Photovoltaic, wireless wide-field epiretinal prosthesis to treat retinitis pigmentosa.

PURPOSE: To develop and evaluate a photovoltaic, wireless wide-field epiretinal prosthesis for the treatment of retinitis pigmentosa. METHODS: A mosaic array of thinned silicon-based photodiodes with integrated thin-film stimulation electrodes was fabricated with a flexible polyimide substrate film to form a film-based miniaturized electronic system with wireless optical power and signal transmission and integrated electrostimulation. Manufactured implants were characterized with respect to their optoelectronic performance and biocompatibility following DIN EN ISO 10993. RESULTS: A 14 mm diameter prosthesis containing 1276 pixels with a maximum sensitivity at a near infrared wavelength of 905 nm and maximized stimulation current density 30-50 µm below the electrodes was developed for direct activation of retinal ganglion cells during epiretinal stimulation. Fabricated prostheses demonstrated mucosal tolerance and the preservation of both metabolic activity, proliferation and membrane integrity of human fibroblasts as well as the retinal functions of bovine retinas. Illumination of the prosthesis, which was placed epiretinally on an isolated perfused bovine retina, with infrared light resulted in electrophysiological recordings reminiscent of an a-wave (hyperpolarization) and b-wave (depolarization). CONCLUSIONS: A photovoltaic, wireless wide-field epiretinal prosthesis for the treatment of retinitis pigmentosa using near infrared light for signal transmission was designed, manufactured and its biocompatibility and functionality demonstrated in vitro and ex vivo.

A HET-CAM based vascularized intestine tumor model as a screening platform for nano-formulated photosensitizers.

The development of new tumor models for anticancer drug screening is a challenge for preclinical research. Conventional cell-based in vitro models such as 2D monolayer cell cultures or 3D spheroids allow an initial assessment of the efficacy of drugs but they have a limited prediction to the in vivo effectiveness. In contrast, in vivo animal models capture the complexity of systemic distribution, accumulation, and degradation of drugs, but visualization of the individual steps is challenging and extracting quantitative data is usually very difficult. Furthermore, there are a variety of ethical concerns related to animal tests. In accordance with the 3Rs principles of Replacement, Reduction and Refinement, alternative test systems should therefore be developed and applied in preclinical research. The Hen's egg test on chorioallantoic membrane (HET-CAM) model provides the generation of vascularized tumor spheroids and therefore, is an ideal test platform which can be used as an intermediate step between in vitro analysis and preclinical evaluation in vivo. We developed a HET-CAM based intestine tumor model to investigate the accumulation and efficacy of nano-formulated photosensitizers. Irradiation is necessary to activate the phototoxic effect. Due to the good accessibility of the vascularized tumor on the CAM, we have developed a laser irradiation setup to simulate an in vivo endoscopic irradiation. The study presents quantitative as well as qualitative data on the accumulation and efficacy of the nano-formulated photosensitizers in a vascularized intestine tumor model.

Diffraction-based technology for the monitoring of contraction dynamics in 3D and 2D tissue models.

We present a novel optical device developed for the monitoring of dynamic behavior in extended 3D-tissue models in various culture environments based on variations in their speckle patterns. The results presented point out the benefit of the technology in terms of detection, accuracy, sensitivity and a reasonable read-out speed as well as reproducibility for the measurements and monitoring of cardiac contractions. We show that the optical read-out technology is suitable for long time monitoring and for drug screening. The method is discussed and compared to other techniques, in particular calcium imaging. The device is flexible and easily adaptable to 2D and 3D-tissue model screenings using different culture environments. The technology can be parallelized for automated read-out of different multi-well-plate formats.

Sub-cellular tumor identification and markerless differentiation in the rat brain in vivo by multiphoton microscopy.

OBJECTIVE/BACKGROUND: Aim of the current study was to localize and differentiate between tumor (glioma) and healthy tissue in rat brains on a cellular level. Near-infrared multiphoton microscopy takes advantage of the simultaneous absorption of two or more photons to analyze various materials such as cell and tissue components via the observation of endogenous fluorophores such as NAD(P)H, FAD, porphyrins, melanin, elastin, and collagen, with a very high resolution, without inducing the problems of photo-bleaching on out-of-focus areas. METHODS: In vitro and in vivo studies on healthy rat brains as well as C6 glioma cell line allografts have been performed. Near-infrared laser pulses (λ = 690-1060 nm, τ ~140 fs) generated by an ultrafast Ti:Sapphire tunable laser system (Chameleon, Coherent GmbH, Santa Clara, CA) were coupled into a laser scanning microscope (LSM 510 META, Carl Zeiss, Germany) to observe high quality images. RESULTS: Several image acquisitions have been performed by varying the zoom scale of the multiphoton microscope, image acquisition time and the wavelength (765, 840 nm) to detect various tissue components. With a penetration depth of ~200 µm in vitro and about 30-60 µm in vivo into the brain tissue it was possible to differentiate between tumor and healthy brain tissue even through thin layers of blood. CONCLUSION: Near-infrared multiphoton microscopy allows the observation and possibly differentiation between tumor (glioma) and healthy tissue in rat brains on a cellular level. Our findings suggest that a further miniaturization of this technology might be very useful for scientific and clinical applications in neurosurgery.

Ultraviolet femtosecond laser creation of corneal flap.

PURPOSE: To study parameters for ocular femtosecond laser surgery in terms of process efficiency and safety aspects using ultraviolet (UV) femtosecond laser pulses. METHODS: Studies on corneal surgery and flap processing on enucleated porcine eyes were performed using a newly developed ytterbium-doped gain media laser source. Ultraviolet femtosecond laser pulses centered at a wavelength of 345 nm and working at a repetition rate of 100 kHz were generated by the third harmonics of the 1035-nm fundamental wavelength. RESULTS: Flaps with a diameter of 6 mm and a thickness of 100 microm were created in less than 2 minutes with low energy pulses. Transmissions and spectral measurements were performed during flap processing. Less than 2% UV radiation reaches the retina during corneal flap processing. A detectable transmittance towards the retina of visible light centered on 440 nm was found for UV pulses. CONCLUSIONS: Ultraviolet corneal refractive surgery is a novel procedure and has the potential to be an alternative to infrared refractive surgery considering safety aspects.

Sub-100 nm nanostructuring of silicon by ultrashort laser pulses.

Techniques based on laser scanning microscopes for nanoprocessing of periodic structures on silicon with ultra-short laser pulses have been developed. Ripples of 800-900 nm spacing were obtained after laser irradiation at a wavelength of 1040 nm, a repetition rate of 10 kHz and a fluence of 2 J/cm2 in air. Smaller features of 70-100nm spacing were achieved in oil at a wavelength of 800 nm, a repetition rate of 90 MHz and a fluence of 200-300 mJ/cm2 by using a high numerical focusing objective.