The immunology of corneal limbal stem cells: the up-to-date approach to stem cell transplantation.

Limbal epithelial stem cells (LSC, LESC) are multipotent cells used as regenerative treatment of the cornea in patients with limbal epithelial stem cell deficiency (LSCD, LESCD). There are different types of stem cell grafting including cultivated limbal epithelial transplantation (CET) and simple limbal epithelial transplantation (SLET). The outcomes of the techniques have been assessed as similar, with differences in the sample size required during the procedures. The most important culture components for stem cell cultivation include 3T3 murine fibroblasts, human amniotic membrane (HAM), fibrin gel, and culture medium. The culture medium may be enriched with serum or not; however, xenobiotic-free materials are preferred because of the low risk of pathogen transmission. Multiple studies have defined molecules important for maintaining the function of LSC including C/EBP δ, Bmi-1, p63 α, interleukins (IL-6), epithelial structural proteins - keratins, and antibodies against epidermal growth factor receptor (EGFR). The cell phenotype of LSC has been described with factors of transplantation success rate such as a high percentage of p63 positive cells. The article emphasizes the role of recipient tissue preparation, modern cultivation techniques and pathophysiological processes in LSC transplantation effectiveness.

Novel therapies for diabetic retinopathy.

Diabetes mellitus (DM) is a metabolic disease characterized by high blood glucose levels as well as microvascular and macrovascular changes. According to the latest statistics the growth of DM incidence is very fast. Diabetic retinopathy (DR) - one of the common DM complications - is the leading cause of blindness among professionally active people. Traditional treatment of DR including drugs controlling hyperglycemia, laser therapy, vitrectomy, and intravitreal injections of anti-VEGF is effectively administered with the effect of neovascularization and macular edema prevention. However, new potential DR therapies - focusing on a longer therapeutic effect and potentially fewer side effects - are being widely investigated. Gene therapy - targeting retinal vasculopathy or targeting retinal protection, mesenchymal stem cell injections, SGLT2 inhibitors, and islet cell transplantation have been proved to stop DR progression. The majority of the new treatment research was performed on an animal model and did not reach the final study stage. A further future human model and randomized studies with optimized delivery vectors will hopefully confirm positive outcomes of the new DR therapies.

Novel approach to antiangiogenic factors in age-related macular degeneration therapy.

Age-related macular degeneration (AMD) is a leading cause of irreversible vision loss among the population above 85 worldwide. There are two main types of AMD: neovascular and dry AMD. Neovascular AMD leads to macular changes resulting from abnormal choroidal neovascularization. Untreated neovascular AMD leads to scar formation and irreversible sight deterioration. Dry AMD in consequence leads to atrophic changes of the macula. The last decades brought a breakthrough in the therapy of neovascular age-related macular degeneration by introduction of, firstly, photodynamic therapy and, later, anti-VEGF agents administered intravitreally in order to stop neoangiogenesis. However, the treatment of dry AMD is still challenging. Among the directions in dry AMD treatment, the most promising are complement cascade inhibitors and complement cascade targeted gene therapy. In the article we outline the main directions in up-to-date experimental and practical approaches to wet and dry AMD therapy with the emphasis on antiangiogenic factors and gene therapy focused on the inhibition of pathological angiogenesis.

New perspectives of immunomodulation and neuroprotection in glaucoma.

Glaucoma is the neurodegenerative disease of retinal ganglion cells. The main risk factor for glaucoma is increased intraocular pressure. The processes leading to cell death due to presence of the injury factor comprise multiple molecular mechanisms, as well as the immunological response. The knowledge of immunological mechanisms occurring in glaucomatous degeneration makes it possible to introduce glaucoma treatment modulating the cellular degradation. The glaucoma treatment of the future will make it possible not only to lower the intraocular pressure, but also to moderate the intracellular mechanisms in order to prevent retinal cell degeneration. Citicoline is a drug modulating glutamate excitotoxicity that is already in use. Rho kinase inhibitors were found to stimulate neurite growth and axon regeneration apart from lowering intraocular pressure. The complementary action of brimonidine is to increase neurotrophic factor (NTF) concentrations and inhibit glutamate toxicity. Immunomodulatory therapies with antibodies and gene therapies show promising effects in the current studies. The supplementation of NTFs prevents glaucomatous damage. Resveratrol and other antioxidants inhibit reactive oxygen species formation. Cell transplantation of stem cells, Schwann cells and nerve extracts was reported to be successful so far. Our review presents the most promising new strategies of neuroprotection and immunomodulation in glaucoma.

Retinal Photoreceptors and Microvascular Changes in the Assessment of Diabetic Retinopathy Progression: A Two-Year Follow-Up Study.

BACKGROUND: With the increasing global incidence of diabetes mellitus (DM), diabetic retinopathy (DR) has become one of the leading causes of blindness in developed countries. DR leads to changes in retinal neurons and microcirculation. Rtx1TM (Imagine Eyes, Orsay, France) is a retinal camera that allows histological visualisations of cones and retinal microcirculation throughout the DM duration. OBJECTIVE: This study aimed to analyse the cones and retinal microvascular changes in 50 diabetic individuals and 18 healthy volunteers. The patients participated in the initial visit and two follow-up appointments, one and two years after the study, beginning with Rtx1TM image acquisition, visual acuity assessment, macular OCT scans and blood measurements. RESULTS: The study revealed significant differences in the cone density, mosaic arrangement and vascular morphology between healthy and diabetic patients. The final measurements showed decreased photoreceptor and microvascular parameters in the DR group compared with the control group. Furthermore, in the 2-year follow-up, both groups' Rtx1TM-acquired morphological changes were statistically significant. CONCLUSIONS: Rtx1TM technology was successfully used as a non-invasive method of photoreceptors and retinal vasculature assessment over time in patients with diabetic retinopathy. The study revealed a trend toward more vascular morphological changes occurring over time in diabetic patients.

The Relation between Body Mass Index and Retinal Photoreceptor Morphology and Microvascular Changes Measured with Adaptive Optics (rtx1) High-Resolution Imaging.

BACKGROUND: Overweight and obese patients are at risk for diabetes, cardiovascular disorders, and microvascular complications. The rtx1TM (Imagine Eyes, France) is a microscope that allows near histological visualizations of cones and retinal microcirculation. OBJECTIVE: This study analysed the cones and retinal microvascular changes in a group of 47 healthy women with different BMI values. Participants were divided into 2 groups: the BMI group (28 women with BMI >/25) and the control group (19 lean women with BMI <25). RESULTS: The lumen and diameter of retinal arteries were not significantly different between groups. There were significant differences in the thickness of arteriole walls. The WLR and WCSA values differed significantly between the control and BMI groups (for WLR 0.25 ± 0.03 vs. 0.29 ± 0.03, p < 0.001; for WCSA 4136.7 ± 1140.0 vs. 5217.3 ± 944.0, respectively, p < 0.001). In healthy eyes, cone density and morphology were not affected by weight. CONCLUSIONS: Retinal image analysis with rtx1 offers a novel noninvasive measurement of early changes in retinal vasculature that are not detectable during routine clinical examination. Abnormalities of retinal arterioles found by rtx1™examination should be considered as a strong risk factor for cardiovascular changes resulting from overweight and obesity.