Poly(vinyl alcohol)/poly(acrylic acid) hydrogel coatings for improving electrode–neural tissue interface

A major problem which hinders the applications of neural prostheses is the inconsistent performance caused by tissue responses during long-term implantation. The study investigated a new approach for improving the electrode–neural tissue interface. Hydrogel poly(vinyl alcohol)/poly(acrylic acid) interpenetrating polymer networks (PVA/PAA IPNs) were synthesized and tailored as coatings for poly(dimethylsiloxane) (PDMS) based neural electrodes with the aid of plasma pretreatment. Changes in the electrochemical impedance and maximum charge injection (Q_inj) limits of the coated iridium oxide microelectrodes were negligible. Protein adsorption on PDMS was reduced by ～85% after coating. In the presence of nerve growth factor (NGF), neurite extension of rat pheochromocytoma (PC12) cells was clearly greater on PVA/PAA IPN films than on PDMS substrates. Furthermore, the tissue responses of PDMS implants coated with PVA/PAA IPN films were studied by 6-week implantation in the cortex of rats, which found that the glial fibrillary acidic protein (GFAP) immunoreactivity in animals (n = 8) receiving coated implants was significantly lower (p < 0.05) compared to that of uncoated implants (n = 7) along the entire distance of 150 μm from the outer skirt to the implant interface. The coated film remained on the surface of the explanted implants, confirmed by scanning electron microscopy (SEM). All of these suggest the hydrogel coating is feasible and favorable to neural electrode applications.     

Conducting polymers on hydrogel-coated neural electrode provide sensitive neural recordings in auditory cortex

Recently, a significant amount of effort has been dedicated to understanding factors that influence the functionality of bio-electronic sensors and to development of novel coating technologies for modifying biosensor surfaces. Due to its well-known biocompatibility, alginate hydrogel (HG) has been used as a coating material on neural electrodes for promoting intimate cellular integration, providing a scaffold for local drug delivery, and creating a mechanical buffer between hard electrodes and the soft tissues of the central nervous system. However, neural signal recordings using HG-coated electrodes in animal models are still poorly evaluated. Here, we investigated the effect of the proximity of source neurons around the electrode sites using HG coatings with various thicknesses deposited on microfabricated electrodes, implanted in auditory cortex of guinea pigs. We also evaluated the role of the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) in improving the recording functionality of the HG-coated neural electrodes. A significant loss in recording functionality was observed with thicker HG coatings, as determined by the number of clearly detectable units (30% with 80 μm thick coatings) and average signal-to-noise ratios (3.91 with 80 μm thick coatings). However, deposition of the conducting polymer PEDOT on the electrode sites restored the lost functionality of the electrodes caused by the HG coatings (30 μm). These conducting polymer/HG coatings have the potential to improve long-term performance of the neural electrodes not only by improving the electrode biocompatibility but also by facilitating more efficient signal transmission.     

In vivo effects of L1 coating on inflammation and neuronal health at the electrode-tissue interface in rat spinal cord and dorsal root ganglion

The spinal cord (SC) and dorsal root ganglion (DRG) are target implantation regions for neural prosthetics, but the tissue-electrode interface in these regions is not well-studied. To improve understanding of these locations, the tissue reactions around implanted electrodes were characterized. L1, an adhesion molecule shown to maintain neuronal density and reduce gliosis in brain tissue, was then evaluated in SC and DRG implants. Following L1 immobilization onto neural electrodes, the bioactivities of the coatings were verified in vitro using neuron, astrocyte and microglia cultures. Non-modified and L1-coated electrodes were implanted into adult rats for 1 or 4weeks. Hematoxylin and eosin staining along with cell-type specific antibodies were used to characterize the tissue response. In the SC and DRG, cells aggregated at the electrode-tissue interface. Microglia staining was more intense around the implant site and decreased with distance from the interface. Neurofilament staining in both locations decreased or was absent around the implant, compared with surrounding tissue. With L1, neurofilament staining was significantly increased while neuronal cell death decreased. These results indicate that L1-modified electrodes may result in an improved chronic neural interface and will be evaluated in recording and stimulation studies.     

Polylysine-Modified PEG-Based Hydrogels to Enhance the Neuro–Electrode Interface

Neural prostheses are a promising technology in the treatment of lost neural function. However, poor biocompatibility of these devices inhibits the formation of a robust neuro–electrode interface. Several factors including mechanical mismatch between the device and tissue, inflammation at the implantation site, and possible electrical damage contribute to this response. Many researchers are investigating polymeric brain mimetic coatings as a means to improve integration with nervous tissue. Specifically, hydrogels, constructs also employed in tissue engineering, have been explored because of their structural and mechanical similarity to native tissue. However, many hydrogel materials (e.g., poly(ethylene glycol) (PEG)) do not support cell adhesion. In this work, we report a technique to enhance the interface between polymeric brain mimetic coatings and neural tissue using adhesion molecules. In particular, polylysine-modified PEG-based hydrogels were synthesized, characterized and shown to promote neural adhesion using a PC12 cell line. In addition, we examined adhesion behavior of a PEG-co-polymer and found that these materials adhere to electrodes for at least 4 weeks. These results suggest that polylysine–PEG hydrogel biomaterials are biocompatible and can enhance stability of chronic neural interfaces.     

Conducting polymers for neural interfaces: challenges in developing an effective long-term implant

Metal electrode materials used in active implantable devices are often associated with poor long-term stimulation and recording performance. Modification of these materials with conducting polymer coatings has been suggested as an approach for improving the neural tissue-electrode interface and increasing the effective lifetime of these implants. Neural interfaces ideally have intimate contact between the excitable tissue and the electrode to maintain signal quality and activation of neural cells. The outcomes of current research into conducting polymers as coatings has potential to enhance this tissue-material contact by increasing the electrode surface area and roughness as well as allowing delivery of bioactive signals to neural cells. However, challenges facing conducting polymers include poor electroactive stability and mechanical properties as well as control of the mobility, concentration and presentation of bioactive molecules. The impact of biological inclusions on polymer properties and their ongoing performance in neural prosthetics requires a greater understanding with future research aimed at controlling and optimising film characteristics for long-term performance. Optimising the electrode interface will require a trade-off between desired electrical, mechanical, chemical and biological properties.     

神经电极的水凝胶涂层及楔形角对组织损伤的影响

目的研究水凝胶涂层的厚度以及电极楔形角对植入损伤的影响。方法基于植入损伤评估系统,进行模拟神经电极植入过程的实验,评估电极植入造成的组织损伤。用浸涂次数(分别为0、1、2、3)控制水凝胶涂层的厚度,选用30°、40°、50°、60°为电极楔形角的变量。以最大组织应变和最大植入力为组织损伤的衡量标准。结果水凝胶涂层越厚,电极植入损伤越大。楔形角越小,植入损伤也越小。同时,减小针尖的楔形角可以减小涂层对组织损伤的影响程度。3次浸涂时,楔形角为30°时,最大组织应变与最大植入力分别增加3.4%和3.8%,而楔形角为60°时,两者分别增加11.3%和18.1%。结论神经电极的水凝胶涂层将增大植入电极对生物组织的损伤,然而减小电极尖端楔形角的方法可以降低水凝胶涂层厚度对植入损伤的影响程度。     

A strategy to passively reduce neuroinflammation surrounding devices implanted chronically in brain tissue by manipulating device surface permeability

Available evidence indicates that pro-inflammatory cytokines produced by immune cells are likely responsible for the negative sequela associated with the foreign body response (FBR) to chronic indwelling implants in brain tissue. In this study a computational modeling approach was used to design a diffusion sink placed at the device surface that would retain pro-inflammatory cytokines for sufficient time to passively antagonize their impact on the FBR. Using quantitative immunohistochemistry, we examined the FBR to such engineered devices after a 16-week implantation period in the cortex of adult male Sprague–Dawley rats. Our results indicate that thick permeable surface coatings, which served as diffusion sinks, significantly reduced the FBR compared to implants either with no coating or with a thinner coating. The results suggest that increasing surface permeability of solid implanted devices to create a diffusion sink can be used to reduce the FBR and improve biocompatibility of chronic indwelling devices in brain tissue.     

Localized controlled release of stratifin reduces implantation-induced dermal fibrosis

Localized controlled release of anti-fibrogenic factors can potentially prevent tissue fibrosis surrounding biomedical prostheses, such as vascular stents and breast implants. We have previously demonstrated that therapeutic intervention with topically applied stratifin in a rabbit ear fibrotic model not only prevents dermal fibrosis but also promotes more normal tissue repair by regulating extracellular matrix deposition. In this work, the anti-fibrogenic effect of a controlled release form of stratifin was investigated in the prevention of fibrosis induced by dermal poly(lactic-co-glycolic acid) (PLGA) microsphere/poly(vinyl alcohol) (PVA) hydrogel implants. Pharmacodynamic effects were evaluated by histopathological examination of subcutaneous tissue surrounding implanted composites. Controlled release of stratifin from PLGA microsphere/PVA hydrogel implants significantly moderated dermal fibrosis and inflammation by reducing collagen deposition (30%), total tissue cellularity (48%) and infiltrated CD3(+) immune cells (81%) in the surrounding tissue compared with the stratifin-free implants. The controlled release of stratifin from implants markedly increased the level of matrix metalloproteinase-1 expression in the surrounding tissue, which resulted in less collagen deposition. These stratifin-eluting PLGA/PVA composites show promise as coatings to decrease the typical fibrosis exhibited around implanted biomedical prostheses, such as breast implants and vascular stents.     

Surface modification of neural recording electrodes with conducting polymer/biomolecule blends

The interface between micromachined neural microelectrodes and neural tissue plays an important role in chronic in vivo recording. Electrochemical polymerization was used to optimize the surface of the metal electrode sites. Electrically conductive polymers (polypyrrole) combined with biomolecules having cell adhesion functionality were deposited with great precision onto microelectrode sites of neural probes. The biomolecules used were a silk-like polymer having fibronectin fragments (SLPF) and nonapeptide CDPGYIGSR. The existence of protein polymers and peptides in the coatings was confirmed by reflective microfocusing Fourier transform infrared spectroscopy (FTIR). The morphology of the coating was rough and fuzzy, providing a high density of bioactive sites for interaction with neural cells. This high interfacial area also helped to lower the impedance of the electrode site and, consequently, to improve the signal transport. Impedance spectroscopy showed a lowered magnitude and phase of impedance around the biologically relevant frequency of 1 kHz. Cyclic voltammetry demonstrated the intrinsic redox reaction of the doped polypyrrole and the increased charge capacity of the coated electrodes. Rat glial cells and human neuroblastoma cells were seeded and cultured on neural probes with coated and uncoated electrodes. Glial cells appeared to attach better to polypyrrole/SLPF-coated electrodes than to uncoated gold electrodes. Neuroblastoma cells grew preferentially on and around the polypyrrole/CDPGYIGSR-coated electrode sites while the polypyrrole/CH(3)COO(-)-coated sites on the same probe did not show a preferential attraction to the cells. These results indicate that we can adjust the chemical composition, morphology, electronic transport, and bioactivity of polymer coatings on electrode surfaces on a multichannel micromachined neural probe by controlling electrochemical deposition conditions.     

Glial cell and fibroblast cytotoxicity study on plasma-deposited diamond-like carbon coatings

Diamond-like carbon films have been evaluated as coatings to improve biocompatibility of orthopedic and cardiovascular implants. This study initiates a series of investigations that will evaluate diamond-like carbon (DLC) as a coating for improved biocompatibility in chronic neuroprosthetic implants. Studies in this report assess the cytotoxicity and cell adhesion behavior of DLC coatings exposed to glial and fibroblast cell lines in vitro. It can be concluded from these studies that DLC coatings do not adversely affect 3T3 fibroblast and T98-G glial cell function in vitro. We also successfully rendered DLC coatings non-adhesive (no significant fibroblast or glial cell adhesion) with surface immobilized dextran using methods developed for other biomaterials and applications. Future work will further develop DLC coatings on prototype microelectrode devices for chronic neural implant applications.     

Poly(3,4-ethylenedioxythiophene)/multiwall carbon nanotube composite coatings for improving the stability of microelectrodes in neural prostheses applications

With the purpose of improving the stability of microelectrodes under continuous high charge density stimulation, which is required for neural prostheses applications such as visual prostheses, multiwall carbon nanotube (MWCNT)-doped poly(3,4-ethylenedioxythiophene) (PEDOT) composite films were coated onto a platinum microelectrode by electrochemical polymerization. Galvanostatically polymerized PEDOT/MWCNT films demonstrated superior characteristics compared to polystyrene sulfonate doping and potentiostatic polymerization, including a three-dimensional cone morphology and enhanced electrochemical performance (the safe charge injection limit reached 6.2 mC cm^?2 for cathodic-first pulses). Most important of all, the improved stability of the coatings has been revealed through stimulation for 96 h using 3.0 mC cm^?2 current pulses in bicarbonate- and phosphate-buffered saline solution. Cell assays revealed that PEDOT/MWCNT films could promote the adhesion and neurite outgrowth of rat pheochromocytoma cells. Finally, platinum wires coated with PEDOT/MWCNT films were implanted into rat cortex for 6 weeks for histological evaluation. Glial fibrillary acidic protein and neuronal nuclei staining revealed that the films elicit a lower tissue response compared to platinum implants. These results suggest that the galvanostatically polymerized PEDOT/MWCNT films can improve the stability of stimulation microelectrodes and that PEDOT/MWCNT is an excellent candidate material for electrode coating for neural prostheses applications.     

In vivo stability and biocompatibility of implanted calcium alginate disks

Alginate is a commonly used biomedical hydrogel whose in vivo degradation behavior is only beginning to be understood. The use of alginate in the central nervous system is gaining popularity as an electrode coating, cell encapsulation matrix, and for duraplasty. However, it is necessary to understand how the hydrogel will behave in vivo to aid in the development of alginate for use as a neural interface material. The goal of the current study was to compare the rheological behavior of explanted alginate disks and the inflammatory response to subcutaneously implanted alginate hydrogels over a 3-month period. Specifically, the effects due to (1) in situ gelling, (2) diffusion gelling, and (3) use of a poly-l-lysine (PLL) coating were investigated. While all samples' complex moduli decreased 80% in the first day, in situ gelled alginate was more stable for the first week of implantation. The PLL coating offered some stability increases for diffusion gelled alginate, but the stability in both conditions remained significantly lower than that in in situ gelled alginate. There were no differences in biocompatibility that clearly suggested one gelation method over another. These results indicate that in situ gelation is the preferred method in neural interface applications where stability is the primary concern.     

In vivo dissolution behavior of various RF magnetron-sputtered Ca-P coatings on roughened titanium implants

RF magnetron sputter deposition was used to produce 0.1, 1.0 and 4.0 microm thick Ca-P coatings on TiO(2)-blasted titanium discs. Half of the as-sputtered coated specimens were subjected to an additional infrared heat treatment for 30s at 425-475 degrees C. X-ray diffraction demonstrated that infrared radiation changed the amorphous 4 microm sputtered coatings into an amorphous-crystalline structure, while the amorphous 0.1 and 1 microm changed in a crystalline apatite structure with the presents of tetracalciumphosphate as a second phase. Scanning electron microscopically examination of the sputtered coatings revealed that annealing of the 4 microm thick coatings resulted in the appearance of small cracks. Subsequently, the discs were implanted subcutaneous into the back of rabbits. After 1, 4, 8 and 12 weeks of implantation, the implants were retrieved and prepared for histological and physicochemical evaluation. Histological evaluation revealed that the tissue response to all coated implants was very uniform. A very thin connective tissue capsule surrounded all implants. The capsule was usually free of inflammatory cells. At the interface, there was a close contact between the capsule and implant surface and no inflammatory cells were seen. Physicochemical evaluation showed that the 0.1 and 1 microm thick amorphous coatings had disappeared within 1 week of implantation. On the other hand, the 4 microm thick amorphous phase disappeared during the implantation periods, which was followed by the precipitation of a crystalline carbonate apatite. Further, at all implantation periods the heat-treated 1 and 4 microm thick coatings could be detected. Occasionally, a granular precipitate was deposited on the heat-treated 4 microm thick coating. Fourier transform infrared spectroscopy showed the formation of carbonate apatite (CO(3)-AP) on the 4 microm thick amorphous coating and on the heat-treated specimens. On basis of our findings, we conclude that 1 microm thick heat-treated Ca-P sputter coating on roughened titanium implants appear to be of sufficient thickness to show bioactive properties, under in vivo conditions.     

Reduced foreign body response at nitric oxide-releasing subcutaneous implants

The tissue response to nitric oxide (NO)-releasing subcutaneous implants is presented. Model implants were created by coating silicone elastomer with diazeniumdiolate-modified xerogel polymers capable of releasing NO. The host tissue response to such implants was evaluated at 1, 3, and 6 weeks and compared to that of uncoated silicone elastomer blanks and xerogel-coated controls incapable of releasing NO. Delivery of NO (approximately 1.35 micromol/cm2 of implant surface area) reduced foreign body collagen capsule ("scar tissue") thickness by >50% compared to uncoated silicone elastomer after 3 weeks. The chronic inflammatory response at the tissue/implant interface was also reduced by >30% at NO-releasing implants after 3 and 6 weeks. Additionally, CD-31 immunohistochemical staining revealed approximately 77% more blood vessels in proximity to NO-releasing implants after 1 week compared to controls. These findings suggest that conferring NO release to subcutaneous implants may promote effective device integration into healthy vascularized tissue, diminish foreign body capsule formation, and improve the performance of indwelling medical devices that require constant mass transport of analytes (e.g., implantable sensors).     

Permeability of subcutaneous tissues surrounding long-term implants to oxygen

Certain types of implanted medical devices depend on oxygen supplied from surrounding tissues for their function. However, there is a concern that the tissue associated with the foreign body response to implants may become impermeable to oxygen over the long term and render the implant nonfunctional. We report oxygen flux recordings from electrochemical oxygen sensor devices with wireless telemetry implanted in subcutaneous porcine tissues. The devices remained implanted for up to 13 weeks and were removed with adjacent tissues at specified times for histologic examination. There are four main observations: (1) In the first few weeks after implantation, the oxygen flux to the sensors, or current density, declined to a sustained mean value, having unsynchronized cyclic variations around the mean; (2) The oxygen mass transfer resistance of the sensor membrane was negligible compared to that of the tissue, allowing for a sensitive estimate of the tissue permeability; (3) The effective diffusion coefficient of oxygen in tissues was found to be approximately one order of magnitude lower than in water; and (4) Quantitative histologic analysis of the tissues showed a mild foreign body response to the PDMS sensor membrane material, with capillaries positioned close to the implant surface. Continuous recordings of oxygen flux indicate that the tissue permeability changes predictably with time, and suggest that oxygen delivery can be sustained over the long term.     

Characterization of microglial attachment and cytokine release on biomaterials of differing surface chemistry

The clinical usefulness of central nervous system recording electrodes is currently limited by inconsistent long-term performance that is believed to be governed by the brain tissue response to the implant. In this study, we observed persistent macrophage biomarker expression at the biotic-abiotic interface surrounding implanted electrodes over a 12-week indwelling period. Using the cell type-specific marker CD11b to examine the cells attached to electrodes retrieved over the indwelling period, we found that most of the cells were activated microglia, the resident macrophage of brain tissue, indicating that the implanted electrodes behave as a persistent inflammatory stimulus. To determine the potential usefulness of different materials as coatings for implanted electrodes, we examined brain-derived microglial cell attachment and cytokine release on a number of medically relevant materials. Our results suggest that activated microglia attach to many of the materials used as external coatings for electrode manufacture, and likely serve as a source of pro-inflammatory and neurotoxic cytokines that may be responsible for reducing the biocompatibility of such implants. Our results also indicate that low protein-binding coatings may be useful in reducing microglial attachment upon implantation in brain tissue and may provide a means of improving electrode biocompatibility.     

Long term performance of porous platinum coated neural electrodes

Over the last several years, there has been a growing interest in neural implants for the study and diagnostics of neurological disorders as well as for the symptomatic treatment of central nervous system related diseases. One of the major challenges is the trade-off between small electrode sizes for high selectivity between single neurons and large electrode-tissue interface areas for excellent stimulation and recording properties. This paper presents an approach of increasing the real surface area of the electrodes by creating a surface microstructure. Two major novelties let this work stand out from existing approaches which mainly make use of porous coatings such as platinum black or iridium oxide, or Poly(3,4-ethylenedioxythiophene) (PEDOT). Roughening is carried out by a dry etching process on the silicon electrode core before being coated by a sputtered platinum layer, eliminating complicated deposition processes as for the materials described above. The technology is compatible with any commonly used coating material. In addition, the surface roughening is compatible with high aspect ratio penetrating electrode arrays such as the well-established Utah electrode array, whose unique geometry presents a challenge in the surface modification of active electrode sites. The dry etching process is well characterized and yields a high controllability of pore size and depth. This paper confirms the superior electrochemical properties including impedance, charge injection capacity, and charge storage capacity of surface engineered electrode arrays compared to conventional arrays over a period of 12 weeks. Furthermore, mechanical stability of the modified electrodes was tested by implantation in the brain of a recently deceased rat. In conclusion, the larger interface surface of the electrodes does not only decrease the impedance which should lead to enhanced Signal to noise ratio (SNR) for recording purposes, but also yields higher charge injection capacities, which improve the stimulation characteristics of the implants.     

Bone bonding to hydroxyapatite and titanium surfaces on femoral stems retrieved from human subjects at autopsy

The success of clinical results obtained with many hydroxyapatite (HA)-coated prosthetic designs has deflected attention from the need to extend the life of the HA coating on the device. In the current study the percentages of HA and titanium surfaces to which bone was bonded, on HA-coated and non-coated titanium femoral stems retrieved from human subjects, were evaluated. Plasma-sprayed hydroxyapatite (PSHA)-coated devices demonstrated wide variability in the percentage of the PSHA coating remaining on the stems. The coating was missing from a substantial portion of a stem after only about 6 months of implantation. The percentage of revealed metal to which bone was bonded was significantly less than the percentage of the HA coating demonstrating such bonding. The revealed metal to which bone was bonded was comparable to the same value for a separate group of non-PSHA-coated titanium stems. If HA-coatings degrade over time precipitous decline in performance may occur even after several functional years. Many ultrastructural features of the bone bonded to the HA coatings on these implants from human subjects were comparable to those found on HA-coated devices implanted in a canine model.     

The surface immobilization of the neural adhesion molecule L1 on neural probes and its effect on neuronal density and gliosis at the probe/tissue interface

Brain tissue inflammatory responses, including neuronal loss and gliosis at the neural electrode/tissue interface, limit the recording stability and longevity of neural probes. The neural adhesion molecule L1 specifically promotes neurite outgrowth and neuronal survival. In this study, we covalently immobilized L1 on the surface of silicon-based neural probes and compared the tissue response between L1 modified and non-modified probes implanted in the rat cortex after 1, 4, and 8 weeks. The effect of L1 on neuronal health and survival, and glial cell reactions were evaluated with immunohistochemistry and quantitative image analysis. Similar to previous findings, persistent glial activation and significant decreases of neuronal and axonal densities were found at the vicinity of the non-modified probes. In contrast, the immediate area (100 μm) around the L1 modified probe showed no loss of neuronal bodies and a significantly increased axonal density relative to background. In this same region, immunohistochemistry analyses show a significantly lower activation of microglia and reaction of astrocytes around the L1 modified probes when compared to the control probes. These improvements in tissue reaction induced by the L1 coating are likely to lead to improved functionality of the implanted neural electrodes during chronic recordings.     

Reduced acute inflammatory responses to microgel conformal coatings

Implantation of synthetic materials into the body elicits inflammatory host responses that limit medical device integration and biological performance. This inflammatory cascade involves protein adsorption, leukocyte recruitment and activation, cytokine release, and fibrous encapsulation of the implant. We present a coating strategy based on thin films of poly(N-isopropylacrylamide) hydrogel microparticles (i.e. microgels) cross-linked with poly(ethylene glycol) diacrylate. These particles were grafted onto a clinically relevant polymeric material to generate conformal coatings that significantly reduced in vitro fibrinogen adsorption and primary human monocyte/macrophage adhesion and spreading. These microgel coatings also reduced leukocyte adhesion and expression of pro-inflammatory cytokines (TNF-alpha, IL-1beta, MCP-1) in response to materials implanted acutely in the murine intraperitoneal space. These microgel coatings can be applied to biomedical implants as a protective coating to attenuate biofouling, leukocyte adhesion and activation, and adverse host responses for biomedical and biotechnological applications.