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PURPOSE To assess the implementation outcomes of a new asynchronous virtual clinic, the Glaucoma Observation Clinic (GLOC), at the Singapore National Eye Center, in monitoring patients at low risk for glaucoma progression. METHODS Patients with low risk of glaucoma progression were followed up at GLOC. Visual acuity, intraocular pressure, and visual field testing or optic nerve head imaging were assessed by nurses and technicians, with virtual review of data by an ophthalmologist separately. The implementation outcomes were defined as patient satisfaction, assessed by a nurse-administered questionnaire; the rate of referrals back to the glaucoma outpatient department; the patients' journey time (minutes); the time taken for the specialist review (minutes), and the per capita manpower cost compared with current glaucoma outpatient model. RESULTS A total of 377 patients were included, with the majority being glaucoma disc suspects (n = 250, 66.4%) and primary angle closure suspects (n = 54, 14.3%). Most patients (more than 90%) reported being satisfied, and only 7.7% (n = 29) were referred back to the specialist outpatient glaucoma clinic due to possible glaucoma progression. Compared with the glaucoma outpatient clinic, the average journey time in GLOC was 50% less (59.3 mins vs 132 mins), the average time for a specialist review was 3 times faster (5.8 mins vs 19.5 mins), and the per capita manpower cost of GLOC was halved ($20.07 vs $39.78). CONCLUSIONS GLOC is a time-efficient, cost-saving, and sustainable model of care for managing patients with low risk of glaucoma progression. It was well received by patients and freed up the conventional clinic to treat patients with complex needs.

To investigate whether recently identified genetic loci for primary angle-closure glaucoma (PACG) are associated with disease severity.Case-control study.Eight hundred four PACG patients and 943 control participants of Chinese ethnicity from Singapore.The 8 PACG-associated single nucleotide polymorphisms (SNPs; rs11024102 at PLEKHA7, rs3753841 at COL11A1, rs1015213 located between PCMTD1 and ST18 on chromosome 8q, rs3816415 at EPDR1, rs1258267 at CHAT, rs736893 at GLIS3, rs7494379 at FERMT2, and rs3739821 mapping in between DPM2 and FAM102A) identified from genome-wide association studies were tested for association with disease severity using logistic regression adjusted for age and gender. A P value of 0.006 was set as significant after Bonferroni correction for testing of 8 loci. We also calculated the weighted genetic risk score (GRS) weighted by the estimated individual SNP effect size on PACG calculated as logarithm of the odds ratio (OR). Disease severity was based on the visual field mean deviation (MD) and classified as early to moderate (MD,-12 dB) and severe (MD,-20 dB).Association of PACG loci with severe disease.Of the 804 PACG patients, genotyping data were available for 768 individuals and included 436 with mild-to-moderate PACG and 206 with severe PACG. The PACG patients were significantly older (mean age, 64.3 ± 9.1 years vs. 56.4 ± 8.9 years; P0.001) and there were proportionately more women compared with control participants (58.4% vs. 49.0%; P0.001). Of the 8 loci investigated, we observed significant evidence of association with severe PACG at 1 SNP, namely rs3816415 in EPDR1 (OR, 2.03; 95% confidence interval [CI], 1.49-2.78; P = 1 × 10Our results show that EPDR1 is associated significantly with severe PACG, suggesting that it may predispose patients to more aggressive disease development. Individuals with PACG with a higher GRS were associated with a higher risk of severe PACG.

Purpose To compare the distribution of plateau iris in eyes across varying severity of primary angle closure glaucoma (PACG) using standardized ultrasound biomicroscopy (UBM) criteria. Design Cross-sectional study. Methods UBM was performed on 210 patients with PACG who had previously undergone laser peripheral iridotomy. Plateau iris was defined as the presence of all the following UBM criteria in ≥2 quadrants of the angle: anteriorly directed ciliary body, absent ciliary sulcus, iris angulation, flat iris plane, and iridotrabecular contact. Disease severity was based on the visual field mean deviation (MD) and classified as early-to-moderate (MD ≥ −12 dB), advanced (−12.01 dB to −20 dB), and severe (MD Results Of 210 subjects recruited, 23 were excluded because of poor quality UBM images. The remaining 187 patients were categorized as having early-to-moderate (n = 103), advanced (n = 38), and severe PACG (n = 46). Of these subjects, 48.1% were male, and 90.9% were of Chinese ethnicity. The overall proportion of plateau iris was 36.9%, with 32.0% (33/103) in early-to-moderate, 34.2% (13/38) in advanced, and 50% (23/46) in severe PACG (P = .03, comparing severe PACG with early-to-moderate groups). Among the severe PACG group, those with plateau iris configuration had significantly smaller anterior chamber area (P = .03) and volume (P = .01) compared with those without plateau iris. Conclusion The higher proportion of plateau iris configuration in eyes with severe PACG compared with early-to-moderate PACG suggest that this may be a contributory factor for disease severity.

Joseph Anthony Tumbocon,1,2 Tina T Wong,3– 5 Thanendthire Sangapillai,6 Yung-Chang Yen,7 Sang-Woo Park,8 Hsien Han Lim,9 Ngamkae Ruangvaravate10 1Eye Institute, St. Luke’s Medical Center, Quezon City, Philippines; 2Department of Ophthalmology, St. Luke’s Medical Center College of Medicine, Quezon City, Philippines; 3Glaucoma Department, Singapore National Eye Centre, Singapore, Singapore; 4Cataract & Comprehensive Ophthalmology Department, Singapore Eye Research Institute, Singapore, Singapore; 5Ophthalmology & Visual Sciences, Duke-NUS Graduate Medical School, Singapore, Singapore; 6Tun Hussein Onn National Eye Hospital, Petaling Jaya, Selangor, Malaysia; 7Department of Ophthalmology, Chi Mei Medical Center, Tainan City, Taiwan; 8Department of Ophthalmology, Chonnam National University Medical School and Hospital, Gwangju, South Korea; 9Eye Centre, Sunway Medical Centre, Petaling Jaya, Selangor, Malaysia; 10Department of Ophthalmology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, ThailandCorrespondence: Joseph Anthony Tumbocon, Eye Institute, St. Luke’s Medical Center, 279 E Rodriguez Sr. Ave, Quezon City, Metro Manila, 1112, Philippines, Email jatumbocon@gmail.comBackground: Preserved prostaglandin analogs (PGAs) have been linked to ocular surface disease (OSD). While the benefits of preservative-low (PL) or -free (PF) Tafluprost (Santen Inc., Japan) were reported in real-world studies in Western countries, this is the first study in Asia to assess the effectiveness and safety of switching from preserved PGA to PL or PF-Tafluprost.Methods: We conducted a meta-analysis on studies that included adults (> 18 years of age) with a Corneal Fluorescein Staining Score (CFS) > 1. These individuals had switched to PL or PF-tafluprost after using a preserved PGA therapy for at least 3 months for glaucoma and were identified from Santen’s tafluprost study database. A total of six studies from South Korea, Philippines, Malaysia, Singapore, Thailand, Taiwan were pooled for analysis.Results: An intraocular pressure (IOP) reduction of 5.9% (0.91 mmHg) was seen in 265 patients. However, this result was not statistically significant (95% CI: − 3.64, 1.81; Figure 1). Among 132 patients, a 47.9% reduction in the CFS (95% CI: − 3.65, − 1.91) was seen. Tear film break-up (n=183) significantly increased by 1.06 seconds (95% CI: 0.65, 1.47). In 88 patients, the bulbar conjunctiva score decreased by − 0.46 (95% CI: − 0.81, − 0.10) and palpebral conjunctiva score decreased by − 0.42 (95% CI: − 0.67– 0.17). One or more new adverse reactions were reported in 3% of the individuals after switching.Conclusion: Tafluprost IOP reduction is comparable to other PGAs, with significant improvements in the ocular surface and minimal adverse reactions which were already previously reported.Keywords: ocular surface disease, glaucoma, preserved prostaglandins, tafluprost, PGA, preservative-free, preservative-low, Asian population, meta analysis

Purpose Trabeculectomy surgery could affect ocular surface disease (OSD) in several ways, through cessation of long term glaucoma eyedrops, exposure to operative mitomycin C and post-operative eyedrops including corticosteroids and aminoglycosides and reduction in eyelid hygiene measures. Previously we showed the relevance of tear lipid mediators (also referred oxylipins) in OSD. Here, we aim to evaluate changes of these lipids in a post-trabeculectomy cohort. Methods Patients undergoing trabeculectomy were prospectively evaluated and had tear collected using Schirmer's strips, preoperatively and postoperatively at 0.5, 1.0 and 3.0 years. Lipid mediators were analyzed using liquid chromatography mass spectrometry. Results The normalized concentrations of 40 lipid mediators were between 0.1 and 8.0 ng/mL, whereas docosahexaenoic acid (DHA), Arachidonic acid (AA) and eicosapentaenoic acid (EPA) ranged up to a few hundred ng/mL. The concentrations of lipid mediators, except DHA, EPA, and thromboxane (TXB1), showed reduction after surgery. At the last visit, these lipids were significantly reduced by 1/3 to ½, compared to pre-operative values: 8-HETE, 15-HETE, 15-oxoETE, 11-HDoHE, 17-HDoHE, and 20-OH-LTB4. To examine collective changes of lipids, clustering analysis revealed 10 groups of lipids consistent with known metabolic pathways. Results An increase in the level of 2,3-dinor-8-isoPGF2α between 0 and 0.5 year was associated with inferior corneal staining at 0.5 year. In 14 patients who required post-operative needling, six lipid mediators were found to be significantly higher at 1.0 year compared to non-needled patients. Conclusions In this 3-years study, trabeculectomy reduced the tear level of pro-inflammatory lipid mediators. Patients who required needling of the bleb to maintain surgical success may have a chronic underlying inflammatory process associated with fibrosis.

Glaucoma filtration surgery plays an important role in achieving intraocular pressure (IOP) reduction in patients who have high IOP despite maximum medical therapy. Preclinical experimental models of glaucoma filtration surgery contribute a great deal to our knowledge of the wound healing processes that predispose to scarring and may lead to poor outcomes. However, this research needs to be interpreted in the light of the specific study design, animal model and methods used. We review the existing literature addressing various models of experimental glaucoma filtration surgery, discuss the considerations in assessing these models and describe future steps in evaluating potential therapeutics and bleb characteristics that could impact translational research in this field.

Background Hydroxyapatite (HA) [Ca5(PO4)3(OH)] is a naturally occurring calcium phosphate which makes up 60-70% of the dry weight of human bones. Nano-scale HA particles are increasingly being used as carriers for controlled and targeted delivery of bioactive agents like drugs, proteins, and nucleic acids due to their high porosity, negative charge, and biodegradability. Purpose Although much effort has been devoted to understanding the delivery kinetics and effects of the payloads in such carriers, a thorough understanding of the influence of the carriers themselves is lacking. Methods HA particles (300 µg/mL) were administered to primary human dermal fibroblasts (HDFs). The uptake and intracellular localization of the particles were determined by flow cytometry, confocal imaging, and transmission electron microscopy (TEM). Immunological assays and PCR were performed to determine the levels of pro-inflammatory cytokines and collagens in cell lysates and media supernatant. Results The current study explores the effects of poly-dispersed HA particles on primary HDFs as a model system. The majority of the particles were determined to range between 150 and 200 nm in diameter. Upon exposure to HA suspensions, primary HDFs internalized the particles by endocytosis within 6 hours of exposure, showing maximum uptake at 72 hours following which the particles were exocytosed by 168 hours. This correlated to reduced secretion of various pro-inflammatory and pro-collagenic cytokines. Biochemical analysis further revealed a reduction in Type I collagen expression and secretion. Conclusion HA particles have an immune-modulatory effect on dermal fibroblasts and reduce collagen production, which may impact the integrity of the extracellular matrix (ECM). This study demonstrates the need to consider the secondary effects of particulate carriers like HA, beyond basic cytotoxicity, in the specific tissue environment where the intended function is to be realized.

The human eye has a unique immune architecture and behavior. While the conjunctiva is known to have a well-defined lymphatic drainage system, the cornea, sclera, and uveal tissues were historically considered "alymphatic" and thought to be immune privileged. The very fact that the aqueous outflow channels carry a clear fluid (aqueous humor) along the outflow pathway makes it hard to ignore its lymphatic-like characteristics. The development of novel lymphatic lineage markers and expression of these markers in aqueous outflow channels and improved imaging capabilities has sparked a renewed interest in the study of ocular lymphatics. Ophthalmic lymphatic research has had a directional shift over the last decade, offering an exciting new physiological platform that needs further in-depth understanding. The evidence of a presence of distinct lymphatic channels in the human ciliary body is gaining significant traction. The uveolymphatic pathway is an alternative new route for aqueous outflow and adds a new dimension to pathophysiology and management of glaucoma. Developing novel animal models, markers, and non-invasive imaging tools to delineate the core anatomical structure and physiological functions may help pave some crucial pathways to understand disease pathophysiology and help develop novel targeted therapeutic approaches for glaucoma. Agency for Science, Technology and Research (A*STAR) National Medical Research Council (NMRC) National Research Foundation (NRF) Funding for this work came from the National Institutes of Health, Bethesda, MD (grant numbers R01EY030501 [ASH] and R21EY026260 [YKH]); the Duke-NUS Medical School (DukeNUS-KP(Coll)/2018/0009A), the National Medical Research Council (CG/C010A/2017, OFIRG/0048/2017, OFLCG/004c/2018, and TA/MOH-000249-00/2018), National Research Foundation Singapore, A∗STAR (A20H4b0141), the Singapore Eye Research Institute & Nanyang Technological University (SERI-NTU Advanced Ocular Engineering (STANCE) Program), the SERI-Lee Foundation (LF1019-1) Singapore, Glaucoma Research Foundation Shaffer Grant [ASH], and an unrestricted grant from Research to Prevent Blindness [UCLA] (New York, NY).

Purpose This study aimed to compare the effectiveness of combination therapy consisting of low-dose mitomycin C (MMC) and valproic acid (VPA) against high-dose MMC for improving the scar phenotype in minimally invasive glaucoma surgery (MIGS). Methods A rabbit model of MIGS incorporating the PreserFlo MicroShunt was treated with high (0.4 mg/mL) or low (0.1 mg/mL) doses of MMC or with combination therapy consisting of low-dose (0.1 mg/mL) MMC and VPA. Operated eyes were examined by live ocular imaging, histochemical evaluation, multiphoton quantitation of collagen characteristics, and molecular analyses. Results Although high-dose MMC obliterated the vasculature, combination therapy vastly improved the postoperative tissue morphology by maintaining the vasculature without increased vascularization. Combination therapy also altered collagen morphology and reduced encapsulation of the MicroShunt distal end, which remained at risk with MMC treatment alone. Multiphoton quantitation indicated that the combination therapy significantly reduced collagen density and fiber dimensions compared with monotherapy. At the molecular level, combination therapy significantly reduced Vegfa, Vegfc, and Vegfd expression and inhibited Col1a1 upregulation from baseline levels, all of which low-dose MMC alone was unable to achieve. Notably, COL1A1 protein levels appeared more consistently suppressed by combination therapy compared with high-dose MMC alone. Conclusions Compared with high-dose MMC, combination therapy was less toxic by sparing the vasculature and potentially more effective in reducing scarring via the regulation of collagen content and organization. Translational Relevance VPA may be combined with low-dose MMC to replace high-dose MMC to deliver safe and effective anti-scarring outcomes.

To assess the implementation outcomes of a new asynchronous virtual clinic, the Glaucoma Observation Clinic (GLOC), at the Singapore National Eye Center, in monitoring patients at low risk for glaucoma progression.Patients with low risk of glaucoma progression were followed up at GLOC. Visual acuity, intraocular pressure, and visual field testing or optic nerve head imaging were assessed by nurses and technicians, with virtual review of data by an ophthalmologist separately. The implementation outcomes were defined as patient satisfaction, assessed by a nurse-administered questionnaire; the rate of referrals back to the glaucoma outpatient department; the patients' journey time (minutes); the time taken for the specialist review (minutes), and the per capita manpower cost compared with current glaucoma outpatient model.A total of 377 patients were included, with the majority being glaucoma disc suspects (n = 250, 66.4%) and primary angle closure suspects (n = 54, 14.3%). Most patients (more than 90%) reported being satisfied, and only 7.7% (n = 29) were referred back to the specialist outpatient glaucoma clinic due to possible glaucoma progression. Compared with the glaucoma outpatient clinic, the average journey time in GLOC was 50% less (59.3 mins vs 132 mins), the average time for a specialist review was 3 times faster (5.8 mins vs 19.5 mins), and the per capita manpower cost of GLOC was halved ($20.07 vs $39.78).GLOC is a time-efficient, cost-saving, and sustainable model of care for managing patients with low risk of glaucoma progression. It was well received by patients and freed up the conventional clinic to treat patients with complex needs.

Valproic acid (VPA), widely used for the treatment of neurological disorders, has anti-fibrotic activity by reducing collagen production in the postoperative conjunctiva. In this study, we investigated the capacity of VPA to modulate the postoperative collagen architecture. Histochemical examination revealed that VPA treatment was associated with the formation of thinner collagen fibers in the postoperative days 7 and 14 scars. At the micrometer scale, measurements by quantitative multiphoton microscopy indicated that VPA reduced mean collagen fiber thickness by 1.25-fold. At the nanometer scale, collagen fibril thickness and diameter measured by transmission electron microscopy were decreased by 1.08- and 1.20-fold, respectively. Moreover, delicate filamentous structures in random aggregates or closely associated with collagen fibrils were frequently observed in VPA-treated tissue. At the molecular level, VPA reduced Col1a1 but induced Matn2, Matn3, and Matn4 in the postoperative day 7 conjunctival tissue. Elevation of matrilin protein expression induced by VPA was sustained till at least postoperative day 14. In primary conjunctival fibroblasts, Matn2 expression was resistant to both VPA and TGF-β2, Matn3 was sensitive to both VPA and TGF-β2 individually and synergistically, while Matn4 was modulable by VPA but not TGF-β2. MATN2, MATN3, and MATN4 localized in close association with COL1A1 in the postoperative conjunctiva. These data indicate that VPA has the capacity to reduce collagen fiber thickness and potentially collagen assembly, in association with matrilin upregulation. These properties suggest potential VPA application for the prevention of fibrotic progression in the postoperative conjunctiva. KEY MESSAGES: VPA reduces collagen fiber and fibril thickness in the postoperative scar. VPA disrupts collagen fiber assembly in conjunctival wound healing. VPA induces MATN2, MATN3, and MATN4 in the postoperative scar.

Purpose: This study investigated the involvement of NF-kB in regulating postoperative conjunctival inflammation.Methods: Experimental surgery was performed as described for the mouse model of conjunctival scarring. Expression of NF-κB in postoperative conjunctival tissues or conjunctival fibroblasts were assessed by real-time PCR, immunoblotting and immunofluorescence analyses. Downregulation of RelB was achieved using small interfering RNA. Cellular cytokine secretion was determined using multiplex cytokine assay.Results: RelB was the most highly induced member of the NF-kB family on day 2 post-surgery. Elevated RelB may be found associated with vimentin-positive cells and fibroblasts in vivo and in vitro. In conjunctival fibroblasts, RelB may be induced by TNF-α but not TGF-β2 while its silencing caused selective induction of CCL2 secretion by both basal and TNF-α-stimulated fibroblasts.Conclusions: High RelB induction in the inflammatory phase and the selective modulation of CCL2 suggest a specific anti-inflammatory role for RelB in the postoperative conjunctiva.

Background: Poor adherence to medications is a global public health concern with substantial health and cost implications, especially for chronic conditions. In the USA, poor adherence is estimated to cause 125,000 deaths and cost $US100 Billion annually. The most successful adherence-promoting strategies that have been identified so far have moderate effect, are relatively costly, and raise availability, feasibility, and/or scalability issues. Objective: The main objective of SIGMA (Study on Incentives for Glaucoma Medication Adherence) was to measure the effectiveness on medication adherence of a novel incentive strategy based on behavioral economics that we refer to as adherence-contingent rebates. These rebates offered patients a near-term benefit while leveraging loss aversion and regret nd increasing the salience of adherence. Methods: IGMA is a 6-month randomized, controlled, open-label, single-center superiority trial with two parallel arms. total of 100 non-adherent glaucoma patients from the Singapore National Eye Centre were randomized into intervention (adherence-contingent rebates) and usual care (no rebates) arms in a 1:1 ratio. The primary outcome was the mean change from baseline in percentage of adherent days at Month 6. The trial registration number is NCT02271269 and a detailed study protocol has been published elsewhere. Findings: We found that participants who were offered adherence-contingent rebates were adherent to all their edications on 73.1% of the days after 6 months, which is 12.2 percentage points (p = 0.027) higher than in those not receiving the rebates after controlling for baseline differences. This better behavioral outcome was achieved by rebates averaging 8.07 Singapore dollars ($US5.94 as of 2 November 2017) per month during the intervention period. Conclusion: This study shows that simultaneously leveraging several insights from behavioral economics can significantly improve medication adherence rates. The relatively low cost of the rebates and significant health and cost implications of medication non-adherence suggest that this strategy has the potential to cost-effectively improve health outcomes for many conditions.

Inhibitors of vascular endothelial growth factors are used to treat a myriad of retinal conditions, including exudative age-related macular degeneration (AMD), diabetic macular oedema (DME) and diabetic retinopathy. Although effective, long-term efficacy is limited by the need for frequent and invasive intravitreal injections. The quest for sustained action therapeutics that can be delivered to target tissue in the least invasive manner is an arduous endeavour that has ended in premature failure for several technologies in Phase II or III trials. Nevertheless, there have been promising preclinical studies, and more are on the horizon: port delivery systems for the treatment of exudative AMD have entered Phase III trials and a wide array of preclinical studies have demonstrated the potential for nanoparticles, such as liposomes, dendrimers and cell penetrating peptides to deliver therapeutics into the posterior segment via minimally invasive routes. In this review, we discuss the challenges posed by ocular barriers for drug penetration and present the recent advancements of the most pertinent drug delivery platforms with a focus on the treatment of exudative AMD and DME.

To investigate the association of peripheral anterior synechiae (PAS) with intraocular pressure (IOP) and anterior-segment parameters in subjects with primary angle-closure glaucoma (PACG). A total of 267 subjects with PACG were recruited and underwent gonioscopy and anterior-segment optical coherence tomography (ASOCT). Customized software was used to measure ASOCT parameters, including angle opening distance (AOD750) and trabecular-iris-space-area (TISA750) at 750 µm from the scleral spur, anterior chamber depth, width, area and volume (ACD, ACW, ACA, ACV), iris thickness (IT750), iris area (IAREA), and lens vault (LV). Presenting IOP was defined as the first IOP reading before the initiation of IOP-lowering treatment. The mean age of the 267 subjects was 67.0 ± 8.9 years, 140 (52.4%) were male, and 246 (92.1%) were of Chinese ethnicity. PAS was present in 122 (45.7%) subjects, and was most frequently found in the superior quadrant (79.5%). Subjects with PAS had greater presenting IOP (28.7 ± 12.9 vs 22.4 ± 9.7 mmHg, p

PurposeTo determine the effect of valproic acid (VPA) on bleb morphology and scar characteristics in a rabbit model of minimally invasive glaucoma surgery (MIGS).MethodsNine New Zealand white rabbits were subjected to MIGS with intraoperative implantation of the PreserFlo MicroShunt. Rabbits were then administered with subconjunctival injections of phosphate buffered saline (PBS) (n=4) or with VPA (n=5). Bleb morphology was examined by slit-lamp biomicroscopy and in vivo confocal microscopy. Postoperative day 28 tissues were examined by immunohistochemical evaluation and label-free multiphoton microscopy to visualise the collagen matrix, by terminal deoxynucleotidyl transferase dUTP nick-end labelling assay and immunofluorescent labelling for Ki67 expression to detect apoptosis and cell growth, and by real-time quantitative PCR to measure Col1a1, Fn, and Smad6 transcript expression.ResultsVPA-treated blebs were detectable on day 28, while the PBS-treated blebs were not detectable by day 14. VPA-treated blebs were diffuse, extended posteriorly with near normal conjunctival vascularity and featured a combination of reticular/blurred stromal pattern with evidence of relatively large stromal cysts. Instead of the deposition of thick, disorganised collagen fibres characteristic of the PBS bleb, the VPA bleb contained conspicuously thinner collagen fibres which were associated with similarly thinner fibronectin fibres. In corroboration, Col1a1 and Fn mRNA expression was reduced in the VPA blebs, while increased Smad6 expression implicated the disruption of the transforming growth factor beta pathway. Apoptosis and cell growth profiles appeared similar with both treatments.ConclusionsThe results support the application of VPA to enhance bleb morphology associated with good bleb function in MIGS with no apparent cytotoxicity.

Allergic conjunctivitis is one of the most common external eye diseases and the prevalence has been increasing. The mainstay of treatment is topical eye drops. However, low bioavailability, low ocular drug penetration, transient resident time on the ocular surface due to tear turnover, frequent topical applications and dependence on patient compliance, are the main drawbacks associated with topical administration. Nanotechnology-based medicine has emerged to circumvent these limitations, by encapsulating the drugs and preventing them from degradation and therefore providing sustained and controlled release. Using a nanotechnology-based approach to load the drug is particularly useful for the delivery of hydrophobic drugs such as immunomodulatory agents, which are commonly used in allergic conjunctival diseases. In this review, different nanotechnology-based drug delivery systems, including nanoemulsions, liposomes, nanomicelles, nanosuspension, polymeric and lipid nanoparticles, and their potential ophthalmic applications, as well as advantages and disadvantages, are discussed. We also summarize the results of present studies on the loading of immunomodulators or nonsteroidal anti-inflammatory drugs to nano-scaled drug delivery systems. For future potential clinical use, research should focus on the optimization of drug delivery designs that provide adequate and effective doses with safe and satisfactory pharmacokinetic and pharmaco-toxic profiles. Published version

Glaucoma surgeries, such as trabeculectomy, are performed to lower intraocular pressure to reduce risk of vision loss. These surgeries create a new passage in the eye that reroutes the aqueous humor outflow to the subconjunctival space, where the fluid is presumably absorbed by the conjunctival lymphatics. Here, we characterized the development and function of the ocular lymphatics using transgenic lymphatic reporter mice and rats. We found that the limbal and conjunctival lymphatic networks are progressively formed from a primary lymphatic vessel that grows from the nasal-side medial canthus region at birth. This primary lymphatic vessel immediately branches out, invades the limbus and conjunctiva, and bidirectionally encircles the cornea. As a result, the distribution of the ocular lymphatics is significantly polarized toward the nasal side, and the limbal lymphatics are directly connected to the conjunctival lymphatics. New lymphatic sprouts are produced mainly from the nasal-side limbal lymphatics, posing the nasal side of the eye as more responsive to fluid drainage and inflammatory stimuli. Consistent with this polarized distribution of the ocular lymphatics, a higher drainage efficiency was observed in the nasal side than the temporal side of the eye when injected with a fluorescent tracer. In contrast, blood vessels are evenly distributed at the anterior surface of the eyes. Also, we found that these distinct vascular distribution patterns were conserved in human eyes. Together, our study demonstrated that the ocular surface lymphatics are more densely present in the nasal side and uncovered the potential clinical benefits in selecting the nasal side as a glaucoma surgery site to improve fluid drainage.

Small interfering RNA (siRNA) therapy is a promising epigenetic silencing strategy. However, its widespread adoption has been severely impeded by its ineffective delivery into the cellular environment. Here, a biocompatible injectable gelatin-based hydrogel with positive-charge tuned surface charge is presented as an effective platform for siRNA protection and delivery. We demonstrate a two-step synthesis of a gelatin-tyramine (Gtn-Tyr) hydrogel with simultaneous charge tunability and crosslinking ability. We discuss how different physiochemical properties of the hydrogel interact with siSPARC (siRNA for secreted protein, acidic and rich in cysteine), and study the positive-charge tuned gelatin hydrogel as an effective delivery platform for siSPARC in anti-fibrotic treatment. Through in vitro studies using mouse tenon fibroblasts, the positive-charge tuned Gtn-Tyr hydrogel shows sustained siSPARC cellular internalization and effective SPARC silencing with excellent biocompatibility. Similarly, the same hydrogel platform delivering siSPARC in an in vivo assessment employing a rabbit model shows an effective reduction in subconjunctival scarring in post glaucoma filtration surgery, and is non-cytotoxic compared to a commonly used anti-scarring agent, mitomycin-C. Overall, the current siRNA delivery strategy involving the positive-charge tuned gelatin hydrogel shows effective delivery of gene silencing siSPARC for anti-fibrotic treatment. The current charge tunable hydrogel delivery system is simple to fabricate and highly scalable. We believe this delivery platform has strong translational potential for effective siRNA delivery and epigenetic silencing therapy. Ministry of Education (MOE) Nanyang Technological University National Medical Research Council (NMRC) National Research Foundation (NRF) Published version This research was supported by an NTU internal grant (M4081851), Singapore Ministry of Education NTUitive Gap Fund (NGF-2019-07-004) and the Singapore National Research Foundation under its Translational and Clinical Research (TCR) Programme (NMRC/TCR/008-SERI/2013). Animal studies were partially funded by the SERI core grant (NMRC/CG/015/2013). We would also like to acknowledge Nyein Chan Lwin from SingHealth Experimental Medicine Centre (SEMC), Singapore for assisting the in vivo studies.

The purpose of this study is to evaluate outflow pathways from subconjunctival blebs and to identify their identity. Post-mortem porcine (n = 20), human (n = 1), and bovine (n = 1) eyes were acquired, and tracers (fluorescein, indocyanine green, or fixable/fluorescent dextrans) were injected into the subconjunctival space to create raised blebs where outflow pathways were visualized qualitatively and quantitatively. Rodents with fluorescent reporter transgenes were imaged for structural comparison. Concurrent optical coherence tomography (OCT) was obtained to study the structural nature of these pathways. Using fixable/fluorescent dextrans, tracers were trapped to the bleb outflow pathway lumen walls for histological visualization and molecular identification using immunofluorescence against lymphatic and blood vessel markers. Bleb outflow pathways could be observed using all tracers in all species. Quantitative analysis showed that the nasal quadrant had more bleb-related outflow pathways compared to the temporal quadrant (nasal: 1.9±0.3 pathways vs. temporal: 0.7±0.2 pathways; p = 0.003). However, not all blebs resulted in an outflow pathway (0-pathways = 18.2%; 1-pathway = 36.4%; 2-pathways = 38.6%; and 3-pathways = 6.8%). Outflow signal was validated as true luminal pathways using optical coherence tomography and histology. Bicuspid valves were identified in the direction of flow in porcine eyes. Immunofluorescence of labeled pathways demonstrated a lymphatic (Prox-1 and podoplanin) but not a blood vessel (CD31) identity. Therefore, subconjunctival bleb outflow occurs in discrete luminal pathways. They are lymphatic as assessed by structural identification of valves and molecular identification of lymphatic markers. Better understanding of lymphatic outflow may lead to improved eye care for glaucoma surgery and ocular drug delivery.

Importance Evidence-based guidelines are essential for glaucoma screening to work effectively. Background To derive a vertical cup-to-disc ratio (VCDR) cut-off for glaucoma screening in a multi-ethnic Asian population. Design The Singapore Epidemiology of Eye Diseases (SEED) study is a population-based study conducted from 2004 to 2011 in a single tertiary care research institute. Participants SEED comprised of 10 033 Chinese, Malay and Indian adults aged ≥40 (response rate 75.6%). After excluding participants with a history of glaucoma medication or surgery, 9673 participants were included for analysis. Methods A systematic eye examination, which included applanation tonometry, visual field testing, gonioscopy, and dilated fundus examination was conducted. Main outcome measure Diagnosis of glaucoma. Results The distribution of VCDR and VCDR asymmetry were relatively homogenous in this multi-ethnic Asian population, with a 97.fifth percentile value of 0.67 and 0.17 respectively. In the absence of more definite signs of glaucoma, VCDR ≥0.60 and VCDR asymmetry ≥0.20 provided the best balance between sensitivity (95.1%) and specificity (90.9%) in detecting glaucoma. For larger optic disc (≥2.0 mm), VCDR ≥0.65 with VCDR asymmetry ≥0.20 provided the best balance between sensitivity (84.8%) and specificity (93.2%). Conclusion and relevance Overall, VCDR ≥0.60 with VCDR ≥0.20 asymmetry provides a good balance between sensitivity and specificity in detecting glaucoma. For larger optic disc, VCDR ≥0.65 should be considered instead to mitigate against false-referrals due to larger physiological disc cupping. Our findings may act as a reference to populations with similar VCDR distribution.

Valproic acid (VPA) is a histone deacetylase inhibitor used clinically for neurological disorders. It is also potentially useful as anti-fibrotic therapy as it reduced collagen deposition in the post-operative conjunctiva. In this study, we further evaluated the effects of VPA on post-operative inflammation using the mouse model of conjunctival scarring. VPA, injected into the subconjunctiva immediately after surgery, did not cause any adverse tissue response when examined by live microscopy and produced an apparent reduction of proinflammatory and proangiogenic markers in immunohistological examinations. In-depth analyses of the treated operated tissues revealed that VPA selectively inhibited the CD45highF4/80low macrophage subset as well as the production of specific proinflammatory cytokines/ chemokines, including CXCL1, IL-5, IL-6, and IL-10 which were reduced by ≥ 2.0-fold. VPA also specifically reduced tissue NF-кB2 p100 protein by mean 3.87-fold. On conjunctival fibroblasts, VPA treatment resulted in decreased secretion of specific cytokines, including CCL2, VEGF-A, and IL-15. In the presence of TNF-α, VPA inhibited the induction of specific cytokines/chemokines, notably CCL5 and VEGF-A, as well as NF-кB2 p100. In corroboration, VPA suppressed TNF-α stimulation of NF-кB reporter transcription by 1.51-fold. These data indicate that VPA can modulate both proinflammatory cellular and molecular targets in a selective manner and may therefore attenuate surgery-induced conjunctival inflammation. These and previous findings suggest that, by suppressing key mediators of both inflammation and fibrosis, VPA is a useful therapeutic for improving surgical outcome involving the conjunctiva. Key messages VPA inhibited recruitment of a CD45highF4/80low macrophage subset. VPA reduced chemokine and cytokine levels in treated tissues.VPA selectively suppressed tissue NF-кB2 p100 levels.VPA suppressed TNF-α induction of chemokines, cytokines and NF-кB2 p100 expression.VPA suppressed TNF-α stimulation of NF-кB reporter.

Background: The advent of therapeutic proteins highlights the need for delivery systems that protect and extend the duration of its action. Ranibizumab-VEGF is one such drug used for treating wet AMD. This paper describes a facile method to sustain bioactive ranibizumab release from PLGA-based particles. Methods: Two emulsion techniques were explored namely: water-in-oil-in-water (WOW) and solid-in-oil-in-water (SOW) emulsion. The bioactivity of ranibizumab was evaluated by comparing its binding capability to VEGF, measured with ELISA to total protein measured by microBCA. Results: During the emulsion process, contact of ranibizumab with the water-oil interface is the main destabilizing factor and this can be prevented with the use of amphiphilic PVA and solid-state protein in WOW and SOW emulsion respectively. In vitro release of the ranibizumab-loaded particles indicated that a 15-day release could be achieved with SOW particles while the WOW particles generally suffered from a burst release. Released ranibizumab was capable of inhibiting endothelial cell growth indicating its retention of bioactivity. The suppression of burst release from the SOW particles was attributed to the relatively smooth surface morphology of the SOW microparticles. Conclusions: The use of SOW encapsulation in modulating ranibizumab release while maintaining their bioactivity has been highlighted. MOE (Min. of Education, S’pore)

BackgroundTo develop targeted antifibrotic therapy for glaucoma filtration surgery; this study determines the effectiveness of small interfering RNA (siRNA) to reduce in vivo secreted protein acidic and rich in cysteine (SPARC) expression using the mouse model of conjunctival scarring.MethodsExperimental surgery was performed as described for the mouse model of conjunctival scarring. Scrambled (siScram) or Sparc (siSparc) siRNAs, loaded on layer-by-layer (LbL) nanoparticles, were injected into the conjunctiva immediately after surgery. Expression ofSparc,Col1a1,Fn1andMmp14was measured by real-time PCR and immunoblotting on days 7 and 14 postsurgery. Live imaging of the operated eyes was performed using slit lamp, anterior segment-optical coherence tomography and confocal microscopy. Tissue pathology was evaluated by histochemical and immunofluorescent analyses of operated conjunctival cryosections. Tissue apoptosis was quantitated by annexin V assay.Results siSparc, delivered via expanded LbL nanoparticles, significantly inhibitedSparctranscription in both day 7 (2.04-fold) and day 14 (1.39-fold) treated tissues.Sparcsuppression on day 7 was associated with a significant reduction ofCol1a1(2.52-fold),Fn1(2.89-fold) andMmp14(2.23-fold) mRNAs. At the protein level, both SPARC and collagen 1A1 (COL1A1) were significantly reduced at both time points with siSparc treatment. Nanoparticles were visualised within cell-like structures by confocal microscopy, while overt tissue response or apoptosis was not observed.Conclusions SPARC targeted therapy effectively reduced both SPARC and collagen production in the operated mouse conjunctiva. This proof-of-concept study suggests that targeted treatment of fibrosis in glaucoma surgery is safe and feasible, with the potential to extend to a range of potential genes associated with fibrosis.

Importance Glaucoma treatment has often been associated with adverse side‐effects from preservatives that are included in the used eye drops. Background To evaluate changes in the ocular surface and the presence of prostaglandin‐induced corneal disorders after being switched from latanoprost 0.005% to low preservative tafluprost 0.0015% ophthalmic solution. Design Single centre, prospective study. Participants Patients with primary open‐angle glaucoma or ocular hypertension that had received treatment with once daily latanoprost 0.005% ophthalmic solution for control of intraocular pressure (IOP) for 3 months, with a score of above 1 on the National Eye Institute (NEI) ocular surface staining scale. Methods Following the ≥3 month latanoprost treatment period, patients were switched to once daily low preservative tafluprost 0.0015% ophthalmic solution. Patients were followed for a minimum of 3 months. Main Outcome Measures Ocular surface changes were assessed by fluorescein staining score (NEI scale). Additional evaluations included tear break‐up time, hyperaemia score, subjective symptoms, changes in intraocular pressure and presence of adverse reactions. Results Out of 59 patients enrolled, 51 were included in the final analysis. Fluorescein staining scores at baseline, prior to treatment switch, were 6.9 ± 3.1 and 3.3 ± 2.7 at the end of the study period (change in scores was −3.6 ± 2.2 [P < 0.001]). At last follow‐up, significant improvements were observed in tear break‐up time, hyperaemia score and subjective symptoms (all P < 0.05). Conclusions and Relevance The clinical signs of ocular surface disease and subjective symptoms of dry eyes improved following the switch to low preservative tafluprost and demonstrated comparable IOP lowering effectiveness.

To investigate whether newly identified genetic loci for primary angle-closure glaucoma (PACG) are associated with early stage angle-closure disease defined as primary angle closure suspect (PACS).Case-control study.A total of 1397 PACS patients and 943 controls of Chinese ethnicity from Singapore and 604 PACS patients and 287 controls of Indian ethnicity.The 8 PACG single nucleotide polymorphisms (SNPs; rs11024102 at PLEKHA7, rs3753841 at COL11A1, rs1015213 located between PCMTD1 and ST18 son chromosome 8q, rs3816415 at EPDR1, rs1258267 at CHAT, rs736893 at GLIS3, rs7494379 at FERMT2, and rs3739821 mapping in between DPM2 and FAM102A) were genotyped by Taqman assays. The association between SNP genotypes and PACS status was measured using logistic regression. A P value of 0.006 was set to account for the testing of 8 genetic loci using a Bonferroni correction. A meta-analysis was conducted to calculate the overall P value and accompanying per-allele odds ratios for each SNP analyzed.Association of PACG loci with PACS status.The PACS patients were significantly older in both cohorts (Chinese, P0.001; Indian, P = 0.002), and there were also more women (P0.001, both Chinese and Indian cohorts). In the Chinese cohort, significant evidence of association was noted at 3 SNPs: rs1015213 [A] in PCMTD1-ST18 (odds ratio [OR], 2.36; 95% confidence interval [CI], 1.36-4.11; P = 0.002), rs3816415 [A] in EPDR1 (OR, 1.49; 95% CI, 1.19-1.85; P 0.001), and rs3739821 [G] in DPM2-FAM102A (OR, 1.40; 95% CI, 1.18-1.65; P0.001). Only PCMTD1-ST-18 was replicated modestly in the Indian population (P = 0.056). Meta-analysis showed significant evidence of association for PCMTD1-ST-18 (OR, 1.55; 95% CI, 1.18-2.04; P = 0.002) and DPM2-FAM102A (OR, 1.27; 95% CI, 1.12-1.45; P = 0.0002).In this study, 2 of 8 PACG-associated loci were associated significantly with PACS status, the earliest stage in the angle-closure glaucoma disease course. The association of these PACG loci with PACS status suggests that these loci may confer susceptibility to a narrow angle configuration.

Ocular anterior segment inflammation is a medical problem that is seen in cases of cataract surgery and non-infectious anterior uveitis. Inadequately treated anterior segment inflammation can lead to sight-threatening conditions such as corneal oedema, glaucoma and cystoid macular oedema. The mainstay of treatment for anterior segment inflammation is topical steroid eye-drops. However, several drawbacks limit the critical value of this treatment, including low bioavailability, poor patient compliance, relatively difficult administration manner and risk of blurring of vision and ocular irritation. A drug delivery system (DDS) that can provide increased bioavailability and sustained delivery while being specifically targeted towards inflamed ocular tissue can potentially replace daily eye-drops as the gold standard for management of anterior segment inflammation. The various DDS for anti-inflammatory drugs for the treatment of anterior segment inflammation are listed and summarised in this review, with a focus on commercially available products and those in clinical trials. Dextenza, INVELTYS, Dexycu and Bromsite are examples of DDS that have enjoyed success in clinical trials leading to FDA approval. Nanoparticles and ocular iontophoresis form the next wave of DDS that have the potential to replace topical steroids eye-drops as the treatment of choice for anterior segment inflammation. With the current relentless pace of ophthalmic drug delivery research, the pursuit of a new standard of treatment that eliminates the problems of low bioavailability and patient compliance may soon be realised.

We performed full circumferential imaging of the Schlemm’s canal (SC) of two human eyes using a Fourier domain mode-lock laser (FDML) based 1.66-MHz SS-OCT prototype at 1060 nm. Eight volumes with overlapping margins were acquired around the limbal area with customized raster scanning patterns designed to fully cover the SC while minimizing motion artifacts. The SC was segmented from the volumes using a semi-automated active contour segmentation algorithm, whose mean dice similarity coefficient was 0.76 compared to the manual segmentation results. We also reconstructed three-dimensional (3D) renderings of the 360° SC by stitching the segmented SCs from the volumetric datasets. Quantitative metrics of the full circumferential SC were provided, including the mean and standard deviation (SD) of the cross-sectional area (CSA), the maximum CSA, the minimum and maximum SC opening width, and the number of collector channels (CC) stemming from the SC.

Valencia Hui Xian Foo,1 Sarah En Mei Tan,2 David Ziyou Chen,3 Shamira A Perera,1,4 Charumathi Sabayanagam,4,5 Eva Katie Fenwick,4,5 Tina T Wong,1,4,5 Ecosse L Lamoureux4–6 1Singapore National Eye Centre, Singapore; 2Tan Tock Seng Hospital, Singapore; 3Department of Ophthalmology, National University of Singapore and National University Health System, Singapore; 4Singapore Eye Research Institute, Singapore National Eye Centre, Singapore; 5Centre for Quantitative Medicine, Duke-NUS Medical School, Singapore; 6Centre for Eye Research Australia, University of Melbourne, Melbourne, VIC, Australia Objective: The aim of this study was to evaluate patients’ dissatisfaction with overall and specific aspects of a tertiary glaucoma service and to determine their independent factors, including intraocular pressure (IOP) and visual acuity (VA). Methods: Patients, aged ≥21 years, from a specialist glaucoma service in a tertiary eye hospital in Singapore for at least 6 months, were recruited for this cross-sectional study between March and June 2014. All consenting patients completed a 7-area glaucoma-specific satisfaction questionnaire and one item related to satisfaction with overall glaucoma care. We determined the top three areas of dissatisfaction and overall dissatisfaction with the glaucoma service. We also explored the independent factors associated with overall and specific areas of patients’ dissatisfaction with their glaucoma care, including VA and IOP by using logistic regression models. Results: Of the 518 patients recruited, 438 (84.6%) patients completed the study. Patients’ dissatisfaction with the overall glaucoma service was 7.5%. The three areas of glaucoma service with the highest dissatisfaction rates were as follows: 1) explanation of test results (24.8%); 2) explanation of glaucoma complications (23.7%); and 3) advice on managing glaucoma (23.5%). Patients who were dissatisfied with the overall service had a worse mean VA compared with satisfied patients (logarithm of the minimum angle of resolution =0.41±0.43 vs 0.27±0.49, p=0.005), whereas mean IOP remained well-controlled in both the groups (13.55±2.46 mmHg vs 14.82±2.86 mmHg, p=0.014). In adjusted models, factors associated with overall dissatisfaction with glaucoma care included a pre-university education and above (odds ratio [OR] =8.06, 95% CI =1.57–41.27) and lower IOP (OR =0.83, 95% CI =0.71–0.98). Conclusion: Although less than one tenth of glaucoma patients were dissatisfied with the overall glaucoma service, one in four patients were dissatisfied with three specific aspects of care. A lower IOP, ironically, and education level were associated with overall dissatisfaction. Improving patients’ understanding of glaucoma test results, glaucoma complications, and disease management may increase patient satisfaction levels. Keywords: physician-patient relations, perception, attitude to health, surveys and questionnaires, quality of health care

Semra Ozdemir,1,* Hong King Wu,1,* Eric A Finkelstein,1 Tina T Wong2 1Signature Program in Health Services and Systems Research, Duke-NUS Medical School Singapore, 2Glaucoma Department, Singapore National Eye Centre, Singapore *These authors contributed equally to this work Aim: The objectives of this study were to explore parents' views about their children's use of regular eye drops and whether they would consider a sustained-release subconjunctival injection as a replacement for daily drops.Methods: A survey was conducted with 134 parents of children with chronic eye diseases at the Singapore National Eye Centre. Parents were asked their views about their children’s use of eye drops and were then presented with a discrete choice experiment that, via a series of trade-off tasks, allowed for estimating demand for a series of hypothetical subconjunctival injections that varied along product features, including interval between administrations, risk of complications, out-of-pocket cost and whether it is recommended by the patient’s treating physician.Results: Results showed that the vast majority of parents did not find administration of eye drops to be inconvenient (78%) nor did children complain about using daily eye drops (78%). Furthermore, only about half of parents whose child missed doses stated concerns about the consequences of non-compliance. The discrete choice experiment revealed that only one in fiveparents would consider a subconjunctival injection for their children. These parents tended to be more concerned about the consequences of non-compliance with eye drops, had children who administered the drops themselves or had other chronic disease requiring regular medication. Among these parents, risk of complications had the largest effect on injection uptake.Conclusion: This study shows that parents do not find administration of daily eye drops to be a significant burden. As a result, most would not consider a subconjunctival injection unless risk of complications was extremely small. Keywords: sustained-release subconjunctival injection, parent preferences, children’s use of eye drops

Purpose To evaluate the biomechanical properties (Young's modulus) of normal (control) and glaucomatous human iris using atomic force microscopy (AFM). Methods Iris tissue obtained from eighteen glaucomatous subjects (equal number of eyes with primary angle closure glaucoma (PACG) and primary open angle glaucoma (POAG) and five normal subjects who underwent elective eye surgery were subjected to the estimation of Young's modulus by AFM. Force measurements were done at room temperature using Nanowizard II BioAFM. The iris samples were immersed in the liquid media (PBS with 0.1% BSA) during force measurements. Young's modulus values were calculated for each recorded curve using JPK Data Processing Software, which uses a Hertz's contact model for spherical indenters fitted to the extend curves. Results The iris from the normal controls had the least Young's modulus (0.85 ± 0.31 kPa) while those from PACG patients had the highest Young's modulus (2.40 ± 0.82 kPa). The Young's modulus of PACG iris was significantly higher compared to that of the normal controls (P = 0.005) and POAG iris (P = 0.001). However, there was no significant difference in the Young's modulus of POAG iris (1.13 ± 0.36 kPa) compared to that of the normal controls (P = 0.511). Conclusions Variations in biomechanical properties of iris tissue may have a significant role in the pathogenesis of angle closure glaucoma. This study suggests the existence of fundamental biomechanical differences in eyes with angle closure versus open angle glaucoma. An understanding of this basis creates a new platform to understand disease pathology better and work on therapeutic strategies that will address the same.

Purpose The purpose of this study was to investigate whether the addition of primary angle closure glaucoma (PACG)–associated genetic loci allows improved detection of PACG, compared to anterior segment parameters measured by imaging. Design Case-control study. Methods Genotype data of the 8 PACG single-nucleotide polymorphisms (SNPs) (rs11024102 at PLEKHA7, rs3753841 at COL11A1, rs1015213 located between PCMTD1 and ST18 on Chromosome 8q, rs3816415 at EPDR1, rs1258267 at CHAT, rs736893 at GLIS3, rs7494379 at FERMT2, and rs3739821 mapping in between DPM2 and FAM102A) were available. Customized software was used to measure anterior segment optical coherence tomography (ASOCT) parameters, namely, anterior chamber depth, width, and area (ACD, ACW, and ACA) and lens vault (LV). Statistical analysis for positive predictive values was modeled using the receiver operating characteristic curve (AUC). Statistical significance comparing predictive power of the different parameters was calculated using permutation. Results A total of 388 PACG subjects and 751 controls with both ASOCT and genetic data were available for analysis. Anterior segment parameters including ACD, ACA, and LV had excellent predictive value (AUCs >0.94), except ACW (AUC=0.65), for identifying PACG. The inclusion of genetic risk alleles (either singly or as a composite genetic risk score for 8 genomewide association study SNPs) to ACD only provided a +0.50% improvement in reclassifying PACG cases and controls over and above the discriminatory value of ACD. This +0.50% improvement was not statistically significant (P > .05). Conclusions Although significant on their own, the incorporation of genetic data in the context of anterior segment imaging parameters like ACD provided only a marginal improvement of PACG detection.

Purpose Aquaporins (AQPs) facilitate transmembrane osmotic water transport and may play a role in iris fluid conductivity, which is implicated in the pathophysiology of glaucoma. In this study, we compared the iris expression of AQPs and aqueous osmolality between primary angle closure glaucoma (PACG), primary open-angle glaucoma (POAG), and nonglaucoma eyes. Methods AQP1-5 transcripts from a cohort of 36 PACG, 34 POAG and 26 nonglaucoma irises were measured by quantitative real-time PCR. Osmolality of aqueous humor from another cohort of 49 PACG, 50 POAG, and 50 nonglaucoma eyes were measured using an osmometer. The localization of AQP1 in both glaucoma and nonglaucoma irises was determined by immunofluorescent analysis. Results Of the five AQP genes evaluated, AQP1 and AQP2 transcripts were significantly upregulated in both PACG (3.48- and 8.07-fold, respectively) and POAG (3.12- and 11.58-fold, respectively) irises relative to nonglaucoma counterparts. The aqueous osmolalities of PACG (303.68 mmol/kg) and POAG (300.79 mmol/kg) eyes were significantly lower compared to nonglaucoma eyes (312.6 mmol/kg). There was no significant difference in expression of AQP transcripts or aqueous osmolality between PACG and POAG eyes. Conclusions PACG and POAG eyes featured significant increase in AQP1 and AQP2 expression in the iris and reduced aqueous osmolality compared to nonglaucoma eyes. These findings suggest that the iris may be involved in altered aqueous humor dynamics in glaucoma pathophysiology. Because PACG did not differ from POAG in both properties studied, it is likely that they are common to glaucoma disease in general.

Purpose To report the incidence and risk factors of elevated IOP following deep anterior lamellar keratoplasty (DALK). Design A retrospective case series. Methods A retrospective study investigating the 5-year incidence of raised IOP following DALK cases performed from 2004 to 2008 in a tertiary center. Patients with less than 6 months of follow-up were excluded. Elevated IOP was defined as IOP >21 mm Hg. Results An episode of elevated IOP occurred in 36.1% of cases (44/122 cases), 11.4% (n = 5) occurring within the first week. The average duration of raised IOP was 48.9 (SD: 65.5) days. Causes included pupil block from air, swollen grafts, and corticosteroid response. Surgical intervention to lower IOP was required in 3 cases (6.8%). In multivariate analyses, the use of Olopatadine 0.1% or cyclosporine eye drops before DALK (OR = 14.51, 95% CI = 1.43–147.23) and use of topical prednisolone acetate 1% compared with dexamethasone 0.1% post DALK (OR = 4.79, 95% CI = 0.73–31.52) were associated with higher rates of elevated IOP post DALK. At 5 years post DALK, 3 of 71 cases (4.48%) developed de novo glaucomatous field defects, and 1 case with pre-existing glaucoma had progression of glaucomatous field defect. Conclusions DALK was associated with a significant incidence of transiently elevated IOP postoperatively, but had a low incidence of de novo glaucoma at 5 years in our study. Risk factors for raised IOP post DALK included the use of topical prednisolone acetate 1% compared with dexamethasone 0.1%, and the use of Olopatadine 0.1% or any concentration of cyclosporine eye drops prior to DALK.

Purpose To investigate the patterns of visual field (VF) defects in primary angle-closure glaucoma (PACG) across different severity levels and to assess hemifield differences within each severity level. Design Cross-sectional study. Participants Three hundred four patients diagnosed with PACG were recruited from glaucoma clinics at a Singapore hospital. Methods Point-wise total deviation values were recorded from the static automated perimetry (Swedish interactive threshold algorithm standard program 24-2; Humphrey model 750 [Carl Zeiss Meditec, Dublin, CA]) printouts. Patients were excluded if they had unreliable VFs (fixation losses >33% and false-positive responses >15%), had undergone only 10-2 VF testing, had VF defects not typical of glaucoma, or had undergone cataract extraction. Mild, moderate, and severe VF loss were defined by a mean deviation of −6.00 dB or more, −6.01 to −12.00 dB, and −12.01 dB or less, respectively. Each hemifield was divided into regions according to glaucoma hemifield test sectors. The average mean deviation (MD) of each region was obtained using total deviation values. Main Outcome Measures Between- and within-hemifield differences of the regions across the severity levels. Results After excluding ineligible cases, 249 patients with PACG were included in the analysis. Mean age of the patients was 65.7±8.6 years, with a 1:1 gender ratio. The number of patients who had mild, moderate, and severe VFs was 72 (28.9%), 78 (31.3%), and 99 (39.8%), respectively. For between-hemifield comparisons, all regions in the superior hemifield had worse MDs compared with their counterparts in the inferior hemifield across the severity spectrum. Likewise, for within-hemifield comparisons, MDs of the regions gradually worsened with increasing distance from the fixation point. Conclusions In this group of clinic-based PACG patients, the superior hemifield was found to be affected more severely than the inferior hemifield, and the differences between them increased with worsening disease severity. The damage was consistently more pronounced in the nasal area.

Background This study aimed to evaluate differences in iris gene expression profiles between primary angle closure glaucoma (PACG) and primary open angle glaucoma (POAG) and their interaction with biometric characteristics. Design Prospective study. Participants Thirty-five subjects with PACG and thirty-three subjects with POAG who required trabeculectomy were enrolled at the Singapore National Eye Centre, Singapore. Methods Iris specimens, obtained by iridectomy, were analysed by real-time polymerase chain reaction for expression of type I collagen, vascular endothelial growth factor (VEGF)-A, -B and -C, as well as VEGF receptors (VEGFRs) 1 and 2. Anterior segment optical coherence tomography (ASOCT) imaging for biometric parameters, including anterior chamber depth (ACD), anterior chamber volume (ACV) and lens vault (LV), was also performed pre-operatively. Main Outcome Measures Relative mRNA levels between PACG and POAG irises, biometric measurements, discriminant analyses using genes and biometric parameters. Results COL1A1, VEGFB, VEGFC and VEGFR2 mRNA expression was higher in PACG compared to POAG irises. LV, ACD and ACV were significantly different between the two subgroups. Discriminant analyses based on gene expression, biometric parameters or a combination of both gene expression and biometrics (LV and ACV), correctly classified 94.1%, 85.3% and 94.1% of the original PACG and POAG cases, respectively. The discriminant function combining genes and biometrics demonstrated the highest accuracy in cross-validated classification of the two glaucoma subtypes. Conclusions Distinct iris gene expression supports the pathophysiological differences that exist between PACG and POAG. Biometric parameters can combine with iris gene expression to more accurately define PACG from POAG.

Purpose To determine the effectiveness of argon laser peripheral iridoplasty (ALPI) in primary angle closure (PAC) and primary angle-closure glaucoma (PACG). Design Randomized controlled trial. Participants Eighty PAC or PACG subjects who underwent laser iridotomy (LI) and had at least 180° of persistent appositional angle closure and intraocular pressure (IOP) of more than 21 mmHg were enrolled. Methods Subjects were randomized to receive either 360° ALPI (Visulas 532s; Carl Zeiss Meditec, Jena, Germany) or medical therapy (Travoprost 0.004%; Alcon-Couvreur, Puurs, Antwerp, Belgium). Repeat ALPI was performed if the IOP reduction was less than 20% from baseline along with inadequate angle widening at the month 1 or month 3 visit. Intraocular pressure was controlled with systematic addition of medications when required. Main Outcome Measures The primary outcome measure was success rates after ALPI at 1 year. Complete success was defined as an IOP of 21 mmHg or less without medication, and qualified success was defined as an IOP of 21 mmHg or less with medication. Failure was defined as an IOP more than 21 mmHg despite additional medications or requiring glaucoma surgery. Results Forty subjects (51 eyes) were randomized to ALPI and 40 subjects (55 eyes) were randomized to medical therapy. Complete success (IOP ≤21 mmHg without medication) was achieved in 35.0% eyes of the ALPI group compared with 85.0% of eyes in the prostaglandin analog (PGA) group ( P P = 0.003). The IOP decreased by 4.9 mmHg (95% confidence interval [CI], 3.5–6.3 mmHg) in the ALPI group ( P P P = 0.01). No treatment-related complications were recorded in either group. Conclusions After 1 year, ALPI was associated with higher failure rates and lower IOP reduction compared with PGA therapy in eyes with persistent appositional angle closure and raised IOP after LI.

Glaucoma filtration surgery (GFS) commonly fails due to excessive fibrosis. As collagen structure aberrations is implicated in adverse fibrotic progression, this study aims to uncover collagen organization alterations during postoperative scarring. Via quantitative second harmonic generation/ two photon excitation multiphoton imaging, we reveal the scar development and phenotype in the mouse model of conjunctival scarring. We also show that multiphoton imaging corroborated the collagen ultrastructure anomaly characteristic of the SPARC-/- mouse postoperative conjunctiva. These data improve our understanding of postoperative conjunctival scarring and further enhance the utility of this model for the development of anti-fibrotic therapeutics for GFS.

Objectives: Clinical translation of siRNA therapeutics has been severely limited due to the lack of stable and sustained siRNA delivery systems. Furthermore, when nanocarrier systems with siRNA are administered systemically to treat diseases, insufficient doses reach the target tissue. Here we report the successful development of a new nanocarrier system for the management of fibrosis. Methods: The new carrier has a hydroxyapatite core, with alternating layers of siRNA and a cationic peptide. The siRNA used here targets secreted protein acidic and rich in cysteine (SPARC), a key matricellular protein involved in the regulation of collagen fibrillogenesis and assembly. We have also used FRET studies to elucidate the fate of the particles inside cells, including the mechanistic details of layer-by-layer detachment. Results: In vitro studies using murine conjunctiva fibroblasts show sustained release over 2 weeks, and that such released siRNA sustained SPARC knockdown without affecting cell growth, and maintained siRNA presence in the cells for at least two weeks with a single-dose treatment. Release studies of siRNA from particles in vitro gave insight on how the particles delivered prolonged gene-silencing effects. Conclusion: A single treatment of the layer-by-layer nanoparticle designed can achieve sustained gene silencing over 2 weeks. Localized delivery of stabilized siRNA with sustained-release capabilities opens the door for many other applications of siRNA-based gene regulation. NRF (Natl Research Foundation, S’pore) NMRC (Natl Medical Research Council, S’pore) MOH (Min. of Health, S’pore)

We aim to evaluate changes in the ocular surface in a cohort of post-trabeculectomy patients and whether these were associated with conjunctival inflammatory gene expression.This is a single-arm interventional cohort performed in a tertiary referral center. These were assessed: dry eye symptom questionnaire, tear osmolarity, Schirmer's test, non-invasive tear break up time (BUT), conjunctival redness and corneal fluorescein staining evaluation. Conjunctival impressions were performed using Eyeprim, and after RNA extraction, transcripts of 255 inflammatory genes were analysed using the Nanostring nCounter assay.Thirty three patients were recruited with age 66.88 ± 9.76 at baseline, with a predominance of men. There was a significant decrease in inferior corneal staining at 6 months (p 0.05) (n = 22) and significant decrease in tear osmolarity at 12 months (p 0.01) (n = 27). No patient required glaucoma eyedrops post-surgery up to 3 years. At baseline 31/33 transcript profiles passed the quality control, and after normalization, 249 transcripts were subsequently analysed. Increased discomfort was associated with higher Protein Tyrosine Kinase-2 at the cross-sectional analysis at baseline. Lower baseline complement factor-D and higher levels of Mitogen associated kinase-8, MAP3K1 and MyD88, were associated with presence of corneal staining at 6 months. Nine genes, including the proinflammatory lipo-oxygenase (ALOX5) showed a significantly reduced level at 3 years (n = 5).Glaucoma surgery may confer long term beneficial effect on the ocular surface, if anti-glaucoma eyedrops are no longer necessary. This may be due to reduced expression of conjunctival proinflammatory genes and immune-related genes.