Mapping of bionic array electric field focusing in plasmid DNA-based gene electrotransfer.

Molecular medicine through gene therapy is challenged to achieve targeted action. This is now possible utilizing bionic electrode arrays for focal delivery of naked (plasmid) DNA via gene electrotransfer. Here, we establish the properties of array-based electroporation affecting targeted gene delivery. An array with eight 300 µm platinum ring electrodes configured as a cochlear implant bionic interface was used to transduce HEK293 cell monolayers with a plasmid-DNA green fluorescent protein (GFP) reporter gene construct. Electroporation parameters were pulse intensity, number, duration, separation and electrode configuration. The latter determined the shape of the electric fields, which were mapped using a voltage probe. Electrode array-based electroporation was found to require ~100 × lower applied voltages for cell transduction than conventional electroporation. This was found to be due to compression of the field lines orthogonal to the array. A circular area of GFP-positive cells was created when the electrodes were ganged together as four adjacent anodes and four cathodes, whereas alternating electrode polarity created a linear area of GFP-positive cells. The refinement of gene delivery parameters was validated in vivo in the guinea pig cochlea. These findings have significant clinical ramifications, where spatiotemporal control of gene expression can be predicted by manipulation of the electric field via current steering at a cellular level.

A method for the generation of complex vibrotactile stimuli.

A method is described utilizing computer-generated sine wave data and purpose-built hardware to generate a complex vibrotactile stimulus. Two sine waves of different frequency were summed to produce a complex waveform with two temporal components, a high frequency component and a low frequency beat component. The computer-generated data points for each of the two component sine waves were downloaded to two banks of static memory in a dual synchronous arbitrary function generator. The data points in memory were fed to two 12-bit digital-to-analogue converters which sent the two analogue sine wave signals to a summing amplifier where the two sine waves were added. This method provides a complex waveform that can be gated on and off, has a fixed frequency ratio of the component sine waves and no phase drift between the component waves. Addition of the separate sine waves in a summing amplifier allows for easy alteration of the amplitude ratio of the sine waves. The output of the summing amplifier is sent to a feedback controlled mechanical stimulator, thereby allowing the stimulus to be presented to the skin of human subjects and experimental animals.