Comparison of cytokine expression and ultrastructural alterations in fresh-frozen and dried electron beam-irradiated human amniotic membrane and chorion.

The purpose of the current study was to compare the effects of drying and fresh-freezing on human amniotic membrane (HAM) and amnion/chorion membrane (HACM) in terms of histological and structural characteristics and cytokine levels. HAM and HACM samples, obtained from six placentae, were investigated. HAM and HACM were dried, electron beam-irradiated (dehydration group; d-HAM/d-HACM), or fresh-frozen (freezing group; f-HAM/f-HACM). Luminex assay was used to assay the levels of 15 cytokines. The ultrastructural characteristics of HAM and HACM were evaluated using light and transmission electron microscopies. Total cytokine contents did not show the statistical difference between dehydration and fresh-freezing process. Significantly higher levels of total cytokines were observed in HACM than in HAM. Epidermal growth factor (EGF) level was significantly higher in d-HAM than in the other samples. The levels of most of the other growth factors were higher in HACM than in HAM, but there was no statistical difference between the dehydration process and the fresh-freezing process. The levels of the cytokines, other than the growth factors, were higher in HACM than in HAM, and higher concentrations of cytokines were observed in the freezing group than in the dehydration group. Histological examination revealed that the dehydration group had thinner tissues than the freezing group, but the structural stability, including the basement membrane, did not differ between the two groups. Microscopic structures such as microvilli and nuclei were well-preserved in the freezing group, based on the results of the transmission electron microscopy. Our dehydration process maintained the histological structure of HAM/HACM and a variety of growth factors and cytokines were identified. Especially, the HAM, processed with the dehydration method, had a higher EGF level than that processed with the fresh-freezing method. Therefore, dehydration method can be used to effectively promote wound repair.

Crossover clinical trial of pain relief in cataract surgery.

PURPOSE: To determine the effects of intra-operative Korean traditional music on pain experienced by Korean patients undergoing sequential bilateral cataract surgery. METHODS: This was a two-sequence, two-period, and two-treatment crossover study. Fifty-two patients with cataracts were divided into two groups by block randomization, and bilateral cataract surgery was performed. In group 1, patients listened to Korean traditional music (KTM) during their first but not second cataract surgery. This sequence was reversed for patients in group 2. After each surgery, patients scored their pain intensity (PI) using a visual analog scale (VAS) ranging from 0 to 10, where 0 was 'no pain' and 10 was 'unbearable pain.' RESULT: There was a statistically significant reduction in the mean VAS score with KTM (3.1 ± 2.0) compared to that without KTM (4.1 ± 2.2; p = 0.013). However, there were no statistically significant differences in blood pressure or pulse rates. CONCLUSION: KTM had a significant effect on reducing pain experienced by patients during cataract surgery. This may be useful in the context of other surgical procedures to reduce pain in Korean patients.

Direct visualization of aqueous tear secretion from lacrimal gland.

PURPOSE: To introduce a new method for direct observation of tear secretion from the palpebral lobes of the main lacrimal glands and Wolfring glands using fluorescein dye. METHODS: Twenty-six patients with dry eye and 26 healthy subjects were included in the study. Fluorescein was applied to the conjunctiva overlaying the palpebral lobe of the main lacrimal glands. Under cobalt blue light, we captured tear secretion from the glands in movies. We compared the number of excretory openings, the location and tear flow rate between the dry eye and healthy-subject groups. To observe tear secretion from the Wolfring glands, we everted the upper lids and applied fluorescein dye to the upper margin of the tarsus. RESULTS: Mean number of excretory openings on the palpebral lobe was 2.5 in the dry eye group and 3.4 in the healthy-subject group (p = 0.031). Most of the openings were observed at the ridge, the most protruding portion of the palpebral lobe. Mean flow rate from the palpebral lobe was 0.45 µl/min in the dry eye group and 0.91 µl/min in the healthy-subject group (p = 0.046). Mean flow rate from the Wolfring glands was 0.007 µl/min in the dry eye group and 0.009 µl/min in the healthy-subject group (p = 0.615). CONCLUSION: This visualization method allowed us to directly observe tear secretion from the palpebral lobe of the main lacrimal glands and Wolfring glands, and to quantitatively measure the tear flow rate from the lacrimal glands.

Effects of Combined Photodynamic Therapy and Topical Bevacizumab Treatment on Corneal Neovascularization in Rabbits.

PURPOSE: To evaluate the effects of combined photodynamic therapy (PDT) and topical bevacizumab treatment on corneal neovascularization in rabbits. METHODS: Corneal neovascularization was induced by placing a suture for 7 days in both eyes of 15 rabbits. After suture removal, the right eyes of the rabbits were divided into 3 treatment groups (n = 5 each): group 1, 5 mg/mL bevacizumab eye drops; group 2, PDT at 150 J/cm; and group 3, combined PDT and topical bevacizumab treatment. All 15 left eyes formed a control group. After 2 weeks of treatment, digital photographs were obtained and analyzed to determine the area of corneal neovascularization. The rabbits were then killed to measure vascular endothelial growth factor (VEGF) levels in corneal tissues. RESULTS: The corneal neovascularization area after treatment was significantly smaller in the 3 treatment groups compared with that in the control group. A significant difference was observed between groups 1 and 2 and group 3, but not between groups 1 and 2. VEGF levels were significantly lower in groups 1 and 3 than in the control group. Although group 2 exhibited relatively higher VEGF levels compared with the control group, the difference was not significant. CONCLUSIONS: Our results suggest that combined PDT and topical bevacizumab treatment is significantly more effective than monotherapy for the treatment of corneal neovascularization in rabbits. Such quantitative comparison in the laboratory is believed to provide the theoretical basis for clinical application of combined PDT and topical bevacizumab treatment.

Calpain and Caspase-12 Expression in Lens Epithelial Cells of Diabetic Cataracts.

PURPOSE: To compare the expression of calpain and caspase-12 in human lens epithelial cells (LECs) of cataract patients with or without diabetic retinopathy (DR). DESIGN: Prospective, comparative case series. METHODS: Patients were divided into 4 groups: patients without diabetes mellitus (DM) (Group 1), patients with DM and without diabetic retinopathy (DR) (Group 2), diabetic patients with nonprolifeative DR (NPDR) (Group 3), and diabetic patients with proliferative DR (PDR) (Group 4). Data on DM duration and glycated hemoglobin (HbA1c) level were collected. The LECs were obtained during cataract surgery and immunohistochemical staining was performed for calpain and caspase-12. The reactivity score of each specimen was determined using confocal microscopy. RESULTS: A total of 40 patients (40 eyes) were evaluated, and each group was composed of 10 patients. The expression of calpain and caspase-12 was highest in Group 4, followed by Group 3, Group 2, and Group 1 (P < .001). The expressions were significantly higher with a longer duration of DM, increased HbA1c level, or advanced DR. CONCLUSIONS: Endoplasmic reticulum stress may be associated with apoptosis of LECs, resulting in cataract formation in diabetic patients.

The Effect of Topical Diquafosol Tetrasodium 3% on Dry Eye After Cataract Surgery.

PURPOSE: To evaluate the effectiveness of 3% diquafosol tetrasodium for treating dry eye after cataract surgery. METHODS: Among patients who underwent bilateral cataract surgery, 34, who met the diagnostic criteria for dry eye syndrome 1 week postoperatively, were enrolled. Patients were randomly assigned to receive 3.0% diquafosol tetrasodium ophthalmic solution in one eye and 0.9% saline in the other eye four times daily for 8 weeks. Dry eye severity was measured at 1, 5, and 9 postoperative weeks using the Schirmer 1 test (SIT), tear film breakup time (TBUT), and fluorescein corneal staining. tear meniscus height (TMH), tear meniscus depth (TMD), and tear meniscus area (TMA) measured using Fourier-domain optical coherence tomography and symptom questionnaire scores. RESULTS: TBUT and corneal staining significantly improved 8 weeks postoperatively in eyes treated with 3.0% diquafosol tetrasodium (p < 0.01, p < 0.01) and were better than normal saline-treated eyes (p < 0.01, p < 0.01). SIT did not improve (p = 0.26). TMH, TMD, and TMA did not improve at 4 and 8 weeks. All symptom questionnaire scores improved in eyes treated with 3.0% diquafosol tetrasodium (all p < 0.01). CONCLUSION: The 3.0% diquafosol tetrasodium treatment improved tear film stability and subjective symptoms of dry eye after cataract surgery. Increased mucin production as a result of diquafosol treatment may have caused these results.

A case of corneal cystinosis in a patient with rickets and chronic renal failure.

A 22-year-old man diagnosed with nephropathic cystinosis at the age of 4 years was found to have progressive bilateral corneal crystal deposition. He presented with severe photophobia and decreased visual acuity. Ocular cystinosis was diagnosed on observing the typical crystals. Optical coherence tomography showed multiple areas of stromal hyperreflectivity due to crystal deposits within the corneal stroma. Ex vivo transmission electron microscopy of the cornea showed pathognomonic crystal deposits in corneal stromal keratocytes. Using polymerase chain reaction sequencing of the entire coding region, we identified five gene mutations, including two unreported mutations.

Assessment of the Tear Meniscus Using Optical Coherence Tomography in Patients With Type 2 Diabetes Mellitus.

PURPOSE: To analyze tear meniscus dimensions with Fourier domain-optical coherence tomography (FD-OCT) in patients with type 2 diabetes mellitus. METHODS: Patients with type 2 diabetes without diabetic retinopathy (DR) (n = 54), with nonproliferative DR (n = 45), with proliferative DR (PDR) (n = 25), and controls (n = 45) were enrolled. Hemoglobin A1c and Early Treatment Diabetic Retinopathy Study (ETDRS) stages of DR were evaluated. Schirmer test (ST), tear breakup time test, and fluorescein corneal staining were carried out. Tear meniscus height (TMH), tear meniscus depth (TMD), and tear meniscus area (TMA) were obtained by FD-OCT. RESULTS: TMH, TMD, and TMA in the diabetes and control groups were 233 ± 79 µm, 148 ± 53 µm, and 0.026 ± 0.015 mm, and 261 ± 67 µm, 167 ± 49 µm, and 0.032 ± 0.017 mm, respectively. All parameters were lower in diabetes (all P < 0.01). And the values in PDR were 185 ± 90 µm, 117 ± 50 µm, and 0.017 ± 0.010 mm which were lower than those in control and nonproliferative DR (227 ± 71 µm, 144 ± 50 µm, and 0.023 ± 0.010 mm) (all P < 0.01). TMH, TMD, and TMA correlated with ST (P < 0.01, P = 0.02, and P < 0.01, respectively) and the corneal staining score (all P < 0.01). Patients with longer diabetes duration and a history of panretinal photocoagulation showed significantly decreased TMH, TMD, and TMA (P < 0.01). CONCLUSIONS: OCT, along with tear breakup time and ST, is an effective way to assess tear function in type 2 diabetes. Patients with PDR and a history of panretinal photocoagulation showed lower tear meniscus parameters. The decrease in tear meniscus parameters was correlated with diabetes duration.

Effect of diabetic retinopathy and diabetes on the intraocular straylight in pseudophakic eyes.

BACKGROUND: The aim of this study was to prove the relationship between the intraocular straylight level and diabetic retinopathy (DR) according to disease severity. Also, we aimed to evaluate whether diabetes mellitus (DM) per se could be a risk factor of increased intraocular straylight although we did not rely on a definite sign of DR in this study. METHODS: In this prospective comparative study, ninety three eyes were enrolled and divided into four groups as follows: Group 1 (26 eyes), without DR or DM; Group 2 (25 eyes), with DM but without DR; Group 3 (21 eyes), mild to moderate non-proliferative DR; and Group 4 (21 eyes), severe non-proliferative DR. To measure the intraocular straylight in an objective manner, the C-quant straylight meter was used to preoperatively and 2 months postoperatively in all patients who underwent phacoemulsification surgery. All the patients also underwent a macular optical coherence tomography and hemoglobin A1c (HbA1c) analysis. A comparison of straylight levels adjusted by age among four groups was performed postoperatively. RESULTS: The postoperative level of intraocular straylight was statistically significantly different among four groups (P <0.05). When adjusted for ages, Group 4 showed the highest straylight level when compared with Group 3 and the other two groups (P <0.05). Group 1 showed the lowest straylight level in comparison with Group 2 and the other two groups (P <0.05). There was no significant correlation between HbA1c level, duration of diabetes and postoperative straylight level. CONCLUSIONS: The level of intraocular straylight at 2 months postoperatively had a tendency to increase as the severity of DR increased. Additionally, the straylight level was higher in DM patients without DR than in patients without DM. Therefore, the severity of DR seemed to influence the intraocular straylight level. Although there is no definite sign of DR, DM per se can be a risk factor for increasing intraocular straylight. In conclusion, the level of intraocular straylight seems to be a sensitive test for detecting early retinal damage secondary to DM.

Comparison of Tear Meniscus Height Measurements Obtained With the Keratograph and Fourier Domain Optical Coherence Tomography in Dry Eye.

PURPOSE: To study the repeatability and reproducibility of tear meniscus height (TMH) measurements obtained with a keratograph and Fourier domain optical coherence tomography (FD-OCT) and to assess their agreement in patients with dry eye. METHODS: Sixty-four eyes with dry eye were analyzed by the Schirmer test, tear breakup time test, and fluorescein corneal staining. The TMH was measured 3 times using both devices by 2 different examiners. The repeatability and reproducibility of measurements were assessed by within-subject standard deviation (Sw), repeatability (2.77 Sw), coefficient of variation (CoV), and intraclass correlation coefficient. RESULTS: The TMH measured with the keratograph and FD-OCT was 0.232 ± 0.074 mm and 0.308 ± 0.129 mm, respectively (P < 0.01). A close correlation was found between the TMH measured with the keratograph and FD-OCT (r = 0.343). There was a negative correlation between the mean TMH and differences in the TMH measured with the keratograph and FD-OCT (r = 0.359). Both measurements correlated with the Schirmer test score, tear breakup time, and corneal staining score with P < 0.01. Intraexaminer CoV, 2.77 Sw, and intraclass correlation coefficient of the TMH were <6.5%, <0.059 mm, and >0.986, respectively, and interexaminer CoV and 2.77 Sw were 5.58% and 0.039 mm, respectively. CONCLUSIONS: Although the TMH measured with the keratograph tended to be lower than that measured with FD-OCT, the TMH measured with the keratograph closely correlated with the TMH measured with FD-OCT and conventional tests and had good repeatability and reliability.

Inhibitory Effect of Topical Aflibercept on Corneal Neovascularization in Rabbits.

PURPOSE: To examine the inhibitory effect of topical aflibercept [vascular endothelial growth factor (VEGF) trapR1R2] on corneal neovascularization (NV) in rabbits. METHODS: Corneal NV was induced in 24 eyes of 12 rabbits. Seven days after a silk suture in the corneal stroma, the rabbits were divided into 4 groups of 6 eyes each. Two groups were treated with topical aflibercept at 2 different concentrations: 2 mg/0.5 mL (0.1%, group 1) and 2 mg/5 mL (0.01%, group 2). The other 2 groups were treated with topical bevacizumab 2.5 mg/1 mL (0.1%, group 3) and topical balanced salt solution (group 4, control). The concentration of VEGF and placental growth factor (PIGF) messenger RNA (mRNA) was measured by reverse transcriptase-polymerase chain reaction. RESULTS: The surface area of NV was significantly smaller in the treatment groups compared with that of the control group. The expression of VEGF mRNA was 0.227 in 0.1% aflibercept (group 1), 0.811 in 0.01% aflibercept (group 2), and 0.495 in 0.1% bevacizumab (group 3). There was a significant decrease in the VEGF concentration in all 3 treatment groups compared with the control group, 1.491 (P = 0.031, <0.05). In the 0.01% aflibercept group, the difference was less than that of the 0.1% aflibercept and 0.1% bevacizumab groups. There was no significant difference in the 0.1% aflibercept and 0.1% bevacizumab groups. The expression of PIGF mRNA was 0.791 in 0.1% aflibercept (group 1), 0.743 in 0.01% aflibercept (group 2), 1.194 in 0.1% bevacizumab (group 3), and 1.458 in the control group. The expression of PIGF mRNA was significantly decreased in the 0.1% aflibercept and 0.01% aflibercept groups. CONCLUSIONS: Topical aflibercept may have an inhibitory effect on corneal NV in rabbits.

Comparative study of tacrolimus and bevacizumab on corneal neovascularization in rabbits.

PURPOSE: To compare the antiangiogenic effects of tacrolimus and bevacizumab on corneal neovascularization (CNV) in rabbits. METHODS: Neovascularization was induced in 32 eyes of 16 rabbits by placing a suture in the corneal stroma. Seven days after suture placement, all rabbits were divided into 4 groups and were treated subconjunctivally with bevacizumab (AVA_sub) 0.05 mL (5 mg/0.05 mL), tacrolimus (TAC_sub) 0.05 mL (0.25 mg/0.05 mL), balanced salt solution (0.05 mL was subconjunctivally injected in 1 eye of each rabbit and applied by eye drops in the other eyes, control group), and tacrolimus eye drops (TAC_drop) (5 mg/5 mL applied 4 times daily). Digital photographs were obtained and surface area of CNV was measured 7 days after subconjunctival injections. Corneal specimens were analyzed histopathologically and were used to measure the concentration of vascular endothelial growth factor (VEGF), tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin-1 beta (IL-1ß), and monocyte chemoattractant protein 1 mRNA by reverse transcription polymerase chain reaction. RESULTS: In digital photographs, the neovascularized area was decreased in all treatment groups (AVA_sub, 0.58; TAC_sub, 0.60; TAC_drop, 0.68) compared with the control group (balanced salt solution, 0.81). Histological examination showed markedly regressed new vessels in treatment groups, and immunohistochemical staining revealed weakly stained anti-VEGF and anti-F4/80 antibodies in treatment groups. In semiquantitative reverse transcription polymerase chain reaction, concentration of VEGF (AVA_sub, 0.24; TAC_drop, 0.18), TNF-α (AVA_sub, 0.19; TAC_sub, 0.24; TAC_drop 0.15), and IL-1ß (AVA_sub, 0.19; TAC_sub, 0.33; TAC_drop, 0.18) mRNA were significantly lower in treatment groups than in the control group (VEGF, 0.47; TNF-α, 0.44; IL-1ß, 0.87) (P < 0.05). CONCLUSIONS: Topical and subconjunctival tacrolimus application may be useful in reducing CNV and have comparable effects to subconjunctival bevacizumab injection.

Topographical Analysis of Non-Glaucomatous Myopic Optic Discs Using a Confocal Scanning Laser Ophthalmoscope (TopSS).

OBJECTIVES: To show normative data of optic discs and the mechanism of glaucoma in people with myopia. DESIGN: Cross-sectional study. PARTICIPANTS: This study investigated 89 Korean adults with myopia but without glaucoma. METHODS: Patients were divided into three groups according to the refractive error: low, moderate, and high; and axial length: normal or below normal length, moderately long, and extremely long. Optic disc variables were obtained by confocal scanning laser ophthalmoscope and compared among groups. RESULTS: The optic disc parameters have a correlation between the refractive error and the optic disc parameters such as average depth, volume below, and half-depth volume. Those parameters also decreased as the axial length increased. The thickness of the volume above decreased significantly as the axial length increased, but a similar relationship was not evident with the refractive error change. In addition, the optic disc parameters were analyzed with respect to the 12 clockwise directions. CONCLUSIONS: Analyses of optic disc parameters provided by TopSS™ revealed the height of the disc decreased as the myopic refractive error and/or axial length increased. The RNFL bundle became compacted in the thinner disc of the myopic population. This could be an explanation for the fragility of the RNFL in the myopic population. The 12 radial section analyses revealed the shallow cupping at the temporal side in the high-myopic, very-long-axis group. The neuroretinal rim (NRR) height significantly decreased at the superior and inferior sides. These findings suggest that the RNFL bundle should be under high mechanical strain in these sectors.

Optical quality in central serous chorioretinopathy.

PURPOSE: To assess optical quality and intraocular scattering using the Optical Quality Analysis System (OQAS) in central serous chorioretinopathy (CSC) and to determine the effects of retinal changes on optical quality. METHODS: This was a prospective, case-control study. Participants were 29 patients with diagnosis of CSC. The control group consisted of the patients' unaffected eyes. Initial logMAR visual acuity, central macular thickness (by spectral domain optical coherence tomography), and optical quality parameters including modulation transfer function (MTF) cutoff frequency, Strehl (2-dimensional) ratio, and OQAS values at 100%, 20%, and 9% contrast levels were investigated. Objective scattering index (OSI) at 4.0-mm pupil size was assessed in both eyes by using the OQAS. After 3 months of treatment, which included observation and focal laser or injections of antivascular endothelial growth factor, every CSC-affected eye was followed. Main outcome measures were differences between clinical parameters of the CSC-affected eye and those of the control eye and changes in those parameters according to the clinical course of CSC over 3 months. RESULTS: In CSC-affected eyes, the MTF cutoff was significantly reduced (P = 0.01), and OSI was significantly increased (P = 0.03). As macular thickness decreased, OSI decreased but did not become normalized compared to the control eye, nor was it statistically significantly correlated with central macular thickness change. CONCLUSIONS: Retinal change affected optical quality and intraocular scatter. Therefore, when the severity of a cataract is assessed using the OQAS, retinal status should be considered when interpreting OQAS values.

Effects of subconjunctival tocilizumab versus bevacizumab in treatment of corneal neovascularization in rabbits.

PURPOSE: The aim of this study was to compare the antiangiogenic effects of subconjunctival application of bevacizumab and tocilizumab on the regression of corneal neovascularization (NV) in rabbits. METHODS: Corneal neovascularization was induced in 48 eyes of 24 rabbits. Seven days after suture placement, the rabbits were divided into 4 groups of 6 rabbits each and treated subconjunctivally with 0.1 mL balanced salt solution (group 1), 0.1 mL tocilizumab (0.25 mg per 0.1 mL and 2.5 mg per 0.1 mL, groups 2 and 3), or 0.1 mL bevacizumab (2.5 mg per 0.1 mL) (group 4). Digital photographs of the eyes were obtained and the surface areas of corneal neovascularization were measured on days 7 and 14 after subconjunctival injections. On days 7 and 14, 3 rabbits were randomly chosen and the eyes were extracted. Half of the corneal specimens were analyzed histopathologically, and the other half were used to measure the concentrations of vascular endothelial growth factor (VEGF) and IL-6 using a multiplex bead assay, and the levels were compared with those of the controls. RESULTS: The surface areas of induced corneal neovascularization were significantly smaller in groups 3 and 4 (2.5 mg of tocilizumab and 2.5 mg of bevacizumab) compared with the control group on days 7 and 14 (P < 0.05). Group 2 did not show significant difference from the control group on days 7 and 14. There were no differences observed in the reduced neovascularization areas in groups 3 and 4 on days 7 and 14. The concentrations of VEGF in groups 3 and 4 were significantly lower than in the control group, and IL-6 mRNA levels were significantly lower in group 3 than in the other groups (P < 0.001) on days 7 and 14. Immunohistochemical analysis confirmed the reduced expression of VEGF in all 3 experimental groups compared with the control group. CONCLUSIONS: An antiangiogenic effect was observed after subconjunctival injection of 2.5 mg tocilizumab to an extent similar to that seen with 2.5 mg bevacizumab, which indicates that subconjunctival application of tocilizumab is effective for the inhibition of corneal neovascularization.

Toxicity of intrastromal voriconazole injection on corneal endothelium in rabbits.

PURPOSE: The aim of this study was to evaluate the toxicity of intrastromally injected voriconazole on corneal endothelial cells in rabbits. METHODS: In total, 32 eyes of 16 rabbits (8 eyes for each group) were divided into 4 groups according to the concentration of voriconazole [group A: 50 µg/0.1 mL (0.05%), group B: 100 µg/0.1 mL (0.1%), group C: 250 µg/0.1 mL (0.25%), or group D: 500 µg/0.1 mL (0.5%)]. Right eyes were injected intrastromally with voriconazole at concentrations of 50 µg/0.1 mL, 100 µg/0.1 mL, 250 µg/0.1 mL, or 500 µg/0.1 mL. Left eyes were injected intrastromally with isotonic saline as controls. Central corneal thickness and endothelial cell counts were measured before and at 6 hours, 1 day, and 1 week after the injection was given. Corneas were then harvested for transmission electron microscopy. RESULTS: Only the 0.5% group did not significantly recover from corneal edema 1 week after the injection (P = 0.167, P = 0.051, P = 0.086, P = 0.001 in groups A-D, respectively). There were significant differences in endothelial cell counts for the 0.1% and 0.25% groups (P = 0.077, P = 0.019, P = 0.008 in groups A-C, respectively). Transmission electron microscopy evaluation revealed definite necrotic changes in endothelial cells at concentrations of 0.1%, 0.25%, and 0.5%, but only microstructural changes at a concentration of 0.05%. CONCLUSIONS: Our results suggest that voriconazole could be injected safely into the intrastromal layer at a concentration of 0.05% with low endothelial cell toxicity. However, injections should be administered with caution because of the risk of microstructural damage.

Expression of p53 and caspase-8 in lens epithelial cells of diabetic cataract.

PURPOSE: To determine expression of apoptotic factors p53 and caspase-8 in human lens epithelial cells (LECs) of cataract patients with or without diabetic retinopathy (DR), the duration of diabetes mellitus (DM), and the level of glycated hemoglobin (hemoglobin A1c [HbA1c]). SETTING: St. Mary's Hospital, Catholic University of Korea, Seoul, South Korea. DESIGN: Randomized prospective study. METHODS: The LECs were isolated during cataract surgery. The isolated samples were classified into 4 groups as follows: patients without DM (Group 1), patients with DM but not DR (Group 2), diabetic patients with nonproliferative DR (Group 3), and diabetic patients with proliferative DR (Group 4). To explore the mechanism of apoptosis, the expressions of p53 and caspase-8 were measured by immunohistochemical staining and compared with the data according to the duration of DM, HbA1c levels, and severity of DR. RESULTS: All groups comprised 15 eyes. The expressions of P53 and caspase-8 were higher in Groups 2, 3, and 4 than in Group 1 (P<.001). The expressions were statistically significantly increased with a longer duration of DM, higher HbA1c levels, and advanced DR. CONCLUSIONS: The expressions of P53 and caspase-8 were strong in patients with DM and advanced DR. Knowledge of these relationships may lead to a better understanding of the development of diabetic cataract. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Ranibizumab injection for corneal neovascularization refractory to bevacizumab treatment.

Vascular endothelial growth factor inhibitor is an emerging therapeutic modality for various ocular diseases with neovascularization (NV). However, for corneal NV, controversy remains regarding whether bevacizumab or ranibizumab is superior. A 32-year-old female diagnosed with herpetic keratoconjunctivitis with refractory corneal NV despite two previous subconjunctival and intrastromal bevacizumab injections, received two subconjunctival and intrastromal ranibizumab injections. Six months postoperatively, there was significant regression of the neovascular area and vessel caliber. Here, the authors report a case of improvement in corneal NV with subconjunctival and intrastromal ranibizumab injections, which was previously refractory to bevacizumab injection. The findings may suggest a new prospect in treating corneal NV.

Effect of topical epigallocatechin gallate on corneal neovascularization in rabbits.

PURPOSE: The aim of this study was to evaluate the efficacy of topical application of epigallocatechin gallate (EGCG) for the treatment of corneal neovascularization in a rabbit model. METHODS: Corneal neovascularization was induced in 12 rabbits by placing a black silk suture in the corneal stroma (24 eyes) for a week. After suturing, 1 randomly chosen eye of the 12 rabbits was treated with topical EGCG at 2 different concentrations: 0.01% (group 1) and 0.1% (group 2), whereas the contralateral eyes were treated with sterilized balanced salt solution as the control. All eye drops were applied for 2 weeks after suturing. The suture materials were removed from all eyes on day 7. The surface area of corneal neovascularization was measured and analyzed in all eyes on days 7 and 14. On day 14, all eyes were extracted to measure the concentrations of vascular endothelial growth factor (VEGF) messenger RNA and cyclooxygenase-2 (COX-2) protein. RESULTS: The surface area of induced corneal neovascularization was significantly smaller only in group 2 compared with that of the control group on days 7 and 14 (P < 0.001). The change in surface area of corneal neovascularization after removal of the suture material was not significantly different between all 3 groups. VEGF messenger RNA levels were significantly lower in group 2 than in the control group (P < 0.001). The concentration of COX-2 was significantly lower in group 2 than in the control group (P = 0.043), but no significant difference was observed between group 1 and the control group. CONCLUSIONS: Topical administration of EGCG effectively inhibits corneal neovascularization in rabbits. This inhibitory effect is probably related to the suppression of VEGF and COX-2 meditated angiogenesis.