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Retinal organoid technology enables generation of an inexhaustible supply of three-dimensional retinal tissue from human pluripotent stem cells (hPSCs) for regenerative medicine applications. The high similarity of organoid-derived retinal tissue and transplantable human fetal retina provides an opportunity for evaluating and modeling retinal tissue replacement strategies in relevant animal models in the effort to develop a functional retinal patch to restore vision in patients with profound blindness caused by retinal degeneration. Because of the complexity of this very promising approach requiring specialized stem cell and grafting techniques, the tasks of retinal tissue derivation and transplantation are frequently split between geographically distant teams. Delivery of delicate and perishable neural tissue such as retina to the surgical sites requires a reliable shipping protocol and also controlled temperature conditions with damage-reporting mechanisms in place to prevent transplantation of tissue damaged in transit into expensive animal models. We have developed a robust overnight tissue shipping protocol providing reliable temperature control, live monitoring of the shipment conditions and physical location of the package, and damage reporting at the time of delivery. This allows for shipping of viable (transplantation-competent) hPSC-derived retinal tissue over large distances, thus enabling stem cell and surgical teams from different parts of the country to work together and maximize successful engraftment of organoid-derived retinal tissue. Although this protocol was developed for preclinical in vivo studies in animal models, it is potentially translatable for clinical transplantation in the future and will contribute to developing clinical protocols for restoring vision in patients with retinal degeneration., (© 2019 John Wiley & Sons, Ltd.)

Objective: This overview of ultraviolet (UV) phototoxicity considers the interaction of UVA and short-wavelength VIS light with the retina and retinal pigment epithelium., Methods: The damage mechanisms underlying UV retinal phototoxicity are illustrated with a literature survey and presentation of experimental results., Results: Depending on the wavelength and exposure duration, light interacts with tissue by three general mechanisms: thermal, mechanical, or photochemical. Although the anterior structures of the eye absorb much of the UV component of the optical radiation spectrum, a portion of the UVA band (315-400 nm) penetrates into the retina. Natural sources, such as the sun, emit energetic UV photons in relatively long durations, which typically do not result in energy confinement in the retina, and thus do not produce thermal or mechanical damage but are capable of inducing photochemical damage. Photochemical damage in the retina proceeds through Type 1 (direct reactions involving proton or electron transfers) and Type 2 (reactions involving reactive oxygen species) mechanisms. Commonly used drugs, such as certain antibiotics, nonsteroidal anti-inflammatory drugs, psychotherapeutic agents, and even herbal medicines, may act as photosensitizers that promote retinal UV damage, if they are excited by UVA or visible light and have sufficient retinal penetration., Conclusions: Although the anterior portion of the eye is the most susceptible to UV damage, the retina is at risk to the longer UV wavelengths that propagate through the ocular media. Some phototoxicity may be counteracted or reduced by dietary intake of antioxidants and protective phytonutrients.

This study was conducted to evaluate the effectiveness of a new antifungal drug, micafungin, and standard antifungal drugs against endophthalmitis induced in a rabbit by intravitreal injection of Aspergillus fumigatus, an important fungal pathogen. Effectiveness was evaluated by the preservation of b-wave amplitude at 72 h after injection of the fungus relative to the b-wave amplitude at baseline before any intravitreal injections. A 0.06 ml inoculum of 10(6) conidia of A. fumigatus was injected into the vitreous of the right eye of all rabbits; and, 12 h later, a 0.06 ml solution containing one of 3 antifungal drugs or saline was injected into the vitreous of both eyes. All three antifungal drugs produced significant b-wave preservation at 72 h in infected eyes compared to that in infected eyes receiving saline injections. There was no statistically significant difference between the effects of micafungin and amphotericin B in the right eyes with fungal endophthalmitis, and both produced significantly more preservation of b-wave amplitude than voriconazole. Amphotericin B, but neither micafungin nor voriconazole produced significant reduction of the b-wave amplitude in the left eyes.

Light damage to the retina occurs through three general mechanisms involving thermal, mechanical, or photochemical effects. The particular mechanism activated depends on the wavelength and exposure duration of the injuring light. The transitions between the various light damage mechanism may overlap to some extent. Energy confinement is a key concept in understanding or predicting the type of damage mechanism produced by a given light exposure. As light energy (either from a laser or an incoherent source) is deposited in the retina, its penetration through, and its absorption in, various tissue compartments is determined by its wavelength. Strongly absorbing tissue components will tend to "concentrate" the light energy. The effect of absorbed light energy largely depends on the rate of energy deposition, which is correlated with the exposure duration. If the rate of energy deposition is too low to produce an appreciable temperature increase in the tissue, then any resulting tissue damage necessarily occurs because of chemical (oxidative) reactions induced by absorption of energetic photons (photochemical damage). If the rate of energy deposition is faster than the rate of thermal diffusion (thermal confinement), then the temperature of the exposed tissue rises. If a critical temperature is reached (typically about 10 degrees C above basal), then thermal damage occurs. If the light energy is deposited faster than mechanical relaxation can occur (stress confinement), then a thermoelastic pressure wave is produced, and tissue is disrupted by shear forces or by cavitation-nonlinear effects. Very recent evidence suggests that ultrashort laser pulses can produce tissue damage through nonlinear and photochemical mechanisms; the latter because of two-photon excitation of cellular chromophores. In addition to tissue damage caused directly by light absorption, light toxicity can be produced by the presence of photosensitizing agents. Drugs excited to reactive states by ultraviolet (UV) or visible light produce damage by type I (free radical) and type II (oxygen dependent) mechanisms. Some commonly used drugs, such as certain antibiotics, nonsteroidal anti-inflammatory drugs (NSAIDs), and psychotherapeutic agents, as well as some popular herbal medicines, can produce ocular phototoxicity. Specific cellular effects and damage end points characteristic of light damage mechanisms are described.

Most of the peroxidase activity in the bovine retina is specific to glutathione (GSH) while the choroid contains both GSH peroxidases and ascorbate peroxidase. The GSH peroxidase was clearly separated from ascorbate peroxidase on a cation exchange column. The nonspecific peroxidase activity of hemoproteins accounts for the peroxidase activity detected by ascorbate oxidation. All choroidal hemoproteins contain subunits very similar to those of hemoglobin. The absence of ascorbate peroxidase in the retina indicates that the protective effect of ascorbate against photic injury is not due to its reaction with retinal peroxidase. Therefore removal of H2O2 by ascorbate peroxidase activity in the choroid could be a significant factor in studies where serum ascorbate concentrations are artificially raised far above the normal level concurrent to a very small rise in retinal ascorbate concentration.

Time-dependent effects of laser exposures on rhesus monkey retinal ganglion cells were studied with a Q-switched, doubled Nd:glass laser, which produced 20 nsec pulses of 530-nm light, and a continuous-wave (CW) argon laser (514.5 nm), which produced exposures of 0.1-msec to 0.1-sec duration. Ganglion cell activity was recorded in situ by means of an intraocular electrode. Ganglion cells exposed to a single 20-nsec exposure, at a sublesion intensity, produced a 60-90 sec discharge of action potentials and exhibited a 2 log or greater elevation of light threshold, depending on beam size and intensity. At equivalent energy levels, the longer exposures produced the same or slightly weaker effects. This result is not as straightforward as it seems. Submillisecond flashes bleach no more than 50% of the visual pigment because of photoregeneration. The Dowling-Rushton relation predicts that a 50% bleach should produce only a 1.5 log loss of cone sensitivity. Exposures longer than 1 msec should not photoregenerate pigment (ie, more pigment will be bleached for a given exposure intensity). In view of the probable differences in pigment bleaching, it appears that the Q-switched laser light adapts the cells out of proportion to the visual pigment actually bleached--a single-cell analogue of Rushton's "theta" effect.

Th effects on ganglion cell light responses and spontaneous activity of neurotransmitter candidates, applied by nebulizer spray and iontophoresis, were studied in the isolated carp retina. ACh, GABA, and substance P had strong effects on the ganglion cells; dopamine and the amino acids aspartate, glutamate, and glycine and only weak effects. ACh and substance P exerted their actions even when synaptic transmission was blocked by cobalt chloride, suggesting postsynaptic receptors for those agents on the ganglion cell membrane. The 3 amino acids and dopamine do not appear to act directly on the ganglion cells. The pharmacological sensitivity of ganglion cells was correlated with their physiological response type. About three-quarters of ON/OFF and half of other transiently responding ganglion cells were excited by micromolar concentrations of cholinergic agonists; most ON-center sustained ganglion cells were insensitive. The light response of some of the ACh-sensitive cells could be suppressed by cholinergic antagonists. Substance P generally excited ganglion cells with an ON-component in their light response. GABA inhibited cells of all response types, but affected least the OFF-center tonic cells. In view of these observations, and of corroborating histological evidence, we propose that ACh, GABA, and substance P are neurotransmitters that are released by amacrine cells and affect receptors located on ganglion cells.

Substance P (SP), a neuropeptide, has been found in amacrine cells in a variety of vertebrate retinas. SP excited many of the ganglion cells we sampled in carp retina; many of these ganglion cells are directly excited by cholinergic agonists. It has been reported that SP modulates cholinergic synaptic transmission in some other neuronal systems by inhibiting or desensitizing cholinergic receptors. SP does not act in this way on the ganglion cell cholinergic receptors in isolated carp retina. Pulses of acetylcholine (ACh), applied to sensitive ganglion cells by microiontophoresis, were set to elicit consistent cellular responses. Application of SP to the retina through a nebulizing system (final concentration about 10(-6) M) did not reduce the excitation produced by the ACh pulse. In earlier work, we showed that the cholinergic receptors were nicotinic; in the present study, SP occasionally excited cells after the retina was treated with a nicotinic cholinergic antagonist, gallamine triethiodide. SP appears to excite cholinergic-sensitive ganglion cells at a postsynaptic site other than their nicotinic cholinergic receptor and via a pathway that does not require a presynaptic, SP-sensitive, ACh-releasing interneuron.