Extracellular vesicle encapsulated nicotinamide delivered via a trans-scleral route provides retinal ganglion cell neuroprotection.

The progressive and irreversible degeneration of retinal ganglion cells (RGCs) and their axons is the major characteristic of glaucoma, a leading cause of irreversible blindness worldwide. Nicotinamide adenine dinucleotide (NAD) is a cofactor and metabolite of redox reaction critical for neuronal survival. Supplementation with nicotinamide (NAM), a precursor of NAD, can confer neuroprotective effects against glaucomatous damage caused by an age-related decline of NAD or mitochondrial dysfunction, reflecting the high metabolic activity of RGCs. However, oral supplementation of drug is relatively less efficient in terms of transmissibility to RGCs compared to direct delivery methods such as intraocular injection or delivery using subconjunctival depots. Neither method is ideal, given the risks of infection and subconjunctival scarring without novel techniques. By contrast, extracellular vesicles (EVs) have advantages as a drug delivery system with low immunogeneity and tissue interactions. We have evaluated the EV delivery of NAM as an RGC protective agent using a quantitative assessment of dendritic integrity using DiOlistics, which is confirmed to be a more sensitive measure of neuronal health in our mouse glaucoma model than the evaluation of somatic loss via the immunostaining method. NAM or NAM-loaded EVs showed a significant neuroprotective effect in the mouse retinal explant model. Furthermore, NAM-loaded EVs can penetrate the sclera once deployed in the subconjunctival space. These results confirm the feasibility of using subconjunctival injection of EVs to deliver NAM to intraocular targets.

Five-Year Outcomes of Single Trabecular Microbypass Stent (iStent®) Implantation with Phacoemulsification in Korean Patients.

INTRODUCTION: This study reports the long-term intraocular pressure (IOP)-lowering efficacy and safety of a single trabecular microbypass stent (iStent®; Glaukos Corp., San Clemente, CA, USA) for medically controlled open-angle glaucoma in Korean patients. METHODS: This retrospective observational study included 42 eyes of 35 patients with primary open-angle glaucoma (POAG). All subjects underwent single first-generation iStent® implantation with phacoemulsification by a single surgeon with 5 years follow-up. The primary outcomes were changes in IOP and the number of antiglaucoma medications compared to the preoperative values. The secondary outcome was the proportion of eyes with IOP ≤ 18 mmHg without medication, ≤ 15 mmHg without medication, and ≤ 18 mmHg with or without medication. Adverse events and need for secondary glaucoma surgery were also recorded. RESULTS: The mean IOP decreased from 15.8 ± 2.8 to 13.8 ± 1.7 mmHg and the mean number of medications was reduced from 2.24 ± 1.18 to 0.83 ± 1.12, respectively, at year 5. At 3 and 5 years, 80.6% and 78.6% of eyes, respectively, were receiving fewer medications than preoperative numbers. In contrast, only 50% of eyes on four preoperative medications showed medication reductions at 5 years. At years 3 and 5, 61.3% and 53.5% of eyes achieved IOP ≤ 18 mmHg without medication, whereas 90.3% and 89.3% of eyes achieved ≤ 18 mmHg regardless of medication use, respectively. Four eyes required additional glaucoma surgery (two Ahmed glaucoma valve implantations, one trabeculectomy, and one XEN 45 Gel Stent implantation), and all were receiving four preoperative antiglaucoma medications. Transient IOP elevation (14.3%) was the most common complication, followed by five hyphema, one stent obstruction, one stent malposition, and one severe anterior chamber reaction. CONCLUSION: This study demonstrated a good safety profile and sustained IOP reduction after the implantation of a single trabecular microbypass stent (iStent®) with phacoemulsification in Korean patients. The majority of subjects with POAG showed a relatively good response; however, eyes receiving a higher number of medications preoperatively (especially four medications) had difficulty achieving a low target IOP.

XEN Gel Stent for Conjunctiva with Minimal Mobility Caused by Scleral Encircling: A Case Report.

This case report describes the successful use of a XEN gel stent for controlling intraocular pressure (IOP) in a patient who had previously undergone scleral encircling for rhegmatogenous retinal detachment. The patient had very limited mobile conjunctiva due to scarring caused by the earlier surgery, which limited their options for glaucoma surgery. The XEN gel stent, a minimally invasive glaucoma surgery (MIGS) procedure that does not require opening the conjunctiva, was implanted in the subconjunctival space using an ab interno approach. Postoperative blebs were imaged using anterior segment optical coherence tomography, and IOP was monitored over six months. This study found that the XEN gel stent effectively controlled the IOP, and there were no complications during or after surgery. This case report may expand the indication for the XEN gel stent, which could be considered a viable option for patients who have undergone scleral buckling and have limited mobile conjunctiva.

Comparisons of efficacy and safety between preserved and preservative-free brimonidine tartrate in glaucoma and ocular hypertension: a parallel-grouped, randomized trial.

This multicenter (four institutions), randomized, investigator-masked, parallel-group clinical trial evaluated and compared the efficacy and safety of preservative-free and preserved brimonidine tartrate 0.15% in open-angle glaucoma and ocular hypertension. Sixty eyes of 60 patients with intraocular pressure (IOP) ≥ 15 mmHg diagnosed with open-angle glaucoma or ocular hypertension were randomized to preserved (n = 31) and preservative-free (n = 29) brimonidine groups. The enrolled eyes received brimonidine monotherapy three times daily. Main outcome measures were corneal/conjunctival staining score, ocular surface disease index, patient satisfaction score, drug tolerance, and drug adherence rate 12 weeks post first administration. Secondary outcome measurements included visual acuity, IOP, drug tolerance, tear-film break-up time, hemodynamic changes including blood pressure and heart rates, and ocular adverse events. After 12 weeks, both preserved and preservative-free groups showed similar IOP reduction, corneal and conjunctival staining scores, drug tolerance, and adherence rates. The preservative-free group showed significantly better tear-film break-up time and higher patient satisfaction regarding drug use and management. Systolic and diastolic blood pressure reductions during the 12 weeks were significantly lower in the preserved group than in the preservative-free group. Preservative-free brimonidine tartrate showed comparable efficacy and safety, better corneal tear film stability, and patient satisfaction than preserved brimonidine.

Bleb Analysis Using Anterior Segment Optical Coherence Tomography and Surgical Predictors of XEN Gel Stent.

PURPOSE: To determine the early predictors of surgical success 6 months after XEN surgery among clinical parameters, including anterior segment optical coherence tomography (AS-OCT). METHODS: A total of 31 eyes with medically uncontrolled primary open-angle glaucoma or pseudoexfoliation glaucoma was enrolled retrospectively. Using AS-OCT, XEN tip location was categorized into intraconjunctival, intratenon, or uviform at day 1 and blebs were classified into no or low, high sparse, high thick, cystic, or mixed walls at month 6. Using slit-lamp photography, blebs were classified into no or low, localized avascular, diffuse avascular, localized vascular, or diffuse vascular blebs at month 6. Surgical success was defined as an intraocular pressure (IOP) of less than 14 mm Hg. RESULTS: Intraconjunctival and intratenon locations of the tip mostly created a high sparse wall, whereas the uviform type mostly created no or low wall and no or low bleb. The uviform type was linked to pseudoexfoliation glaucoma. A high sparse wall and diffuse avascular bleb showed a lower mean IOP than a high thick wall and localized vascular bleb. In the multivariate analysis, female sex and IOP at week 1 were early predictors of surgical success (8.45 times and 33.1% per 1 mm Hg-decrease, respectively). CONCLUSIONS: Bleb evaluation using AS-OCT is valuable to correlate tip location and bleb morphology with clinical profiles, considering that a lower early postoperative IOP is linked to surgical success. TRANSLATIONAL RELEVANCE: Bleb analysis using AS-OCT on day 1 could help to predict bleb morphology after 6 months, which is important to maintain the functioning bleb in the longer term.

Efficacy and safety of newly developed preservative-free latanoprost 0.005% eye drops versus preserved latanoprost 0.005% in open angle glaucoma and ocular hypertension: 12-week results of a randomized, multicenter, controlled phase III trial.

AIM: To evaluate the therapeutic efficacy, safety and tolerability of newly developed preservative-free (PF) latanoprost generic [TJO-002] and compare it with benzalkonium chloride (BAK)-preserved latanoprost [Xalatan®] in patients with primary open angle glaucoma (POAG) and ocular hypertension (OHT). METHODS: Included patients were aged ≥19y with POAG/OHT. After a washout period, patients with IOP 21-35 mm Hg at 9 a.m. were enrolled. After a full ophthalmic and glaucoma examination, 144 patients with POAG and OHT participated in this study. Subjects were randomly assigned either PF latanoprost (74 eyes) or BAK-preserved latanoprost (70 eyes). All subjects were examined at 4, 8, and 12wk after first administration. At each follow-up visit, IOP was measured at 9 a.m. and 5 p.m. and compliance was assessed. Throughout the study, all adverse events were recorded and monitored by the masked investigators who measured IOP. RESULTS: Both groups showed a statistically significant decrease of average diurnal IOP at 12wk compared to baseline (-7.21±3.10 mm Hg in the PF latanoprost group and -7.02±3.17 mm Hg in the BAK latanoprost group, both P<0.0001). There was no statistically significant diurnal IOP variation between the groups. In terms of tolerability, pruritus, burning/stinging, and sticky eye sensation, severity was significantly lower in the PF latanoprost group than in the BAK latanoprost group (P<0.05). CONCLUSION: PF latanoprost has at least similar efficacy in terms of IOP reduction and better tolerability compared with BAK latanoprost.

Clinical Outcomes After Second-Generation Trabecular Microbypass Stents (iStent inject®) with Phacoemulsification in Korean Patients.

INTRODUCTION: To evaluate the intraocular pressure (IOP)-lowering effect of second-generation trabecular microbypass stents (iStent® inject) with cataract extraction (combination group) and compare refractive changes in the combination group and the control (phacoemulsification only) group. METHODS: This retrospective case-control study included 36 eyes with cataract and medically controlled open-angle glaucoma with IOP < 21 mmHg and 100 nonglaucomatous eyes with cataract. Data were collected preoperatively and for 6 months postoperatively. Data included IOP, number of glaucoma medications, corrected distance visual acuity, and mean absolute error (MAE) from target refraction, and astigmatic vector analysis. Surgical success for the combination group was defined according to three criteria: (A) IOP < 15 mmHg without medication, (B) IOP < 18 mmHg without medication, and (C) IOP < 18 mmHg with or without medication. RESULTS: In the combination group, mean IOP was reduced from 15.1 ± 2.9 mmHg to 12.5 ± 2.0 mmHg, and the mean number of medications decreased from 1.9 ± 1.0 to 0.4 ± 0.8 at postoperative 6 months (both P < 0.001). Surgical success rates were 77.8%, 83.3%, and 97.2% at 6 months by criteria A, B, and C, respectively. Mean IOP was reduced from 14.3 ± 2.7 mmHg to 13.1 ± 2.1 mmHg at 1 month in the control group (P < 0.001). The MAE was 0.33 ± 0.26 D, and 83.3% of eyes had spherical equivalent difference within 0.50 D in the combination group (0.38 ± 0.33 D and 76.0% in the control group; P = 0.309 and P = 0.363, respectively). Preoperative and postoperative centroid values were 0.51 D @ 1° and 0.66 D @ 178°, respectively (0.23 D @ 176° and 0.66 D @ 1° in the control group). There were no statistical differences between the two groups with respect to preoperative and postoperative mean absolute values (P = 0.154 and P = 0.322, respectively). CONCLUSIONS: On the basis of our results using Korean real-world interim experience, iStent® inject with cataract extraction has favorable IOP-lowering effects and minimally impacts refractive outcomes.

Risk factors for periorbital dermatitis in patients using dorzolamide/timolol eye drops.

This study assessed the clinical risk factors for periorbital dermatitis (PD) after using dorzolamide/timolol eye drops in a total of 1282 glaucoma patients. Both the PD(+) group and the PD(-) group were evaluated using clinical data such as age, sex, dosing duration, presence of benzalkonium chloride (BAK) in the formulation, ocular surgery history (e.g. cataract or glaucoma operations), height, weight, personal history of systemic hypertension, smoking, alcohol consumption, intraocular pressure, best-corrected visual acuity (BCVA), central corneal thickness, axial length, and visual field index (VFI). Univariate analyses showed that shorter dosing duration, higher rate of BAK-included cases, worse BCVA, worse VFI, more systemic hypertension history, and more ocular surgery history were more associated with the PD(+) group than the PD(-) group. The BAK(-) group showed a lower PD rate than the BAK-included group, which was supported by the Kaplan-Meier analysis (log-rank test, p = 0.0014). Multivariate analyses revealed that the probability of PD increased by 8 times if they had a history of ocular surgery and increased by 2.3% when the VFI decreased by 1% (Cox's hazard regression test, p < 0.001). Therefore, a preservative-free dorzolamide/timolol can benefit the subjects for those who had ocular surgery or who have worse VFI.

Combined argon laser peripheral iridoplasty and Nd: YAG laser shock wave therapy for recurrent XEN gel stent obstruction due to iris incarceration: A case report.

INTRODUCTION: Despite its proven effectiveness and safety profile, the XEN Gel Stent (Allergan Inc., CA, USA) has a small lumen and is therefore likely to become occluded by fibrin, a blood clot, or even the iris. However, few studies have investigated XEN-iris occlusion and how to manage this condition. We describe the first case report of recurrent XEN gel stent obstruction by iris incarceration, which was resolved following a combined treatment with argon laser peripheral iridoplasty (ALPI) and low-energy neodymium-doped yttrium aluminum garnet (Nd: YAG) laser shock wave treatment. PATIENT INFORMATION: A 74-year-old Korean male underwent uncomplicated XEN gel stent implantation and presented with low intraocular pressure (IOP) with a well-functioning filtering bleb during the first postoperative week. On postoperative day 10, the XEN lumen was occluded by the iris and demonstrated an IOP spike of 33 mmHg. Despite the use of pilocarpine, the iris incarceration persisted. Therefore, surgery to reposition the XEN stent was attempted using a gonio-prism and intraocular forceps. After the first revision surgery, the IOP and stent position were stable for 2 weeks. However, recurrent partial obstruction of the stent by the iris, pigment dispersion into the intraluminal space, and an elevated IOP of 24 mmHg were observed later. DIAGNOSIS: Recurrent XEN gel stent occlusion by the iris and intraluminal pigment dispersion. INTERVENTIONS: Combined ALPI and low energy Nd: YAG laser shock wave therapy. OUTCOMES: IOP dropped from 24 mmHg to 10 mmHg immediately and continued to be well-controlled until 3 months later (range: 8-12 mmHg). CONCLUSIONS: To the best of our knowledge, this is the first case report of the efficacy of combined laser treatment for relieving recurrent XEN implant occlusion by the iris. This combination laser treatment might be a relatively safe rescue treatment to restore the patency of a XEN gel stent occluded by the iris, even in cases with recurrent XEN stent obstruction after surgical repositioning.

Comparison of ocular surface assessment and adherence between preserved and preservative-free latanoprost in glaucoma: a parallel-grouped randomized trial.

Given that nonadherence is related to subject characteristics and drug tolerance and preserved eye drops tend to be more intolerable than preservative-free ones, we conducted a phase 4, parallel-grouped, investigator-blind, active-control, randomized, multicenter study. A total of 51 patients with intraocular pressure (IOP) ≥ 15 mmHg diagnosed with open-angle glaucoma or ocular hypertension were randomly assigned to the preserved latanoprost group (n = 26) and the preservative-free latanoprost group (n = 25). The efficacy variables were corneal/conjunctival staining grade, Ocular Surface Disease Index (OSDI), adherence at 12 weeks after the first administration; corneal/conjunctival staining grade at 4 weeks; and IOP, tear break-up time (TBUT), and hyperemia score at 4 and 12 weeks. The safety variables included visual acuity and drug tolerance questionnaire results. There was no statistically significant difference in corneal/conjunctival staining grade, OSDI, or TBUT between the groups at 4 and 12 weeks. However, the adherence rate was higher and the hyperemia score was lower in the preservative-free group than in the preserved group. The severity and duration of stinging/burning sensation were lower in the preservative-free group than in the preserved group. Overall, preservative-free latanoprost showed better ocular tolerance assessed by hyperemia scores and stinging/burning symptoms following higher adherence than preserved latanoprost.

Comparison of the Safety and Efficacy between Preserved and Preservative-Free Latanoprost and Preservative-Free Tafluprost.

In this study, we investigated the effect of preservative-free (PF) 0.0015% tafluprost (TA), to the preservative containing (PC) and the PF 0.005% latanoprost (LA) in Korean subjects. This study was conducted as a multi-center, randomized, investigator-blind, active controlled, parallel-group, clinical trial in adult patients (≥19 years) with open-angle glaucoma (OAG) and ocular hypertension (OHT). After a washout period, patients with an IOP between 15 and 35 mmHg were enrolled and evaluated the efficacy, safety, and compliance at 4, 8 and 12 weeks after the first administration. A total of 137 OAG and OHT patients were randomized. Statistically significant reductions in IOP were observed in all groups. Twelve weeks after each eye drop instillation, the mean IOP reduction was -4.59 ± 2.70 mmHg (-24.57 ± 13.49%) in the PC-LA group, -4.52 ± 2.17 mmHg (-24.41 ± 11.38%) in the PF-LA, and -3.14 ± 2.83 mmHg (-17.22 ± 14.57%) in the PF-TA group. The PF-LA showed significantly better responsiveness than did PF-TA. PF-LA was better tolerated than was PC-LA. There were no adverse events that led to cessation of eye drop use in any of the groups. In conclusion, IOP decreased similarly across the groups. PF-LA may provide a good choice for OAG patients with ocular surface diseases.

Nicotinamide provides neuroprotection in glaucoma by protecting against mitochondrial and metabolic dysfunction.

Nicotinamide adenine dinucleotide (NAD) is a REDOX cofactor and metabolite essential for neuronal survival. Glaucoma is a common neurodegenerative disease in which neuronal levels of NAD decline. We assess the effects of nicotinamide (a precursor to NAD) on retinal ganglion cells (the affected neuron in glaucoma) in normal physiological conditions and across a range of glaucoma relevant insults including mitochondrial stress and axon degenerative insults. We demonstrate retinal ganglion cell somal, axonal, and dendritic neuroprotection by nicotinamide in rodent models which represent isolated ocular hypertensive, axon degenerative, and mitochondrial degenerative insults. We performed metabolomics enriched for small molecular weight metabolites for the retina, optic nerve, and superior colliculus which demonstrates that ocular hypertension induces widespread metabolic disruption, including consistent changes to α-ketoglutaric acid, creatine/creatinine, homocysteine, and glycerophosphocholine. This metabolic disruption is prevented by nicotinamide. Nicotinamide provides further neuroprotective effects by increasing oxidative phosphorylation, buffering and preventing metabolic stress, and increasing mitochondrial size and motility whilst simultaneously dampening action potential firing frequency. These data support continued determination of the utility of long-term nicotinamide treatment as a neuroprotective therapy for human glaucoma.

Clinical Efficacy of Custom-built Software for the Early Detection of Glaucoma: A Comparison of Axial-length and Major Retinal Artery Location Data.

PURPOSE: To assess the clinical efficacy for early detection of glaucoma using custom-built image software visualizing translucent retinal nerve fiber layer thickness (RNFLT) that is graphed based on a normative database. METHODS: This prospective study was conducted using a normative database constructed with RNFLT data of 151 healthy Korean eyes. The reference lines of the mean, the lower 5%, and the lower 1% limit were visualized as a translucent RNFLT graph produced by our software after inputting each subject's major retinal artery position and overlaying the results onto the RNFLT measurements. Fifty-eight additional healthy control and 79 early-glaucoma eyes were collected for the validation group. If a subject's RNFLT graph was outside the reference line of the lower 1% limit, the graph was defined as abnormal. The lower 1% limit, which was generated by three criteria (criterion 1, built-in software; criterion 2, axial-length data; criterion 3, major retinal artery data), was used to address the difference of agreement with a standard answer. RESULTS: For criteria 1, 2, and 3, the accuracy of our custom-built software was significantly higher than that of the manufacturer's database (kappa of 0.475 vs. 0.852 vs. 0.940; sensitivity of 62.0% vs. 91.1% vs. 97.5%, respectively) maintaining high specificity (87.9% vs. 94.8% vs. 96.6%, respectively). CONCLUSIONS: The custom-built imaging software with the constructed RNFLT normative database showed high clinical efficiency for early detection of glaucoma with negligible user-related variability.

Clinical and Biological Evaluations of Biodegradable Collagen Matrices for Glaucoma Drainage Device Implantation.

Purpose: To characterize the clinical and biological properties of biodegradable collagen matrices (BCMs) for possible glaucoma drainage device implantation. Methods: A total of 68 refractory glaucoma eyes, followed up postoperatively for at least 6 months, were consecutively enrolled after retrospective chart review. The BCM-augmented Ahmed valve implantations (BAAVI) using our Ologen-6 and Ologen-7 valves were performed and compared with a conventional method. Complete surgical success was defined as an IOP of ≤21 mm Hg (IOP 1) or ≤17 mm Hg (IOP 2) without antiglaucoma medications. Qualified success was defined as an IOP ≤21 mm Hg with or without antiglaucoma medications. The biological properties of each BCM were assessed by enzymatic degradation rates via collagenase under ocular physiological conditions. Results: The mean ages and preoperative IOPs were similar for the groups. In the conventional, BAAVI with Ologen-6, and BAAVI with Ologen-7 groups, complete success rates with target IOP 1 were 29.2%, 40.0%, and 66.7%; those with target IOP 2 were 12.5%, 30.0%, and 45.8%; qualified success rates were 45.8%, 55.0%, and 75.0%, respectively. The enzymatic degradation rate of Ologen-7 was significantly slower than that of Ologen-6 (12.5 × 10-3 vs. 28.8 × 10-3). Conclusions: The surgical success rate was highest in the Ologen-7 BAAVI group, with the lowest dependency on postoperative antiglaucoma medication use compared with the conventional and Ologen-6 BAAVI groups. The clinical results correlated with the different biological and physicochemical properties based on the degree of enzymatic degradation and on the structural morphology.

Agreement of New Automated Matched Alternation Flicker using Undilated Fundus Photography for the Detection of Glaucomatous Structural Change.

PURPOSE: To determine the agreement among glaucoma experts and general ophthalmologists regarding detection of glaucomatous structural changes using a new automated matched alternation flicker (AMAF) method with fundus photographs (FPs) of undilated eyes. METHODS: Sixty-six pairs of FPs of normal tension glaucoma patients were collected. FPs were taken at intervals of more than 12 months. Alternating flicker images were created using a new AMAF application. In a blinded manner, two glaucoma experts and two general ophthalmologists compared the presence of glaucomatous structural changes using either the AMAF method or the side-by-side comparison method. The interobserver and intraobserver agreements were compared using the Bland and Altman plot analysis. RESULTS: The glaucoma experts detected more glaucoma progression using the AMAF method (average, 50.7%) compared with the side-by-side method (average, 32.5%). General ophthalmologists detected more glaucomatous progression with the AMAF method (average, 40.9%) than with the side-by-side method (average, 25.0%). The AMAF method showed fair to substantial interreader agreement (k = 0.511-0.724) and fair to perfect intrareader agreement (k = 0.631-0.943). Interreader and intrareader agreements using the AMAF method were better for the glaucoma experts compared with the general ophthalmologists. CONCLUSIONS: The AMAF method showed more changes in FPs than the classical side-by-side comparison method. Regarding inter- and intrareader agreements, agreement for the glaucoma experts was best using the AMAF method, but for the general ophthalmologists agreement was best using the side-by-side comparison method.

Protective effect of etanercept, an inhibitor of tumor necrosis factor-α, in a rat model of retinal ischemia.

BACKGROUND: To assess the neuroprotective effect of etanercept (Enbrel®) which is a commercialized Tumor necrosis factor-α (TNF-α) inhibitor on axonal injury in an animal model of acute ischemia. METHODS: Acute ischemia was induced by intraocular pressure elevation in 36 rats. The treatment groups underwent subcutaneous injection of etanercept (0.3 or 1.0 mg/kg) three times per week up to 4 weeks. The control groups were treated in the same manner using the same volume of phosphate-buffered saline (PBS). Optic nerve damage was evaluated by counting the number of axons under a transmission electron microscope. Microglial cell activity was assessed using Iba1 and CD68. RESULTS: After induction of ischemia, the ratio of preserved axons was significantly greater in the 2-week 1.0-mg/kg etanercept-treated group than in the PBS-treated group (p = 0.062). The 4-week 0.3-mg/kg and 1.0-mg/kg etanercept-treated groups also showed significantly higher ratios of preserved axons than did the PBS-treated group (p = 0.021 and 0.003, respectively). The expression of Iba1 and CD68 in the optic nerve was lower in the etanercept-treated groups than in the PBS-treated groups. Immunohistochemical staining using rabbit anti-Iba1 antibody showed that the amount of microglia at the optic nerve head was noticeably lower in the etanercept-treated groups than in the PBS-treated groups. CONCLUSIONS: Etanercept significantly suppressed optic nerve injury in this rat model of acute ischemia. This in vivo study suggests that etanercept might be a novel neuroprotective treatment agent for TNF-α-related disease.

Human umbilical cord blood mononuclear cells and chorionic plate-derived mesenchymal stem cells promote axon survival in a rat model of optic nerve crush injury.

The use of mesenchymal stem cells (MSCs) in cell therapy in regenerative medicine has great potential, particularly in the treatment of nerve injury. Umbilical cord blood (UCB) reportedly contains stem cells, which have been widely used as a hematopoietic source and may have therapeutic potential for neurological impairment. Although ongoing research is dedicated to the management of traumatic optic nerve injury using various measures, novel therapeutic strategies based on the complex underlying mechanisms responsible for optic nerve injury, such as inflammation and/or ischemia, are required. In the present study, a rat model of optic nerve crush (ONC) injury was established in order to examine the effects of transplanting human chorionic plate-derived MSCs (CP­MSCs) isolated from the placenta, as well as human UCB mononuclear cells (CB-MNCs) on compressed rat optic nerves. Expression markers for inflammation, apoptosis, and optic nerve regeneration were analyzed, as well as the axon survival rate by direct counting. Increased axon survival rates were observed following the injection of CB­MNCs at at 1 week post-transplantation compared with the controls. The levels of growth-associated protein-43 (GAP­43) were increased after the injection of CB­MNCs or CP­MSCs compared with the controls, and the expression levels of hypoxia-inducible factor-1α (HIF-1α) were also significantly increased following the injection of CB-MNCs or CP-MSCs. ERM-like protein (ERMN) and SLIT-ROBO Rho GTPase activating protein 2 (SRGAP2) were found to be expressed in the optic nerves of the CP­MSC-injected rats with ONC injury. The findings of our study suggest that the administration of CB­MNCs or CP­MSCs may promote axon survival through systemic concomitant mechanisms involving GAP­43 and HIF­1α. Taken together, these findings provide further understanding of the mechanisms repsonsible for optic nerve injury and may aid in the development of novel cell-based therapeutic strategies with future applications in regenerative medicine, particularly in the management of optic nerve disorders.

Semi-automated counting method of axons in transmission electron microscopic images.

OBJECTIVE: To evaluate the accuracy of a new semi-automated method for counting axons in transmission electron microscopic (TEM) images. PROCEDURES: Optic nerve cross sections were obtained from both eyes of Sprague Dawley rats after unilateral induction of chronic ocular hypertension. TEM images (3000× magnification) of cross sections were evaluated by both semi-automated and manual counting methods. The semi-automated counting method was performed using ImageJ software after simple image optimization, and the resulting estimate of axon damage was compared with semiquantitative damage grading scale from light microscopic (LM) images. RESULTS: Axon counts obtained from the semi-automated method were strongly correlated with those obtained from the manual counting method (Pearson's correlation coefficient r = 0.996, P < 0.001) and from the full manual count from LM images (Spearman's ρ = 0.973, P < 0.001). The semi-automated method measured axonal damage with an error of 0.94 ± 3.16% (mean ± standard deviation), with worse axonal damage associated with greater error. Interobserver and intra-observer variability in axons counts were low (Spearman's ρ = 0.999, P < 0.005). The results of the semi-automated counting method were highly correlated with semiquantitative damage grading scale (Spearman's ρ = 0.965, P < 0.001). CONCLUSION: Results of our semi-automated method for counting axons in TEM images were strongly correlated with those of conventional counting methods and showed excellent reproducibility.

Bleb Analysis and Short-Term Results of Biodegradable Collagen Matrix-Augmented Ahmed Glaucoma Valve Implantation: 6-Month Follow-up.

PURPOSE: To evaluate the short-term efficacy of a biodegradable collagen matrix (BCM) as an adjuvant for Ahmed valve implantation surgery to prevent the hypertensive phase. METHODS: This prospective study included 43 refractory glaucoma eyes, all followed for 6 months. Refractory glaucoma was defined as an IOP higher than 20 mm Hg with antiglaucoma eye drops without previous glaucoma surgery. Conventional method was performed in 21 eyes and BCM-augmented Ahmed valve implantation (BAAVI) in 22 eyes. In the BAAVI group, a 10 × 10 × 2-mm BCM was sutured on an Ahmed glaucoma valve FP7 model. Complete success was defined as an IOP of 21 mm Hg or lower (target IOP 1) or 17 mm Hg or lower (target IOP 2) without antiglaucoma medications and qualified success as an IOP of 21 mm Hg or lower with or without medications. Maximal bleb thickness was measured using anterior segment optical coherence tomography images. RESULTS: The preoperative IOPs and numbers of preoperative antiglaucoma medications were similar for both groups. Complete target IOP 1 success rates were 38.1% and 86.4%, complete target IOP 2 success rates were 19.0% and 59.1%, and qualified success rates were 52.4% and 90.9% in the conventional and BAAVI groups, respectively (P < 0.05). The hypertensive phase rate was lower in the BAAVI group (4.5% vs. 47.6%, P = 0.002). Maximal bleb thickness was increased in the BAAVI group on postoperative days 30 and 180 (P < 0.05). CONCLUSIONS: Success rates were higher in the BAAVI group than in the conventional group with the change of bleb morphology. Furthermore, use of BCM significantly decreased the need for antiglaucoma medications for at least 6 months postoperatively.