal resolution, and noninvasiveness. In recent years, significant progress in wearable technologies and computational intelligence has greatly enhanced the performance and capabilities of EEG-based BCIs (eBCIs) and propelled their migration out of the laboratory and into real-world environments. This rapid translation constitutes a paradigm shift in human-machine interaction that will deeply transform different industries in the near future, including healthcare and wellbeing, entertainment, security, education, and marketing. In this contribution, the state-of-the-art in wearable biosensing is reviewed, focusing on the development of novel electrode interfaces for long term and noninvasive EEG monitoring. Commercially available EEG platforms are surveyed, and a comparative analysis is presented based on the benefits and limitations they provide for eBCI development. Emerging applications in neuroscientific research and future trends related to the widespread implementation of eBCIs for medical and nonmedical uses are discussed. Finally, a commentary on the ethical, social, and legal concerns associated with this increasingly ubiquitous technology is provided, as well as general recommendations to address key issues related to mainstream consumer adoption.

Mind the gap: State-of-the-art technologies and applications for EEG-based brain-computer interfaces.

Neural interfaces (NIs) have traditionally used inorganic device constructs paired with electrical stimulation to bypass injured or diseased electroactive tissues. These bioinert devices have significant impact on the neural tissue, being synthetic and causing large volumetric changes to the biological environment. The concept of biomimicry has become popular for tissue engineering technologies, reflecting biological properties as a component of material design. Tissue engineering strategies can be harnessed in bioelectronic device design to improve biological tolerance, but the need for improved integration with the native tissue remains an unmet need. Adaptive biomimetic designs that respond to the changing neural tissue environment associated with wound healing can actively address the immune response to improve biointegration. These adaptive approaches include responsive materials paired with stem cells and bioactive molecules as integrated components of NIs. Combining adaptive biomimetics with NIs provides a new, more natural approach for communicating with the nervous system.

Adaptive biomimicry: design of neural interfaces with enhanced biointegration.

Recent years have led to a rapid increase in the development of neurotechnologies for diagnosis, monitoring, and treatment of conditions with neurological targets. The central driving force has been the need for next-generation devices to treat neural injury and disease, where current pharmaceutical or conventional bioelectronics have been unable to impart sufficient therapeutic effects. The advent of new therapies and advanced technologies has resulted in a reemergence of the concept of superhuman performance. This is a hypothetical possibility that is enabled when bionics are used to augment the neural system and has included the notions of improved cognitive ability and enhancement of hearing and seeing beyond the limitations of a healthy human. It is quite conceivable that a bionic eye could be used for night vision; however, the damage to both the neural system and surrounding tissues in placing such a device is only considered acceptable in the case of a patient that can obtain improvement in quality of life. There are also critical limitations that have hindered clinical translation of high-resolution neural interfaces, despite significant advances in biomaterial and bioelectronics technologies, including the advent of biohybrid devices. Surgical damage and foreign body reactions to such devices can be reduced but not eliminated, and these engineering solutions to reduce inflammation present additional challenges to the long-term performance and medical regulation. As a result, while bioelectronics has seen concepts from science fiction realized, there remains a significant gap to their use as enhancements beyond medical therapies.

Rylie A. Green

Department of Bioengineering

Imperial College London

Preparation of Rat Sciatic Nerve for Ex Vivo Neurophysiology