Towards a Retina Prosthesis Model: Neurons on Microphotodiode Arrays In Vitro

Abstract

Various neurodegenerative diseases of the retina lead to blindness. Since no pharmacological or gene therapy is available, the alternative concept of neuroprostheses has stimulated the development of micromachined photovoltaic devices. One strategy aims to replace degenerated photoreceptors by microphotodiode arrays (MPDA). MPDAs in the subretinal space shall transform light stimuli into electrical current for the stimulation of still unhampered retinal neurons. MPDAs were fabricated as silicon based multilayered PIN microstructures comprising titanium nitride stimulation electrodes of an area of 8 × 8 μm2. In an effort to develop meaningful tools to evaluate functional MPDA biocompatibility, a novel in vitro system was designed. Retinal cells were cultured on MPDAs, while current generation in MPDAs as in the eye was imitated by light emitting diodes (LED) operating at different wavelengths. For analysis epifluorescence and scannning electron microscopy was employed. MPDAs displayed increasing current delivery with increasing illumination. Surface modification of MPDAs including oxygen plasma treatment and adsorption of polyanions together with laminin were found to render the MPDA surface permissive for cell adhesion. Cell vitality tests using fluorescence markers revealed no adverse effects of optoelectric stimulation via LED/MPDAs. In addition, neurite formation and the expression of differentiation antigen 2A10 were unaffected after stimulation. Optoelectric stimulation allowed regular differentiation of various retinal cell types. In summary, the data provide the first evidence that optoelectric stimulation via MPDAs does not hamper cellular integrity under the experimental conditions chosen. The results support the concept of microphotodiodes as a retinal prosthesis.