An in vitro model of a retinal prosthesis

Epiretinal prostheses are being developed to bypass a degenerated photoreceptor layer and excite surviving ganglion and inner retinal cells. We used custom microfabricated multielectrode arrays with 200-microm-diameter stimulating electrodes and 10-microm-diameter recording electrodes to stimulate and record neural responses in isolated tiger salamander retina. Pharmacological agents were used to isolate direct excitation of ganglion cells from excitation of other inner retinal cells. Strength-duration data suggest that, if amplitude will be used for the coding of brightness or gray level in retinal prostheses, shorter pulses (200 micros) will allow for a smaller region in the area of the electrode to be excited over a larger dynamic range compared with longer pulses (1 ms). Both electrophysiological results and electrostatic finite-element modeling show that electrode-electrode interactions can lead to increased thresholds for sites half way between simultaneously stimulated electrodes (29.4 +/- 6.6 nC) compared with monopolar stimulation (13.3 +/- 1.7 nC, p < 0.02). Presynaptic stimulation of the same ganglion cell with both 200- and 10-microm-diameter electrodes yielded threshold charge densities of 12 +/- 6 and 7.66 +/- 1.30 nC/cm2, respectively, while the required charge was 12.5 +/- 6.2 and 19 +/- 3.3 nC.     

An In Vitro Model of a Retinal Prosthesis

Epiretinal prostheses are being developed to bypass a degenerated photoreceptor layer and excite surviving ganglion and inner retinal cells. We used custom microfabricated multielectrode arrays with 200-mum-diameter stimulating electrodes and 10-mum-diameter recording electrodes to stimulate and record neural responses in isolated tiger salamander retina. Pharmacological agents were used to isolate direct excitation of ganglion cells from excitation of other inner retinal cells. Strength-duration data suggest that, if amplitude will be used for the coding of brightness or gray level in retinal prostheses, shorter pulses (200 mus) will allow for a smaller region in the area of the electrode to be excited over a larger dynamic range compared with longer pulses (1 ms). Both electrophysiological results and electrostatic finite-element modeling show that electrode-electrode interactions can lead to increased thresholds for sites half way between simultaneously stimulated electrodes (29.4 plusmn 6.6 nC) compared with monopolar stimulation (13.3 plusmn 1.7 nC, < 0.02). Presynaptic stimulation of the same ganglion cell with both 200- and 10- m-diameter electrodes yielded threshold charge densities of 12 plusmn 6 and 7.66 plusmn 1.30 nC/cm², respectively, while the required charge was 12.5 plusmn 6.2 and 19 plusmn 3.3 nC.     

A method for evaluating the selectivity of electrodes implanted fornerve simulation

The scale of stimulating electrodes possible for use in functional electrical stimulation to restore motor and sensory function is rapidly approaching that of individual neurons. Although the electrodes may approach the dimensions of single nerve cells, it is unclear if the region of excitation elicited by each electrode will be correspondingly small. Previous techniques for evaluating this have either been tedious or have lacked the resolution necessary. This paper describes a method that uses the refractory interaction of the compound action potentials elicited by a stimulus pulse pair, along with high-resolution recording of those potentials, to achieve measurements of the selectivity of stimulation down to the scale of a few axon diameters. The feasibility of this technique is demonstrated in sciatic nerves of frogs (Rana Catesbiana) acutely implanted with a sapphire electrode array.     

A model of safe levels for electrical stimulation

A model is presented that represents a large body of data on safety and damage levels of electrical stimulation. The predictions of the model are consistent with known principles of current flow and known mechanisms of damage around stimulating electrodes. It is proposed that limits on levels of electrical stimulation take into account the location of the electrode relative to the stimulated tissue and these limits can be computed algorithmically from the model.     

Tissue damage by pulsed electrical stimulation

Repeated pulsed electrical stimulation is used in a multitude of neural interfaces; damage resulting from such stimulation was studied as a function of pulse duration, electrode size, and number of pulses using a fluorescent assay on chick chorioallontoic membrane (CAM) in vivo and chick retina in vitro. Data from the chick model were verified by repeating some measurements on porcine retina in-vitro. The electrode size varied from 100 microm to 1 mm, pulse duration from 6 micros to 6 ms, and the number of pulses from 1 to 7500. The threshold current density for damage was independent of electrode size for diameters greater than 300 microm, and scaled as 1/r2 for electrodes smaller than 200 microm. Damage threshold decreased with the number of pulses, dropping by a factor of 14 on the CAM and 7 on the retina as the number of pulses increased from 1 to 50, and remained constant for a higher numbers of pulses. The damage threshold current density on large electrodes scaled with pulse duration as approximately 1/t0.5, characteristic of electroporation. The threshold current density for repeated exposure on the retina varied between 0.061 A/cm2 at 6 ms to 1.3 A/cm2 at 6 micros. The highest ratio of the damage threshold to the stimulation threshold in retinal ganglion cells occurred at pulse durations near chronaxie-around 1.3 ms.     

The effect of anisotropy on the potential distribution in biological tissue and its impact on nerve excitation simulations

We present a finite difference solution of the potential distribution associated with electrical current stimulation in an anisotropic in-homogeneous tissue environment and compare it to the isotropic case. The results demonstrate that there can be significant errors associated with the assumption of isotropic tissue properties in calculating the potential distribution along an axon in nerve excitation simulations. These errors can have a significant impact on predicted nerve fiber recruitment patterns when evaluating the efficacy of specific surface or intramuscular stimulus electrode configurations. The results of this study also suggest when a more comprehensive tissue model should be implemented in an electrode design study. Simulation results indicate that the isotropy assumption is worst under bipolar electrode stimulation as opposed to monopolar stimulation and that the bipolar error increases as the distance between electrodes decreases. In light of these results, it is concluded that in order to avoid large errors in the calculated potential distribution along an axon, the isotropy assumption should only be used when the transverse depth from the electrode to the nerve is relatively small.     

Localized electrical nerve blocking

Localized electrical nerve blocking was investigated in computer simulation and in vivo trials for sinusoidal frequencies between 5 and 20 kHz. Computer simulations indicated that a localized transmission block of the axons could occur in each of the axon models. An approximation of nerve stimulation was derived from individual axon simulations conducted over axon diameters of 5-15 microm and electrode to axon distances of 0.25 to 2.0 mm. Examination of the membrane voltage and ionic gate potentials indicated that the block could be attributed to an elevated membrane voltage. The elevated membrane voltage could prevent conduction of action potentials through the region of the sinusoidal currents. At lower amplitudes, the sinusoidal current could stimulate the axon and generate a continuous series of action potentials. In vivo trials demonstrated that the sinusoidal frequencies of greater than 10 kHz would cause a localized block in rats. Sinusoidal frequencies below 5 kHz would lead to a reduction in muscle force that appeared to be caused by depletion of transmitter at the neuromuscular junction. As indicated by the computer models of rat nerves, the endplate depletion block occurred at a lower frequency (below 5 kHz) than the block (above 10 kHz). A partial block of the axon was demonstrated, suggesting that sinusoidal currents could be used to provide selective stimulation if they are combined with distal electrical stimulation.     

Maintaining Cytocompatibility of Biopolymers Through a Graphene Layer for Electrical Stimulation of Nerve Cells

Here, the utility of large‐area graphene as a flexible, biocompatible electrode to stimulate cell growth is demonstrated. Chemical vapor deposition allows the production of highly crystalline, single, double, or few‐layered graphene on copper substrates. The subsequent transfer to a biopolymer support, such as polylactic acid (PLA) or polylactic‐co‐glycolic acid (PLGA) copolymers, provides a unique electrode structure retaining the flexibility and surface properties of the underlying materials with a conductive graphene layer sufficient to enable electrical communication with excitable cells. The growth and compatibility of PC‐12 cells on these graphene‐biopolymer (GPB) electrodes is influenced more by the underlying polymer than the presence of graphene, demonstrating that the characteristics influencing biocompatibility have been retained after graphene modification. Differentiation of these cells into neural phenotypes is enhanced using electrical stimulation through the graphene conductive layer, confirming that the conductivity of graphene is sufficient to electrically communicate with cells grown on the surface. The process described herein demonstrates that non‐conducting, flexible biopolymer surfaces can be easily coated with graphene without changing the biocompatibility of the materials. This could be used to create electrodes from non‐conducting materials with optimized cell compatibility with graphene providing electrical properties suitable for stimulation of cells without greatly changing the surface properties.     

Charge density and charge per phase as cofactors in neural injuryinduced by electrical stimulation

The possibility of neural injury during prolonged electrical stimulation of the brain imposes some constraints on the use of this technique for therapeutic and experimental applications. Stimulating electrodes of various sizes were used to investigate the interactions of two stimulus parameters, charge density and charge per phase, in determining the threshold of neural injury induced by electrical stimulation. Platinum electrodes ranging in size from 0.002 to 0.5 cm2 were implanted over the parietal cortex of adult cats. Penetrating microelectrodes fabricated from iridium, with surface areas of 65 +/- 3 x 10(-6) cm2 were inserted into the parietal cortex. Ten days after implantation, the electrodes were pulsed continuously for 7h using charge balanced, current regulated, symmetric pulse pairs, 400 microseconds per phase in duration, at a repetition rate of 50 Hz. The animals were perfused immediately after the stimulation for histologic evaluation of the brain tissue subjacent to the electrode sites. The results show that charge density (as measured at the surface of the stimulating electrode), and charge per phase, interact in a synergistic manner to determine the threshold of stimulation-induced neural injury. This interaction occurs over a wide range of both parameters; for charge density from at least 10 to 800 microC/cm2 and, for charge per phase, from at least 0.05 to 5.0 microC per phase. The significance of these findings in elucidating the mechanisms underlying stimulation-induced injury is discussed.     

The Role of Slow Potassium Current in Nerve Conduction Block Induced by High-Frequency Biphasic Electrical Current

The role of slow potassium current in nerve conduction block induced by high-frequency biphasic electrical current was analyzed using a lumped circuit model of a myelinated axon based on the Schwarz–Reid–Bostock model. The results indicate that nerve conduction block at stimulation frequencies above 4 kHz is due to constant activation of both fast and slow potassium channels, but the block at stimulation frequencies below 4 kHz could be due to either anodal or cathodal dc block depending on the time of the action potiential arriving at the block electrode. When stimulation frequency was above 4 kHz, the slow potassium current was about 3.5 to 6.5 times greater than the fast potassium current at blocking threshold, indicating that the slow potassium current played a more dominant role than the fast potassium current. The blocking location moved from the node under the blocking electrode to a nearby node as the stimulation intensity increased. This simulation study reveals that in mammalian myelinated axons, the slow potassium current probably plays a critical role in the nerve conduction block induced by high-frequency biphasic electrical current.      

Simulation analysis of conduction block in unmyelinated axons induced by high-frequency biphasic electrical currents

Nerve conduction block induced by high-frequency biphasic electrical currents is analyzed using a lumped circuit model of the unmyelinated axon based on Hodgkin-Huxley equations. Axons of different diameters (5-20 microm) can not be blocked completely when the stimulation frequency is between 2 kHz and 4 kHz. However, when the stimulation frequency is above 4 kHz, all axons can be blocked. At high-frequency a higher stimulation intensity is needed to block nerve conduction. The larger diameter axon has a lower threshold intensity for conduction block. The stimulation waveform in which the pulsewidth changes with frequency is more effective in blocking nerve conduction than the waveform in which the pulsewidth is fixed. The activation of potassium channels, rather than inactivation of sodium channels, is the possible mechanism underlying the nerve conduction block of the unmyelinated axon. This simulation study further increases our understanding of axonal conduction block induced by high-frequency biphasic currents, and can guide future animal experiments as well as optimize stimulation waveforms that might be used for electrical nerve block in clinical applications.     

Selectivity of multiple-contact nerve cuff electrodes: a simulation analysis

Advances in functional neuromuscular stimulation (FNS) have increased the need for nerve cuff designs that can control multiple motor functions through selective stimulation of selected populations of axons. This selectivity has proved to be difficult to achieve. Recent experiments suggest that it is possible to slowly reshape peripheral nerve without affecting its physiological function. Using computer simulations we have tested the hypothesis that changing the cross section of a nerve from a round to a flat configuration can significantly improve the selectivity of a nerve cuff. We introduce a new index to estimate selectivity to evaluate the various designs. This index is based on the ability of a nerve electrode to stimulate a target axon without stimulating any other axons. The calculations involve a three-dimensional finite element model to represent the electrical properties of the nerve and cuff and the determination of the firing properties of individual axons. The selectivity rating was found to be significantly higher for the Flat Cuff than the Round Cuff. The result was valid with uniform or random distribution of axons and with a random distribution of fascicles diameters. Flattening of individual fascicles also improved the selectivity of the Flat Cuff but only when the number of contacts used was increased to maintain uniform contact density. Therefore, cuff designs that can reshape the nerve into flatter configurations should yield better cuff performance than the cylindrical cuffs but will require higher contact density.     

Proposal of a new method for narrowing and moving the stimulated region of cochlear implants: animal experiment and numerical analysis

We have proposed the tripolar electrode stimulation method (TESM) for narrowing the stimulation region and continuously moving the stimulation site for cochlear implants. The TESM stimulates the auditory nerve array using three adjacent electrodes which are selected among the electrodes of an electrode array within the lymphatic fluid. Current is emitted from each of the two lateral electrodes and received by the central electrode. The current received by the central electrode is made equal to the sum of the currents emitted from the lateral electrodes. In this paper, we evaluate whether or not TESM works according to a theory which is based on numerical analysis using an electrical equivalent circuit model of the auditory nerve fibers. In this simulation, the sums of the excited model fibers are compared to the compound action potentials (CAP's) which we obtained through animal experiments. To identify the main parameter while maintaining the amplitude of the CAP (the sum of the fired fibers), we assumed the presence of some parameters from the radial current density profile. In the case of the width value among the parameters being kept constant, the amplitude of the CAP was almost constant; thus, the number of the fired fibers was also almost constant. The width value equals the distance between the points at which the profile of the radial current density of the electrode array and the line of the radial threshold current density of the electrode array intersect. It is possible to determine the measure of the stimulation region or site by controlling the width value and the ratios of the currents emitted from the lateral electrodes. As a result, we succeeded in narrowing the stimulation region by controlling the sum of the currents emitted from the two lateral electrodes. Also we succeeded in continuously moving the stimulation site by modifying the currents emitted from the two lateral electrodes.     

In Vivo Electrical Stimulation Using Multichannel Photolithographic Electrode Atrays

Multichannel electrical stimulation of the auditory nerve is demonstrated in a cat model using photolithographic electrode arrays. Evoked potentials from the auditory cortex are used to map the location of fibers activated by different electrodes in the array. The evoked responses obtained are equivalent to those produced by fine wire electrodes currently used for functional stimulation of the auditory system.     

CMOS retinal prosthesis with on-chip electrode impedance measurement

A retinal prosthesis device with built-in self-test capability is proposed and demonstrated. The measurement of electrode impedance as a self-test is achieved without increasing the chip area by employing analogue multiplexers to allow the electrodes to be used for both stimulation of retinal cells and measurement of impedance. Measurement is performed using a four-terminal method to ensure good accuracy. A prototype stimulus chip with 16/spl times/16 channels is fabricated using standard 0.6 /spl mu/m CMOS technology, and is demonstrated to provide self-test functionality with error of as little as 0.05% in the frequency range of 100 Hz to 100 kHz.     

Inversion of the current-distance relationship by transientdepolarization

The objective of this research was to develop a technique to excite selectively nerve fibers distant from an electrode without exciting nerve fibers close to the electrode. The shape of the stimulus current waveform was designed based on the nonlinear conductance properties of neuronal sodium channels. Models of mammalian peripheral myelinated axons and experimental measurements on cat sciatic nerve were used to determine the effects of subthreshold polarization on neural excitability and recruitment. Subthreshold membrane depolarization generated a transient decrease in neural excitability and thus an increase in the threshold for stimulation by a subsequent stimulus pulse. The decrease in excitability increased as the duration and amplitude of the subthreshold depolarization were increased, and the increase in threshold was greater for fibers close to the electrode. When a depolarizing stimulus pulse was applied immediately after the subthreshold depolarization, nerve fibers far from the electrode could be stimulated without stimulating fibers close to the electrode. Subthreshold depolarizing prepulses inverted the current-distance relationship and allowed selective stimulation of nerve fibers far from the electrode     

Effect of surface boundary on neuronal magnetic stimulation

The effect of the surface boundary between free space and a conducting medium on the excitation properties of neurons by magnetic fields are analyzed. The electric field and the spatial derivative of the induced field generated by a coil mounted both parallel and perpendicular to the surface of a semi-infinite conducting medium were calculated using the method of images. An imaginary axon is located in the same relative position from the coil in both configurations and the excitation properties are compared. The calculations are expressed in terms of the activating function for the electrical stimulation of axons. The calculations indicate that the activating function for magnetic stimulation is biphasic as opposed to triphasic for electrical stimulation. The large spatial extent of the magnetically induced electric field compared to the electric field generated by point source electrode suggests a different mode of excitation for neuronal structures in the CNS. The field distribution have been verified experimentally and are important for the understanding of the mechanisms of magnetic stimulation of neural tissue.     

Strategies to improve electrode positioning and safety in cochlear implants

An injection-molded internal supporting rib has been produced to control the flexibility of silicone rubber encapsulated electrodes designed to electrically stimulate the auditory nerve in human subjects with severe to profound hearing loss. The rib molding dies, and molds for silicone rubber encapsulation of the electrode, were designed and machined using AutoCad and MasterCam software packages in a PC environment. After molding, the prototype plastic ribs were iteratively modified based on observations of the performance of the rib/silicone composite insert in a clear plastic model of the human scala tympani cavity. The rib-based electrodes were reliably inserted farther into these models, required less insertion force and were positioned closer to the target auditory neural elements than currently available cochlear implant electrodes. With further design improvements the injection-molded rib may also function to accurately support metal stimulating contacts and wire leads during assembly to significantly increase the manufacturing efficiency of these devices. This method to reliably control the mechanical properties of miniature implantable devices with multiple electrical leads may be valuable in other areas of biomedical device design.     

Mobile Silk Fibroin Electrode for Manipulation and Electrical Stimulation of Adherent Cells

Microfabricated mobile electrodes, nanopallets, designed to provide adherent cells with electrical stimulation are reported. Nanopallets composed of a cross‐linked silk fibroin hydrogel matrix incorporated with poly(3,4‐ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) are used. The silk fibroin composite is characterized not only by mechanical and electrical conductive properties, but also by its optical transparency in both the visible and ultraviolet regions, and by its biocompatibility with adherent cells. It is demonstrated that the adherent cells, including normal cell‐lined cells and primary neuronal cells, loaded on the nanopallets can be manipulated while faithfully retaining their adhesive properties. By applying voltages via the nanopallets, the voltage‐dependent calcium channels expressed in the cells are selectively stimulated, and this is confirmed by using confocal fluorescent microscopy during manipulation and performing multiangle observations. These features are attributed to both the mobile operation of the transparent nanopallets, and the ability to simultaneously measure electrical signals and perform fluorescent observations.     

Dipole distance for minimum threshold current to stimulate unmyelinated axons with microelectrodes

Excitation thresholds for long nerve or muscle fibers with two point sources parallel to the fiber axis depend on the dipole length. The aim of this study was to find the optimal interelectrode distance for the minimum stimulation current. For a specific electrode-fiber distance (z_el) dipole length is constrained by the energy efficacy of the electrodes requiring small interelectrode distances, and by rather low stimulation currents requiring large dipole distances. Far-field values for optimal dipole distance (approximately 1.4 *z_el) can be explained by the superposition of the positive parts of the activating functions for the monopolar elements of the dipole. A current redistribution effect in a target fiber close to the electrodes shifts the dipole length for threshold stimulation from the theoretical optimal activating function approach value towards greater dipole distances. Spike initiations in straight fibers and retinal ganglion cell axons are investigated.