Document Type masters report Author Name Huang, Fei URN etd-082412-105249 Title Electrophysiological Studies of a Retinal Prosthetic Prototype Degree MS Department Chemistry & Biochemistry Advisors Christopher Lambert, Advisor Keywords Self Assembled Monolayer Electrophysiology Date of Presentation/Defense 2012-09-26 Availability restricted
Retinal prostheses are becoming a viable therapy for inner retinal degeneration caused by age related macular degeneration (AMD) and retinitis pigmentosa (RP). The majority of ocular and periocular prosthetic devices employ photodiodes and a microelectrode interface to convert light into a nerve impulse within the retina. Problems with this design include the need of an external power source, the lack of biocompatibility of the microelectrode array, and the need for complicated surgical procedures. Self-assembled monolayer (SAM) technology offers an alternative strategy, where neurons can be stimulated by light activation of a single layer of a photovoltaic SAM. We have developed a SAM structure where the photoexcitable dye 2-[2-[4-(dibutylamino)phenyl]ethenyl]-3-caboxymethylbenzothiazolium bromide (NK5962) was covalently immobilized to an indium tin oxide (ITO) and 3-(aminopropyl) trimethoxysilane (APTMS) surface. The NK562 derivatized surface was characterized through contact angle goniometry, electrochemical impedance spectroscopy, grazing angle infrared spectroscopy, and ultraviolet-visible absorption spectrophotometry. NG108-15 neurons were differentiated onto the surface and neural responses from electrical stimulation and photostimulation of the system were measured using whole-cell current and voltage clamp methodologies. We found an average 2.9 mV decrease in NG108-15 threshold potential for every 10 mV increase in ITO surface potential. Following photostimulation, there was a 1.8±0.2 fold increase (p < 0.05) in the sodium channel current amplitude and a 2.00 ± 0.22 fold increase (p < 0.05 ) in voltage amplitude of NG108-15 neurons on the ITO-APTMS-NK5962 surface due to transfer of energy from the excited dye surface to the attached neurons. The degree of photostimulation decreased upon using 344, 430, and 603 nm optical filters to block increasing amounts of the wavelengths of incident light capable of being absorbed by NK5962. The sodium current amplitude slightly increased at 50% transmittance of incident light relative to 100% transmittance, then sharply decreased at 12.5%, 6.25%, 3.13% transmittance. Upon addition of tetrodotoxin (TTX), sodium channel blockage was observed and portrayed by decreased sodium current and voltage response amplitudes, validating the voltage and current clamp results described above. Our findings indicate that the NK5962 photoelectric film shows promise as an implant for restoring light sensitivity to the retina.
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