Neovascular age-related macular degeneration (nvAMD) is characterized by choroidal neovascularization (CNV), which leads to retinal pigment epithelial (RPE) cell and photoreceptor degeneration and blindness if untreated. Since blood vessel growth is mediated by endothelial cell growth factors, including vascular endothelial growth factor (VEGF), treatment consists of repeated, often monthly, intravitreal injections of anti-angiogenic biopharmaceuticals. Frequent injections are costly and present logistic difficulties; therefore, our laboratories are developing a cell-based gene therapy based on autologous RPE cells transfected ex vivo with the pigment epithelium derived factor (PEDF), which is the most potent natural antagonist of VEGF. Gene delivery and long-term expression of the transgene are enabled by the use of the non-viral Sleeping Beauty (SB100X) transposon system that is introduced into the cells by electroporation. The transposase may have a cytotoxic effect and a low risk of remobilization of the transposon if supplied in the form of DNA. Here, we investigated the use of the SB100X transposase delivered as mRNA and showed that ARPE-19 cells as well as primary human RPE cells were successfully transfected with the Venus or the PEDF gene, followed by stable transgene expression. In human RPE cells, secretion of recombinant PEDF could be detected in cell culture up to one year. Non-viral ex vivo transfection using SB100X-mRNA in combination with electroporation increases the biosafety of our gene therapeutic approach to treat nvAMD while ensuring high transfection efficiency and long-term transgene expression in RPE cells.

Oxidative stress (OS) is involved in the pathogenesis of retinal neurodegenerative diseases such as age-related macular degeneration (AMD) and diabetic retinopathy (DR) and an important target of therapeutic treatments. New therapeutics are tested in vivo despite limits in terms of transferability and ethical concerns. Retina cultures using human tissue can deliver critical information and significantly reduce the number of animal experiments along with increased transferability. We cultured up to 32 retina samples derived from one eye, analyzed the model’s quality, induced OS, and tested the efficiency of antioxidative therapeutics. Bovine, porcine, rat, and human retinae were cultured in different experimental settings for 3–14 d. OS was induced by a high amount of glucose or hydrogen peroxide (H₂O₂) and treated with scutellarin, pigment epithelium-derived factor (PEDF), and/or granulocyte macrophage colony-stimulating factor (GM-CSF). The tissue morphology, cell viability, inflammation, and glutathione level were determined. The retina samples showed only moderate necrosis (23.83 ± 5.05 increased to 27.00 ± 1.66 AU PI-staining over 14 d) after 14 days in culture. OS was successfully induced (reduced ATP content of 288.3 ± 59.9 vs. 435.7 ± 166.8 nM ATP in the controls) and the antioxidants reduced OS-induced apoptosis (from 124.20 ± 51.09 to 60.80 ± 319.66 cells/image after the scutellarin treatment). Enhanced mammalian animal and human retina cultures enable reliable, highly transferable research on OS-triggered age-related diseases and pre-clinical testing during drug development.

Among ocular infections, trachoma is the main cause of blindness. Repeated conjunctival Chlamydia trachomatis infections lead to trichiasis, corneal opacification, and visual impairment. Surgery is often needed to relieve discomfort and preserve vision; however, a high postoperative trachomatous trichiasis (PTT) rate has been observed in various settings. We wanted to know why, whether PTT rates could be reduced, and how to manage the PTT that occurs. We performed a search of the literature. Of 217 papers screened, 59 studies were identified for inclusion as potentially relevant, the majority having been excluded for not directly concerning PTT in humans. Preventing PTT is a major challenge. Only one published trial, the STAR trial in Ethiopia, has reported a cumulative PTT rate <10% one year after surgery. The literature on the management of PTT is sparse. Though no PTT management guidelines are available, high-quality surgery with a low rate of unfavorable outcomes for PTT patients is likely to require enhanced training of a smaller group of highly-skilled surgeons. Based on the surgical complexity and the authors' own experience, the pathway for patients suffering from PTT should be studied further for improvement.

Background/Objectives: The goal of this study was to show the consistency between the inflammatory and anti-inflammatory response in the early postoperative period after corneal refractive surgery (LASIK and PRK), represented by the levels of specific cytokines in the tear film. Materials/Methods: A total of 68 myopic eyes (31 in the LASIK and 37 in the PRK group) had 3 samples of tear film taken, one before surgery (t₀), another 1 h after surgery (t₁) and the third 24 h after surgery (t₂). The tear samples were then analyzed by flow cytometry in order to determine the levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, IFN-γ, TNF-α) and the levels of anti-inflammatory cytokines (IL-4, IL-10). Results: A statistically significant positive correlation was found between the concentrations of all pro-inflammatory cytokines (IL-1β, IL-6, IL-8, IFN-γ, TNF-α) and all anti-inflammatory cytokines (IL-4, IL-10), respectively, for both t₁ and t₂ in the group of eyes that underwent LASIK. In the PRK group, there was a statistically significant positive correlation for all cytokines (pro-inflammatory and anti-inflammatory) at t₁; however at t_2, there was no correlation between IL-1β and IL-10, IL-6 and IL-10, IL-8 and IL-10, and IFN-γ and IL-10. In contrast, for other correlations between pro-inflammatory and anti-inflammatory cytokines in the same observation time (t₂), a positive correlation was observed. Conclusions: There is a consistency between the inflammatory and anti-inflammatory responses in the early postoperative period after refractive surgery, which persists longer after LASIK compared to PRK. This difference may explain the different clinical courses in patients after undergoing these procedures.