Effects of Antiglaucoma Drugs on Calcium Mobility in Cultured Corneal Endothelial Cells

The aim of this study was to estimate the effects of various antiglaucoma drugs including betaxolol, timolol, levobunolol, brimonidine, carteolol, dipivefrin, dorzolamide, brinzolamide, latanoprost, unoprostone, and pilocarpine on intracellular free Ca2+ ([Ca2+]i) mobility in cultured bovine corneal endothelial cells. Various antiglaucoma drugs were diluted from original concentrations to 1/100, 1/1,000, and 1/10,000. The [Ca2+]i mobility was studied by spectrofluorophotometry after loading with the ester of fura-2 (fura-2/AM). It was found that timolol (58 μM and 5.8 μM), levobunolol (171 μM, 17.1 μM, and 1.71 μM), betaxolol (162 μM, 16.2 μM, and 1.62 μM), carteolol (680 μM and 68 μM), dipivefrin (28 μM and 2.8 μM), dorzolamide (616 μM and 61.6 μM), brinzolamide (260 μM), latanoprost (1.1 μM), unoprostone (28.2 μM, 2.82 μM, and 0.282 μM), and pilocarpine (408 μM and 40.8 μM) induced a significant increase in [Ca2+]i. Nevertheless, only brimonidine (68 μM and 6.8 μM) decreased [Ca2+]i concentration significantly. Benzalkonium chloride preservative did not affect [Ca2+]i after addition of 0.001, 0.0001 and 0.00001 mg/mL to cells. These results indicate that all antiglaucoma drugs may affect the physiologic function of corneal endothelial cells through change of [Ca2+]i.mobility

Effects of Antiglaucoma Drugs on Cellular Proliferation in Cultured Human Corneal Keratocytes

The purpose of this study was to investigate the effects of various antiglaucoma drugs, including timolol, betaxolol, carteolol, levobunolol, dipivefrin, and pilocarpine, on cellular proliferation in cultured human corneal keratocytes. Human corneal keratocytes were cultured with RPMI-1640 medium containing 10% fetal bovine serum. Antiglaucoma drugs were prepared from original concentrations to dilutions of 1/10, 1/100, and 1/1,000. After exposure to drugs for 100 minutes, cellular proliferation was estimated by [3H] thymidine uptake methodology. It was found that cellular proliferation in corneal keratocytes was inhibited by only a 1/10 dilution of various drugs including timolol, betaxolol, carteolol, levobunolol, dipivefrin, and pilocarpine. The [3H]thymidine uptake values were significantly inhibited to 63%, 18%, 87%, 68%, 55%, and 67% by a 1/10 dilution of the above drugs. However, the cellular proliferation was also significantly suppressed by 0.01 mg/mL of benzalkonium chloride preservative. It is shown that the inhibition of cellular proliferation by high concentrations of antiglaucoma drugs may result from the benzalkonium chloride preservative contained in these drugs

Effects of Antibiotics and Corticosteroid Eyedrops on Cellular Proliferation in Cultured Human Corneal Keratocytes

The purpose of this study is to investigate the effects of antibiotics and glucocorticoid eyedrops, including gentamicin, sulfisomezole, fluorometholone, dexamethasone, and betamethasone, on cellular proliferation in cultured human corneal keratocytes. Human corneal keratocytes were cultured in RPMI-1640 containing 10% fetal bovine serum. Drugs were prepared from original concentrations to 1/10, 1/100, and 1/1,000 dilutions. After exposure to drugs for 100 minutes, cellular proliferation was estimated by [3H]-thymidine uptake. It was found that cellular proliferation in corneal keratocytes was not affected by any of the three dilutions of gentamicin but was inhibited by 1/10 and 1/100 dilutions of sulfisomezole to 82% and 90% of control. [3H]thymidine uptake values were inhibited to 75% by 1/10 dilution of fluorometholone and by 1/10 and 1/100 dilutions of betamethasone to 84% and 86% of control. Meanwhile, cellular proliferation was significantly inhibited by 1/10, 1/100, and 1/1,000 dilutions of dexamethasone to 82%, 86%, and 90%, respectively, in comparison with control values. It was demonstrated that commercial eyedrops of glucocorticoids inhibit cellular proliferation in corneal keratocytes, which may modulate the wound healing of corneal stroma

Mechanism of 5-Fluorouracil-Induced Apoptosis on Cultured Corneal Endothelial

Repeated subconjunctival injections with 5-fluorouacil (5-FU) after trabeculectomy are widely used in glaucoma patients for the inhibition of excess scar formation in wound site. The aim of this study was to evaluate the toxic effects of 5-FU and mechanisms of drug-induced apoptosis in cultured porcine corneal endothelial cells. Cellular damage and the caspase pathway were estimated with a MTT assay. The apoptotic characteristics were detected with flow cytometry, a TUNEL test and Western blotting. The results indicated that 5-FU was toxic to corneal endothelial cells in a time- and dose-dependent manner. Pretreatment with a general caspase inhibitor, Z-VAD-FMK, a caspase-8 inhibitor, Z-IETD-FMK, and a caspase-9 inhibitor, Z-LEHD-FMK, reversed 5-FU-induced cellular damage. Following exposure to 5-FU, a flow cytometric assay with MitoLight dye demonstrated the loss of mitochondrial membrane potential. A positive TUNEL test revealed that cellular DNA apoptosis occurred following exposure to 0.05, 0.1, and 0.5 mg/ml 5-FU for 15 h. Annexin V-FITC and negative propidium iodide (PI) staining indicated that the cell membrane underwent apoptosis upon exposure to 0.1 and 0.5 mg/ml 5-FU for 15 h. The Western blot assay demonstrated up-regulation of the Bax, p53 and p21 proteins induced by 5-FU. Taken together, thes

Glaucomatocyclitic Crises May Occur in Patients with Narrow or Closed Angles

Purpose. To report cases of glaucomatocyclitic crises and discuss the possibility of occurrence in patients with narrow or closed angles. Background. The prevalence of angle closure is much higher among Asians than among the Western population. Currently, there is no evidence for a direct relationship between the etiology and angle structure. Design. A retrospective and observational case series. Methods. We retrospectively collected data from nine adult patients (three males and six females) who were diagnosed with a glaucomatocyclitic crisis and a shallow anterior chamber over a 21-year period, from 1995 to 2016, at the Kaohsiung Medical University Hospital. A narrow angle was defined as a grade less than the Shaffer system grade II. Ophthalmic examinations, including anterior segment biomicroscopy, direct ophthalmoscopy, intraocular pressure measurements, anterior chamber reaction, visual field tests, and the grade of the anterior chamber angle according to the Shaffer system, were reviewed. Results. These patients experienced at least one typical unilateral ocular hypertensive episode that fulfilled the criteria of a glaucomatocyclitic crisis without the angle feature. All patients had gonioscopically narrow or closed angles with or without peripheral anterior synechiae. Conclusions. The coexistence of narrow or closed angles and a glaucomatocyclitic crisis is possible, especially in patients of Asian descent. In patients with shallow anterior chambers, a glaucomatocyclitic crisis may be a cause of acute glaucoma episodes

Dexamethasone Intravitreal Implant (Ozurdex) for Long-Term Macular Edema after Epiretinal Membrane Peeling Surgery

Purpose. To investigate the functional and anatomical outcome of the 0.7 mg dexamethasone (Ozurdex) intravitreal implant (IVD) in eyes with long-term macular edema after macular epiretinal membrane removal. Methods. We enrolled 40 eyes with persistent macular edema at least 12 months after epiretinal membrane removal. Twenty eyes in the IVD group received IVD and the other 20 eyes were in the control group. The main outcome measures were change in best-corrected visual acuity (BCVA) and central foveal thickness (CFT). Results. For eyes in the IVD group, the mean BCVA improved by 3.45 lines to 0.47 logMAR one month after IVD. However, the mean BCVA improved by only 0.14 lines to 0.74 logMAR at the same time in eyes in the control group. Six months later, the mean BCVA improved to 0.31 and 0.74 logMAR in the IVD and control groups, respectively. In the IVD group, the mean CFT decreased rapidly by 116.8 μm to 333.9 μm one month after IVD. Thereafter the CFT decreased at a slower pace. In the control group, the CFT remained static during the follow-up period. However, in the IVD group, 6 months after IVD, the CFT seemed to have a tendency to increase. Conclusions. Single IVD could significantly decrease macular edema and improve visual outcome for eyes with persistent long-term macular edema after macular ERM removal and the effect can be sustained as long as 6 months after the initial injection. However, in order to maintain the visual and anatomical outcome, repeat IVD might be considered if macular edema recurs

Comparison of Visual Outcome and Morphologic Change between Different Surgical Techniques in Idiopathic Epiretinal Membrane Surgery

Purpose. To investigate the morphological and functional outcomes of idiopathic epiretinal membrane (ERM) surgery between three different surgical techniques: ERM peeling only, whole-piece ILM peeling, and maculorrhexis ILM peeling. Patients and Methods. This is a retrospective, consecutive, and comparative study enrolling 60 patients from Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. Surgery performed between July 2011 and June 2012 was done with ERM peeling only (group I). ERM peeling and ILM peeling as a whole piece (group II) were performed between July 2012 and July 2013. Surgery performed between August 2013 and December 2014 was done with maculorrhexis ILM peeling (group III). Main outcome measures include visual acuity change (BCVA) and central foveal thickness (CFT). Results. At 12 months postoperation, the mean BCVA in group III was significantly better than in group I and group II. Comparison of CFT reduction between the three groups revealed significantly more reduction in group III than in group II at all postoperative follow-up periods. Eyes with restoration of foveal depression were observed in 52.6% in group I, 52.4% in group III, but only 20% of eyes in group II. None of the eyes in both ILM peeling groups encountered recurrence of macular pucker formation. Conclusion. All three techniques can achieve visual acuity improvement and macular thickness reduction. Maculorrhexis ILM peeling achieves more rapid improvement of visual function, better final visual outcome, and a higher rate of normal foveal contour than whole-piece ILM peeling