Curcumin as a Perspective Protection for Retinal Pigment Epithelium during Autophagy Inhibition in the Course of Retinal Degeneration.

Defective autophagy in the retinal pigment epithelium (RPE) is involved in retinal degeneration, mostly in the course of age-related macular degeneration (AMD), which is an increasingly prevalent retinal disorder, eventually leading to blindness. However, most autophagy activators own serious adverse effects when administered systemically. Curcumin is a phytochemical, which induces autophagy with a wide dose-response curve, which brings minimal side effects. Recent studies indicating defective autophagy in AMD were analyzed. Accordingly, in this perspective, we discuss and provide some evidence about the protective effects of curcumin in preventing RPE cell damage induced by the autophagy inhibitor 3-methyladenine (3-MA). Cells from human RPE were administered the autophagy inhibitor 3-MA. The cell damage induced by 3-MA was assessed at light microscopy by hematoxylin & eosin, Fluoro Jade-B, and ZO1 immunohistochemistry along with electron microscopy. The autophagy inhibitor 3-MA produces cell loss and cell degeneration of RPE cells. These effects are counteracted dose-dependently by curcumin. In line with the hypothesis that the autophagy machinery is key in sustaining the integrity of the RPE, here we provide evidence that the powerful autophagy inhibitor 3-MA produces dose-dependently cell loss and cell degeneration in cultured RPE cells, while inhibiting autophagy as shown by LC3-II/LC3-I ratio and gold-standard assessment of autophagy through LC3-positive autophagy vacuoles. These effects are prevented dose-dependently by curcumin, which activates autophagy. These data shed the perspective of validating the role of phytochemicals as safe autophagy activators to treat AMD.

Autophagy Activation Promoted by Pulses of Light and Phytochemicals Counteracting Oxidative Stress during Age-Related Macular Degeneration.

The seminal role of autophagy during age-related macular degeneration (AMD) lies in the clearance of a number of reactive oxidative species that generate dysfunctional mitochondria. In fact, reactive oxygen species (ROS) in the retina generate misfolded proteins, alter lipids and sugars composition, disrupt DNA integrity, damage cell organelles and produce retinal inclusions while causing AMD. This explains why autophagy in the retinal pigment epithelium (RPE), mostly at the macular level, is essential in AMD and even in baseline conditions to provide a powerful and fast replacement of oxidized molecules and ROS-damaged mitochondria. When autophagy is impaired within RPE, the deleterious effects of ROS, which are produced in excess also during baseline conditions, are no longer counteracted, and retinal degeneration may occur. Within RPE, autophagy can be induced by various stimuli, such as light and naturally occurring phytochemicals. Light and phytochemicals, in turn, may synergize to enhance autophagy. This may explain the beneficial effects of light pulses combined with phytochemicals both in improving retinal structure and visual acuity. The ability of light to activate some phytochemicals may further extend such a synergism during retinal degeneration. In this way, photosensitive natural compounds may produce light-dependent beneficial antioxidant effects in AMD.

The Essential Role of Light-Induced Autophagy in the Inner Choroid/Outer Retinal Neurovascular Unit in Baseline Conditions and Degeneration.

The present article discusses the role of light in altering autophagy, both within the outer retina (retinal pigment epithelium, RPE, and the outer segment of photoreceptors) and the inner choroid (Bruch's membrane, BM, endothelial cells and the pericytes of choriocapillaris, CC). Here autophagy is needed to maintain the high metabolic requirements and to provide the specific physiological activity sub-serving the process of vision. Activation or inhibition of autophagy within RPE strongly depends on light exposure and it is concomitant with activation or inhibition of the outer segment of the photoreceptors. This also recruits CC, which provides blood flow and metabolic substrates. Thus, the inner choroid and outer retina are mutually dependent and their activity is orchestrated by light exposure in order to cope with metabolic demand. This is tuned by the autophagy status, which works as a sort of pivot in the cross-talk within the inner choroid/outer retina neurovascular unit. In degenerative conditions, and mostly during age-related macular degeneration (AMD), autophagy dysfunction occurs in this area to induce cell loss and extracellular aggregates. Therefore, a detailed analysis of the autophagy status encompassing CC, RPE and interposed BM is key to understanding the fine anatomy and altered biochemistry which underlie the onset and progression of AMD.

A Re-Appraisal of Pathogenic Mechanisms Bridging Wet and Dry Age-Related Macular Degeneration Leads to Reconsider a Role for Phytochemicals.

Which pathogenic mechanisms underlie age-related macular degeneration (AMD)? Are they different for dry and wet variants, or do they stem from common metabolic alterations? Where shall we look for altered metabolism? Is it the inner choroid, or is it rather the choroid-retinal border? Again, since cell-clearing pathways are crucial to degrade altered proteins, which metabolic system is likely to be the most implicated, and in which cell type? Here we describe the unique clearing activity of the retinal pigment epithelium (RPE) and the relevant role of its autophagy machinery in removing altered debris, thus centering the RPE in the pathogenesis of AMD. The cell-clearing systems within the RPE may act as a kernel to regulate the redox homeostasis and the traffic of multiple proteins and organelles toward either the choroid border or the outer segments of photoreceptors. This is expected to cope with the polarity of various domains within RPE cells, with each one owning a specific metabolic activity. A defective clearance machinery may trigger unconventional solutions to avoid intracellular substrates' accumulation through unconventional secretions. These components may be deposited between the RPE and Bruch's membrane, thus generating the drusen, which remains the classic hallmark of AMD. These deposits may rather represent a witness of an abnormal RPE metabolism than a real pathogenic component. The empowerment of cell clearance, antioxidant, anti-inflammatory, and anti-angiogenic activity of the RPE by specific phytochemicals is here discussed.

Riboflavin osmolar modification for transepithelial corneal cross-linking.

PURPOSE: To investigate the influence of osmolarity on the transepithelial permeability of riboflavin solutions in a cross-linking procedure. METHODS: Several riboflavin 0.1% solutions that contained different NaCl and benzalkonium chloride (BAC) concentrations were applied to 36 rabbit eyes for 30 min. To serve as a control, the epithelium was removed in group A (standard protocol). The groups then received the following solutions: (A) riboflavin 0.1% in NaCl 0.9% solution; (B) riboflavin 0.1% in NaCl 0.44% solution with BAC 0.02%; (C) riboflavin 0.1% in NaCl 0.44% solution with BAC 0.01%; (D) riboflavin 0.1% in NaCl 0.44% solution without BAC; (E) riboflavin 0.1% in NaCl 0.9% solution with BAC 0.02%; and (F) riboflavin 0.1% in NaCl 0.9% solution without BAC. Six eyes in each group were treated. The absorption coefficients of the corneas were measured to characterize the riboflavin penetration into the cornea. RESULTS: There is a large difference in the transepithelial riboflavin penetration of riboflavin 0.1% + BAC 0.02% solutions that contain different NaCl concentrations (NaCl 0.9% versus NaCl 0.44%). The absorption coefficients differed by more than a factor of two (P = 0.004). No statistically significant difference was found between riboflavin 0.1% in NaCl 0.44% solution containing BAC 0.02% and BAC 0.01%. Compared to the standard protocol, these solutions resulted in an absorption coefficient of 37% (BAC 0.02%) and 33% (BAC 0.01%) of the standard epithelium-off procedure. CONCLUSION: The transepithelial riboflavin solution should contain no dextran, but it should include 0.01% BAC and 0.44% NaCl to promote the permeability of riboflavin through the epithelium, resulting in a sufficient concentration of riboflavin in the corneal stroma.

Correction of presbyopia in hyperopia with a center-distance, paracentral-near technique using the Technolas 217z platform.

PURPOSE: To analyze the results of hyperopic patients treated with a peripheral presbyLASIK algorithm for the correction of presbyopia. METHODS: The study included 44 eyes of 22 hyperopic patients treated with a peripheral presbyLASIK technique using a Technolas 217z excimer laser. Mean patient age was 56 years (range: 47 to 72 years), mean preoperative spherical equivalent refraction was +1.21 +/- 0.77 diopters (D) (range: +0.50 to +4.00 D), and mean spectacle near addition was +1.76 +/- 0.42 D (range: +1.00 to +2.75 D). The Peripheral Multifocal LASIK (PML) ablation pattern creates a multifocal corneal profile over a 6.5-mm diameter, performing the distance correction first in a 6-mm optical zone and then near correction in a 6.5-mm zone. Main outcome measures were uncorrected visual acuity (UCVA) and best spectacle-corrected visual acuity (BSCVA) for near and distance, spherical equivalent refraction, contrast sensitivity, and corneal aberrations. RESULTS: Six months postoperatively, mean binocular UCVA was 1.06 +/- 0.13 for distance and 0.84 +/- 0.14 for near. Mean postoperative spherical equivalent refraction was -0.42 D (range: -1.12 to +0.87 D). Two (4.5%) eyes lost 1 line of BSCVA for distance and near vision, and 20 (45%) eyes gained 1 line of distance BSCVA. Contrast sensitivity decreased for 3, 6, 12, and 18 cycles/degree. Corneal aberration analysis showed a slight increase in coma and decrease in spherical aberration. CONCLUSIONS: The peripheral presbyLASIK technique used in this study is a safe and efficient treatment that may improve functional near vision in presbyopic patients with low and moderate hyperopia (from +0.50 to +3.00 D).

Sequential ablation approach to the correction of mixed astigmatism.

PURPOSE: To evaluate the safety, efficacy, and stability of LASIK, using positive cylinder and negative sphere nomograms in sequence (sequential ablation) to correct mixed astigmatism. METHODS: This prospective study included 40 eyes of 20 patients with mixed astigmatism. Patients underwent bilateral sequential ablation LASIK using the Technolas 217 excimer laser (Bausch & Lomb Surgical, Rochester, NY). The main outcome measures, uncorrected visual acuity (UCVA) and best spectacle-corrected visual acuity (BSCVA), were evaluated 3 and 12 months after surgery. RESULTS: Preoperative astigmatism ranged from +1.75 to +6.00 diopters (D) and negative sphere from -0.50 to -3.00 D. The postoperative refraction at 3 months remained unchanged at 1 year postoperative in all patients. A total of 32 (80%) eyes showed no significant residual astigmatism (<0.50 D); the remaining 8 (20%) eyes had 0.50 to 1.00 D of residual astigmatism. Residual negative sphere was present in 2 eyes of 2 patients with a planned monovision target. In the remaining 38 (95%) eyes, no significant residual negative sphere was present. Sixteen (40%) eyes had one line of improvement in BSCVA. No eye lost lines of visual acuity. The efficacy index shows that uncorrected vision after surgery is equal or better than corrected vision before surgery. Less corneal tissue is removed and fewer laser spots are required compared to other techniques for the correction of mixed astigmatism. CONCLUSIONS: The sequential ablation approach to the correction of mixed astigmatism was efficacious, safe, and stable 1 year after surgery.