Effects of Rho-Associated Kinase (Rock) Inhibitors (Alternative to Y-27632) on Primary Human Corneal Endothelial Cells.

(1) Rho-associated coiled-coil protein kinase (ROCK) signaling cascade impacts a wide array of cellular events. For cellular therapeutics, scalable expansion of primary human corneal endothelial cells (CECs) is crucial, and the inhibition of ROCK signaling using a well characterized ROCK inhibitor (ROCKi) Y-27632 had been shown to enhance overall endothelial cell yield. (2) In this study, we compared several classes of ROCK inhibitors to both ROCK-I and ROCK-II, using in silico binding simulation. We then evaluated nine ROCK inhibitors for their effects on primary CECs, before narrowing it down to the two most efficacious compounds-AR-13324 (Netarsudil) and its active metabolite, AR-13503-and assessed their impact on cellular proliferation in vitro. Finally, we evaluated the use of AR-13324 on the regenerative capacity of donor cornea with an ex vivo corneal wound closure model. Donor-matched control groups supplemented with Y-27632 were used for comparative analyses. (3) Our in silico simulation revealed that most of the compounds had stronger binding strength than Y-27632. Most of the nine ROCK inhibitors assessed worked within the concentrations of between 100 nM to 30 µM, with comparable adherence to that of Y-27632. Of note, both AR-13324 and AR-13503 showed better cellular adherence when compared to Y-27632. Similarly, the proliferation rates of CECs exposed to AR-13324 were comparable to those of Y-27632. Interestingly, CECs expanded in a medium supplemented with AR-13503 were significantly more proliferative in (i) untreated vs. AR-13503 (1 µM; * p < 0.05); (ii) untreated vs. AR-13503 (10 µM; *** p < 0.001); (iii) Y-27632 vs. AR-13503 (10 µM; ** p < 0.005); (iv) AR-13324 (1 µM) vs. AR-13503 (10 µM; ** p < 0.005); and (v) AR-13324 (0.1 µM) vs. AR-13503 (10 µM; * p < 0.05). Lastly, an ex vivo corneal wound healing study showed a comparable wound healing rate for the final healed area in corneas exposed to Y-27632 or AR-13324. (4) In conclusion, we were able to demonstrate that various classes of ROCKi compounds other than Y-27632 were able to exert positive effects on primary CECs, and systematic donor-match controlled comparisons revealed that the FDA-approved ROCK inhibitor, AR-13324, is a potential candidate for cellular therapeutics or as an adjunct drug in regenerative treatment for corneal endothelial diseases in humans.

Prevalence and risk factors for dry eye disease: the Sao Paulo dry eye study / Prevalência e fatores de risco para doença do olho seco: o estudo de olho seco - São Paulo

ABSTRACT Purpose: To estimate the prevalence and risk factors of dry eye disease symptoms and clinical diagnoses in Sao Paulo city, state of Sao Paulo, Brazil. Methods: A total of 582 participants over 18 years old, living in the east zone of Sao Paulo city responded to a short questionnaire. Dry eye disease was on that is defined by the presence of severe symptoms or previous clinical diagnosis of dry eye disease by an ophthalmologist. The association between dry eye disease and possible risk factors was assessed. Results: Overall dry eye disease severe symptoms and/or clinical diagnoses prevalence was calculated as 24.4% for both sexes. Women presented a higher frequency of severe symptoms of dry eye disease (16.07%) than men (8.48%; p=0.0244), as well as the composite of severe symptoms or diagnosed dry eye disease, presented by 26.86% of women and 18.18% of men (p=0.0366). In women, ages between 55 to 75 years old were associated with dry eye disease severe symptoms (OR=3.11; 95%CI 1.56-6.23, p=0.001) and diagnosed dry eye disease (OR=2.02; 95% CI 1.04-3.93, p=0.037). Hypertension was significantly associated with dry eye disease symptoms (OR=1.98; 95% CI 1.14-3.43, p=0.015) and diagnoses (OR=3.54; 95% CI 1.92-6.53, p=0.0001) in women. Eye drops use was associated with severe symptoms of dry eye disease and diagnosed dry eye disease in both women and men (p≤0.01). Conclusions: Dry eye disease prevalence in Sao Paulo city is higher in women than in men. Age and hypertension were stronger risk factors of dry eye disease for women, while eye drops use was a significant indicator of dry eye disease for both sexes.

RESUMO Objetivo Estimar a prevalência e os fatores de risco para os sintomas e o diagnóstico clínico da doença do olho seco na cidade de São Paulo, estado de São Paulo, Brasil. Métodos: Quinhentos e oitenta e dois participantes acima de 18 anos, residentes na zona leste da cidade de São Paulo responderam a um questionário de três perguntas sobre olho seco. A doença do olho seco foi definida pela presença de sintomas severos ou diagnóstico clínico prévio de doença do olho seco por um oftalmologista. A associação entre doença do olho seco e possíveis fatores de risco foi avaliada. Resultados: A prevalência de sintomas graves da doença do olho seco e/ou diagnóstico clínico foi calculada em 24,4% para ambos os sexos. O sexo feminino apresentou uma frequência maior de sintomas severos da doença do olho seco (16,07%) que o sexo masculino (8,48%; p=0,0244), assim como a associação de sintomas severos ou diagnóstico de doença do olho seco, foi de 26,86% no sexo feminino e 18,18% no sexo masculino (p=0,0366). No sexo feminino, a faixa etária entre 55 e 75 anos de idade foi associada com sintomas severos da DOS (odds ratios (OR) = 3,11; IC 95% 1,56-6,23; p=0,001) e com doença do olho seco diagnosticada (OR=2,02; IC 95% 1,04-3,93; p=0,037). Hipertensão foi associado com sintomas da doença do olho seco (OR=1,98; IC 95% 1,14-3,43; p=0,015) e diagnóstico da doença do olho seco (OR=3,54; IC 95% 1,92-6,53; p=0,001) no sexo feminino. Uso de colírios foi associado a sintomas severos e diagnóstico da doença do olho seco em ambos os sexos (p≤0,01). Conclusão: A prevalência da doença do olho seco na cidade de São Paulo é mais frequente no sexo feminino que no masculino. Idade e hipertensão foram fatores de risco maiores para doença do olho seco no sexo feminino, enquanto uso de colírios foi um indicador de doença do olho seco para ambos os sexos.

Keratoconus epidemiology: A review.

Keratoconus is an ectatic corneal disease that affects both men and women, usually at a young age. Risk factors such as eye rubbing, environmental and genetic factors contribute to its pathogenesis. Keratoconus presents a variable prevalence, being as high as 5% of the population in the Middle East. In this review, an English-language search for keratoconus epidemiology was undertaken using the PubMed database. We present and discuss the findings of the most relevant epidemiological studies on keratoconus and describe its characteristics and risk factors, correlating them with its pathogenesis.

Prevalence and risk factors for dry eye disease: the Sao Paulo dry eye study.

PURPOSE: To estimate the prevalence and risk factors of dry eye disease symptoms and clinical diagnoses in Sao Paulo city, state of Sao Paulo, Brazil. METHODS: A total of 582 participants over 18 years old, living in the east zone of Sao Paulo city responded to a short questionnaire. Dry eye disease was on that is defined by the presence of severe symptoms or previous clinical diagnosis of dry eye disease by an ophthalmologist. The association between dry eye disease and possible risk factors was assessed. RESULTS: Overall dry eye disease severe symptoms and/or clinical diagnoses prevalence was calculated as 24.4% for both sexes. Women presented a higher frequency of severe symptoms of dry eye disease (16.07%) than men (8.48%; p=0.0244), as well as the composite of severe symptoms or diagnosed dry eye disease, presented by 26.86% of women and 18.18% of men (p=0.0366). In women, ages between 55 to 75 years old were associated with dry eye disease severe symptoms (OR=3.11; 95%CI 1.56-6.23, p=0.001) and diagnosed dry eye disease (OR=2.02; 95% CI 1.04-3.93, p=0.037). Hypertension was significantly associated with dry eye disease symptoms (OR=1.98; 95% CI 1.14-3.43, p=0.015) and diagnoses (OR=3.54; 95% CI 1.92-6.53, p=0.0001) in women. Eye drops use was associated with severe symptoms of dry eye disease and diagnosed dry eye disease in both women and men (p≤0.01). CONCLUSIONS: Dry eye disease prevalence in Sao Paulo city is higher in women than in men. Age and hypertension were stronger risk factors of dry eye disease for women, while eye drops use was a significant indicator of dry eye disease for both sexes.

Bevacizumab in High-Risk Corneal Transplantation: A Pilot Multicenter Prospective Randomized Control Trial.

PURPOSE: To determine the efficacy of local (subconjunctival and topical) bevacizumab (Avastin) treatment in patients undergoing vascularized high-risk corneal transplantation. DESIGN: Pilot, prospective, randomized, double-blind, placebo-controlled clinical trial conducted at 5 clinical centers in the United States, India, and Brazil. PARTICIPANTS: Patients aged > 18 years undergoing high-risk penetrating keratoplasty, defined as corneal neovascularization (NV) in 1 or more quadrants ≥2 mm from the limbus or extension of corneal NV to the graft-host junction in a previously failed graft. METHODS: Patients were randomized to receive subconjunctival bevacizumab (2.5 mg/0.1 ml) or placebo at the time of surgery, followed by topical bevacizumab (10 mg/ml) or topical placebo, administered 4 times per day for 4 weeks. MAIN OUTCOME MEASURE: The 52-week endothelial immune rejection rate. RESULTS: Ninety-two patients were randomized to receive bevacizumab (n = 48) or control (n = 44). The 52-week endothelial rejection rate was 10% in the bevacizumab group and 19% in the control group (P = 0.20). Post hoc, extended follow-up at the lead study site showed an endothelial rejection rate of 3% in the bevacizumab group and 38% in the control group (P = 0.003). Treatment with bevacizumab was found to have a hazard ratio of 0.15 (95% confidence interval, 0.03-0.65, P = 0.01) in a post hoc Cox regression analysis. CONCLUSIONS: In patients undergoing vascularized high-risk corneal transplantation, there was no statistically significant difference in the rate of endothelial rejection at 1 year in the bevacizumab treatment group compared with the control group. This study may have been underpowered to detect a difference between treatment groups, and taken together, our data suggest that, in the current trial design, bevacizumab has a positive but not (yet) significant effect on endothelial rejection.

Posterior lamellar keratoplasty: techniques, outcomes, and recent advances.

Over the past 2 decades, posterior lamellar keratoplasty (PLK) has emerged as an alternative to penetrating keratoplasty in the treatment of corneal endothelial disorders. The reasons for this trend include the search for a safer procedure to replace diseased endothelium that provides faster and better visual rehabilitation and reduces the need for postoperative care. Different surgical techniques, surgical instruments, devices, and lasers have been introduced to overcome technical difficulties, thus improving clinical outcomes. Yet, surgeons and eye banks must address the complications and limitations that arise during the transition to these new techniques. This review discusses the most significant aspects of the evolution of PLK, including a detailed description of current techniques and the direction of future treatment for corneal endothelial disease with the use of laser-assisted surgery, bioengineered corneas, cell therapy, and new pharmacologic therapy.

Global Consensus on the Management of Limbal Stem Cell Deficiency.

PURPOSE: In recent decades, the medical and surgical treatment of limbal stem cell deficiency (LSCD) has evolved significantly through the incorporation of innovative pharmacological strategies, surgical techniques, bioengineering, and cell therapy. With such a wide variety of options, there is a need to establish a global consensus on the preferred approaches for the medical and surgical treatment of LSCD. METHODS: An international LSCD Working Group was established by the Cornea Society in 2012 and divided into subcommittees. Four face-to-face meetings, frequent email discussions, and teleconferences were conducted since then to reach agreement on a strategic plan and methods after a comprehensive literature search. A writing group drafted the current study. RESULTS: A consensus in the medical and surgical management of LSCD was reached by the Working Group. Optimization of the ocular surface by eyelid and conjunctival reconstruction, antiinflammatory therapy, dry eye and meibomian gland dysfunction treatment, minimization of ocular surface toxicity from medications, topical medications that promote epithelialization, and use of a scleral lens is considered essential before surgical treatment of LSCD. Depending on the laterality, cause, and stage of LSCD, surgical strategies including conjunctival epitheliectomy, amniotic membrane transplantation, transplantation of limbal stem cells using different techniques and sources (allogeneic vs. autologous vs. ex vivo-cultivated), transplantation of oral mucosal epithelium, and keratoprosthesis can be performed as treatment. A stepwise flowchart for use in treatment decision-making was established. CONCLUSIONS: This global consensus provides an up-to-date and comprehensive framework for the management of LSCD.

TFOS European Ambassador meeting: Unmet needs and future scientific and clinical solutions for ocular surface diseases.

The mission of the Tear Film & Ocular Surface Society (TFOS) is to advance the research, literacy, and educational aspects of the scientific field of the tear film and ocular surface. Fundamental to fulfilling this mission is the TFOS Global Ambassador program. TFOS Ambassadors are dynamic and proactive experts, who help promote TFOS initiatives, such as presenting the conclusions and recommendations of the recent TFOS DEWS II™, throughout the world. They also identify unmet needs, and propose future clinical and scientific solutions, for management of ocular surface diseases in their countries. This meeting report addresses such needs and solutions for 25 European countries, as detailed in the TFOS European Ambassador meeting in Rome, Italy, in September 2019.

Corneal crosslinking: Current protocols and clinical approach.

Members of the ASCRS Cornea Clinical Committee performed a review of the current literature on the corneal crosslinking (CXL) procedure for treating corneal ectasia. The members explored the data on the techniques currently in use and under investigation, including their advantages, safety profiles, risks, and cost analyses, compared with data on corneal transplantation. They concluded that CXL limits the progression of keratoconus, thus reducing the need for transplantation. They also found that compared with permitting the disease to progress naturally, CXL techniques carry significant and long-term cost and safety benefits, primarily by reducing the need for corneal transplantation. Studies of various CXL techniques (eg, epithelium-on treatment, changes in ultraviolet light parameters, riboflavin composition) continue with the ultimate goal of improving the procedure's safety and efficacy.

An algorithm for the preoperative diagnosis and treatment of ocular surface disorders.

Any ocular surface disease (OSD), but most commonly, dry-eye disease (DED), can reduce visual quality and quantity and adversely affect refractive measurements before keratorefractive and phacorefractive surgeries. In addition, ocular surgery can exacerbate or induce OSD, leading to worsened vision, increased symptoms, and overall dissatisfaction postoperatively. Although most respondents of the recent annual American Society of Cataract and Refractive Surgery (ASCRS) Clinical Survey recognized the importance of DED on surgical outcomes, many were unaware of the current guidelines and most were not using modern diagnostic tests and advanced treatments. To address these educational gaps, the ASCRS Cornea Clinical Committee developed a new consensus-based practical diagnostic OSD algorithm to aid surgeons in efficiently diagnosing and treating visually significant OSD before any form of refractive surgery is performed. By treating OSD preoperatively, postoperative visual outcomes and patient satisfaction can be significantly improved.

Global Consensus on Definition, Classification, Diagnosis, and Staging of Limbal Stem Cell Deficiency.

PURPOSE: Despite extensive knowledge gained over the last 3 decades regarding limbal stem cell deficiency (LSCD), the disease is not clearly defined, and there is lack of agreement on the diagnostic criteria, staging, and classification system among treating physicians and research scientists working on this field. There is therefore an unmet need to obtain global consensus on the definition, classification, diagnosis, and staging of LSCD. METHODS: A Limbal Stem Cell Working Group was first established by The Cornea Society in 2012. The Working Group was divided into subcommittees. Four face-to-face meetings, frequent email discussions, and teleconferences were conducted since then to obtain agreement on a strategic plan and methodology from all participants after a comprehensive literature search, and final agreement was reached on the definition, classification, diagnosis, and staging of LSCD. A writing group was formed to draft the current manuscript, which has been extensively revised to reflect the consensus of the Working Group. RESULTS: A consensus was reached on the definition, classification, diagnosis, and staging of LSCD. The clinical presentation and diagnostic criteria of LSCD were clarified, and a staging system of LSCD based on clinical presentation was established. CONCLUSIONS: This global consensus provides a comprehensive framework for the definition, classification, diagnosis, and staging of LSCD. The newly established criteria will aid in the correct diagnosis and formulation of an appropriate treatment for different stages of LSCD, which will facilitate a better understanding of the condition and help with clinical management, research, and clinical trials in this area.

The impact of dry eye disease treatment on patient satisfaction and quality of life: A review.

Several aspects of the quality of life (QoL) and treatment satisfaction of patients with dry eye disease (DED) may be underestimated. Ocular symptoms, which are assessed by validated patient-reported questionnaires and may include stinging, burning, itchiness, grittiness, dryness and discomfort, reduce QoL by affecting daily activities and work productivity. Self-reported symptoms do not always correlate with post-treatment improvements in clinical measures such as tear film break-up time, inflammation and osmolarity. Thus, treatments may improve clinical ocular features without improving symptoms that affect daily life. This review explores 1500 abstracts from congress presentations and peer-reviewed journals for QoL and treatment satisfaction data on the use of active lubricants, osmoprotectants, secretagogues, and immunomodulators present in topical formulations for DED treatment, and validated symptom questionnaires. Patient-reported symptoms of DED are generally improved after treatment with topical formulations for tear replacement, tear stimulation or anti-inflammatory therapy compared with baseline or a control treatment. However, more data are required to compare the performance of active ingredients. It is fundamental to diagnose patients with DED accurately, recognising the major cause behind their dry eyes. Studies are also necessary to identify how patient satisfaction and QoL may be improved through long-term use of topical preparations. We conclude that careful and thorough consideration of patient-reported symptoms should be integrated into DED management to help tailor treatment to patient needs.

Corneal Cross-Linking: Current USA Status: Report From the Cornea Society.

The initial published clinical report on riboflavin/ultraviolet A corneal cross-linking (CXL) for treatment of progressive keratoconus dates back to 2003. CXL has since then been widely used outside the United States for treatment of progressive keratoconus and post-laser in situ keratomileusis ectasia. The Food and Drug Administration (FDA) approved Avedro Inc.'s corneal cross-linking system (KXL) for treatment of patients with progressive keratoconus and post-laser in situ keratomileusis ectasia in April 2016. The procedure is not currently approved for stable keratoconus. There are 2 FDA-approved topical ophthalmic solutions for use in CXL. Riboflavin 5'-phosphate in 20% dextran ophthalmic solution 0.146% (Photrexa Viscous) and Riboflavin 5'-phosphate ophthalmic solution 0.146% (Photrexa) are intended for use with the KXL system. Photrexa Viscous is used in all CXL procedures, whereas Photrexa is indicated for use when the corneal stroma is thinner than 400 µm after completion of the Photrexa Viscous induction period. The FDA-approved procedure using the Dresden protocol (UV-A, 3 mW/cm for 30 min) induces cytologic and morphologic changes in the anterior 250 to 300 µm of the corneal stroma. It has been believed that a minimum thickness of 400 µm was necessary to protect the corneal endothelium from potential damage. The CXL procedure using the standard Dresden protocol is established as the gold standard for treatment of progressive keratoconus. CXL treatment is indicated for a list of conditions ranging from corneal ectasia to infectious keratitis. Newer protocols, treatment regimens, and expanded indications will require further refinements, investigations, and long-term studies.

TFOS DEWS II Report Executive Summary.

This article presents an Executive Summary of the conclusions and recommendations of the 10-chapter TFOS DEWS II report. The entire TFOS DEWS II report was published in the July 2017 issue of The Ocular Surface. A downloadable version of the document and additional material, including videos of diagnostic and management techniques, are available on the TFOS website: www.TearFilm.org.

Surgical techniques and adjuvants for the management of primary and recurrent pterygia.

The removal and rate of recurrence of pterygium have been discussed for years. The disorder is highly associated with environmental factors, and recurrence rates can be unacceptably high and cannot be successfully predicted. New techniques and graft preparations and postoperative management strategies are helping to reduce the recurrence rates and provide an ocular surface that is near ideal for future cataract or refractive surgery. This review discusses the advantages and disadvantages of various treatment strategies.

Supra-organization and optical anisotropies of the extracellular matrix in the amniotic membrane and limbal stroma before and after explant culture.

In this research we evaluated the supramolecular organizations and the optical anisotropical properties of the de-epithelialized human amniotic membrane and rabbit limbal stroma, before and after explant culture. Birefringence, monochromatic light spectral absorption and linear dichroism of the main extracellular matrix biopolymers, that is, the fibrillar collagens and proteoglycans, were investigated by polarized light microscopy combined with image analysis. Our results demonstrated that the culture procedure-induced stimuli altered the supra-organizational characteristics (in terms of collagens/proteoglycans spatial orientation and ordered-aggregational state) of the amniotic and limbal extracellular matrix, which led to changes in optical anisotropical properties.