Macular holes following vitrectomy for rhegmatogenous retinal detachment: epiretinal proliferation and spontaneous closure of macular holes.

PURPOSE: To describe the characteristics and management of full-thickness macular holes (MHs) that develop after pars plana vitrectomy for rhegmatogenous retinal detachment (RD). METHODS: Retrospective, interventional, consecutive case series. Patients who developed secondary full-thickness MHs after prior pars plana vitrectomy for RD over a 6-year period were included. The main outcome measures included optical coherence tomography (OCT) findings and the clinical course of full-thickness MHs. RESULTS: A total of 11 eyes of 11 consecutive patients were included in the study. The mean age of the patients was 58.8 years (range, 47-70 years). The median time between RD repair and MH diagnosis was 36 months (range, 1 month-11 years). The fovea was attached to 10 eyes (91%) at the time of RD repair. OCT demonstrated epiretinal proliferation (EP) at the hole margin in 10 eyes (91%). MH spontaneously closed in 7 eyes (63%) but reopened in 5 eyes. A total of 7 eyes (63%) required a vitrectomy to repair the MHs. All MHs were closed at the last follow-up visit. CONCLUSION: Full-thickness MHs after pars plana vitrectomy for RD have features that are distinct from that of typical idiopathic MH. The presence of EPs is common, and MHs are prone to spontaneous closure and reopening. These findings suggest that EP may be associated with spontaneous hole closure and that long-term follow-up is necessary even if the MHs close spontaneously.

Structural Changes of Inner and Outer Choroid in Central Serous Chorioretinopathy Determined by Optical Coherence Tomography.

PURPOSE: To determine the structural changes of the choroid in eyes with central serous chorioretinopathy (CSC) by enhanced depth imaging optical coherence tomography (EDI-OCT). METHODS: A retrospective comparative study was performed at two academic institutions. Forty eyes with CSC, their fellow eyes, and 40 eyes of age-matched controls were studied. Subfoveal cross sectional EDI-OCT images were recorded, and the hypo reflective and hyperreflective areas of the inner and outer choroid in the EDI-OCT images were separately measured. The images were analyzed by a binarization method to determine the sizes of the hyporeflective and hyperreflective areas. RESULTS: In the inner choroid, the hyperreflective area was significantly larger in the CSC eyes (35,640±10,229 µm2) than the fellow eyes (22,908±8,522 µm2) and the control eyes (20,630±8,128 µm2; P<0.01 vs control for both, Wilcoxon signed-rank test). In the outer choroid, the hyporeflective area was significantly larger in the CSC eyes (446,549±121,214 µm2) than the control eyes (235,680±97,352 µm2, P<0.01). The average ratio of the hyporeflective area to the total choroidal area was smaller in the CSC eyes (67.0%) than the fellow eyes (76.5%) and the control eyes (76.7%) in the inner choroid (P<0.01, both). However, the ratio was larger in the CSC eyes (75.2%) and fellow eyes (71.7%) than in the control eyes (64.7%) in the outer choroid (P<0.01, both). CONCLUSIONS: The larger hyperreflective area in the inner choroid is related to the inflammation and edema of the stroma of the choroid in the acute stage of CSC. The larger hyporeflective areas in the outer choroid is due to a dilatation of the vascular lumens of the larger blood vessels. These are the essential characteristics of eyes with CSC regardless of the onset.

Responsiveness of eyes with polypoidal choroidal vasculopathy with choroidal hyperpermeability to intravitreal ranibizumab.

BACKGROUND: To determine the role played by vascular endothelial growth factor (VEGF) in polypoidal choroidal vasculopathy (PCV) based on an interventional immunology theory. METHODS: Eyes with PCV were divided in a masked fashion into those with choroidal hyperpermeability (HP group) and those with normal choroidal permeability (NP group) based on the indocyanine green angiograms. The inter-rater agreement rate was evaluated using Fleiss' kappa. Patients were treated by intravitreal ranibizumab (IVB). The central choroidal thickness and central foveal thickness (CFT) at the baseline and 7 days after the treatment were measured by optical coherence tomography. RESULTS: Among the 57 consecutive eyes diagnosed with PCV, 42 eyes of 42 patients met the inclusion criteria (21 eyes/HP group vs 21 eyes /NP group). Central choroidal thickness in HP group was significantly thicker than that in the NP group (P < .001, Mann-Whitney U test). The inter-rater agreement was high with a Fleiss' kappa = 0.95, P < .0001. The percentage reduction in the CFT in HP group (14.0%) was significantly less than that in NP group (20.4%; P = .013, Mann-Whitney U test). CONCLUSIONS: Eyes with PCV that are associated with choroidal hyper-permeability may not be strongly associated with VEGF-related pathology, and may not respond favorably to anti-VEGF monotherapy.

TNF-α disrupts morphologic and functional barrier properties of polarized retinal pigment epithelium.

Retinal pigment epithelial (RPE) cells form a blood-ocular barrier, and their polarized property is crucial for maintaining the barrier functions. Tumor necrosis factor alpha (TNF-α), a major pleotropic inflammatory cytokine that disrupts the barrier function and eventual angiogenesis, is expressed in the choroidal neovascularizations of age-related macular degeneration eyes. Thus, it most likely plays an important role in the progression of the disease. The purpose of this study was to compare the effects of TNF-α on the barrier function of polarized RPE cells. Non-polarized RPE cells were used as negative controls. Isolated porcine RPE cells were seeded on Transwell™ membranes. The polarization of the RPE cells was determined by their high transepithelial electrical resistance (TER >150 Ω cm(2)) and by their differential secretion of vascular endothelial growth factor (lower layer/upper layer >2.5X). Polarized RPE cells were incubated with 10 ng/ml of TNF-α and the TER was measured. TNF-α significantly decreased the TER of polarized RPE cells by 17.6 ± 2.7% (P < 0.001) of the control at 24 h and that of non-polarized RPE cells by 5.4 ± 6.5% (P = 0.401). The p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, blocked the effects of TNF-α of decreasing the TER. Cell junction-related molecules, e.g., ZO-1, located between cells in control RPE cells, were disassembled by TNF-α, and this breakdown was suppressed by SB203580 in polarized RPEs. These results indicate that the breakdown of the RPE barrier function was caused exclusively by TNF-α in polarized RPEs, and TNF-α was acting through the p38 MAPK pathways. Investigations of polarized RPE cells should be more suitable for in vitro studies of the pathophysiology of retinochoroidal diseases.

Repeatability and reproducibility of subfoveal choroidal thickness in normal eyes of Japanese using different SD-OCT devices.

PURPOSE: To compare subfoveal choroidal thickness (SCT) measurements of three different commercially available spectral-domain optical coherence tomography instruments with healthy eyes of Japanese. METHODS: A prospective, cross-sectional study was performed at a single institution. SCT of the right eye of 43 normal subjects was measured using three different SD-OCTs: Heidelberg Spectralis-OCT (Spectralis), Cirrus HD-OCT (Cirrus), and Topcon 3D OCT-1000 Mark II (Topcon). Two separate measurements were performed for the same eye with a maximum by a single examiner. SCT was defined as the distance from the posterior edge of the retinal pigment epithelium to the choroid/sclera junction. After manual segmentation, measurements were made using calipers equipped on each machine by masked raters. Intraclass, interrater, and intermachine agreements were assessed. RESULTS: Forty-three subjects (mean age, 30.5 years) were enrolled. Of 43 eyes, the SCT of 39 eyes (90.7%) could be measured using each machine. Intraclass correlation coefficients (95% confidence intervals) were 0.976 (0.954-9.987), 0.958 (0.919-0.978), and 0.939 (0.895-0.971) with Spectralis, Cirrus, and Topcon, respectively. Interrater correlation coefficients (95% confident interval) were 0.944 (0.893 to 0.971), 0.956 (0.831 to 0.983), and 0.924 (0.825 to 0.964) with Spectralis, Cirrus, and Topcon, respectively. The average SCT was 272.6, 272.8, and 269.2 µm with Spectralis, Cirrus, and Topcon, respectively. The intermachine correlation coefficient was significantly high among the machines (P<0.001, Spearman), 0.97 (Spectralis-Cirrus), 0.96 (Cirrus-Topcon), and 0.98 (Topcon-Cirrus). Bland-Altman plot analysis showed no typical trend among the machines. CONCLUSIONS: SCT measurements obtained with three different SD-OCTs were highly correlated and could be used interchangeably. (http://upload.umin.ac.jp number, UMIN000005287.).

Comparative study of vitrectomy versus intravitreous triamcinolone for diabetic macular edema on randomized paired-eyes.

BACKGROUND: The present study was performed to compare the effects of pars plana vitrectomy (PPV) and single intravitreaous triamcinolone acetonide (IVTA) on diabetic macular edema (DME) in paired eyes. METHODS: Prospective comparative study on randomized paired-eyes was carried out at two hospitals. Forty eyes of 20 patients with bilateral DME were included. One randomly-selected eye was treated with PPV (PPV group), and the other eye was treated with IVTA (4 mg, IVTA group). The central macular thickness (CMT) measured by optical coherence tomography (OCT) and best-corrected visual acuity (BCVA) were monitored for 12 months after treatment. Changes from baseline and differences between groups were analyzed using a mixed model. RESULTS: At 1 and 3 months, CMT decreased significantly in the IVTA group compared to baseline (p < 0.0001 both), but CMT then increased gradually and no significant difference was found at 12 months (p = 0.90). In the PPV group, CMT decreased continuously and reached a significant level at 12 months (p < 0.0001). CMT of the IVTA group was significantly less than that of the PPV group at 1 month (p = 0.009); however, there was no significant difference at 3 months. Conversely, CMT was significantly less in the PPV group than in the IVTA group at 12 months (p = 0.0003). The changes of BCVA paralleled those of CMT, but no significant difference was detected between baseline BCVA and any time point. CONCLUSIONS: Despite the short-term improvement, DME recurred 6 months after IVTA, while it remained resolved after PPV. Although this study did not reveal a significant change of BCVA with either treatment, PPV resolved DME more effectively than IVTA at 1 year.

Intravitreous VEGF-A in eyes with massive vitreous hemorrhage.

PURPOSE/BACKGROUND: Although vascular endothelial growth factor (VEGF) is abundant in serum, the intraocular concentration of VEGF in eyes with massive vitreous hemorrhage (VH) is not well-known. The present study was conducted to elucidate the effects of a massive VH on intravitreous VEGF concentration. METHODS: Vitreous samples were obtained during vitrectomy: 12 samples from eyes with epiretinal membrane without diabetic retinopathy (DR), and nine samples from massive VH with no DR, such as age-related macular degeneration, rhegmatogenous VH, Terson's syndrome and macro-aneurysm rupture. Twelve samples were obtained from proliferative DR. VEGF was measured with an enzyme-linked immunosorbent assay (ELISA). Samples incubated with or without heparin were also examined for the release of VEGF binding to the vitreous body. The localization of VEGF and type II collagen in the vitreous was evaluated from immunohistochemistry. RESULTS: The concentration of VEGF was significantly higher in eyes with proliferative DR (821 ± 949 pg/ml) than in non-DR with massive VH (2.75 ± 7.5 pg/ml, P < 0.01, chi-square test) or non-DR with no VH (less than detectable level, P < 0.01, chi-square test) There was no statistically significant difference between eyes with massive VH and non-diabetic eyes without VH. Treatment with heparin did not significantly affect the concentration of vitreous VEGF. VEGF was localized mainly in the clot from the results of an immunohistochemical analysis. CONCLUSIONS: Even with a massive VH, diffusible VEGF does not increase significantly in the liquid phase and is principally present in a clot. VH alone should not be an indication for vitrectomy from the point of view of VEGF-related pathology.

NSAIDs inhibit neovascularization of choroid through HO-1-dependent pathway.

Intraocular neovascularization is the leading cause of severe visual loss and anti-vascular endothelial growth factor (VEGF) therapy is currently performed for choroidal neovascularization (CNV). Despite its potent anti-angiogenic effect, there are concerns about its long-term safety. Non-steroidal anti-inflammatory drugs (NSAIDs) are common therapeutic agents used for treating inflammatory diseases, and their anti-stress effects are attracting attention now. We studied the effects of topical NSAIDs on CNV, focusing on anti-stress proteins. Cultured retinal pigment epithelium (RPE) cells were treated with NSAIDs: bromfenac, indomethacin, or vehicle control. Transcription factor NF-E2-related factor 2 (Nrf2) and its downstream anti-oxidant protein heme oxygenase (HO)-1 were assessed using western blot and immunohistochemistry. As a result, NSAIDs induced translocation of Nrf2 into the nucleus and the robust expression of HO-1 in a dose- and time-dependent manner. Flow cytometric analysis revealed that bromfenac inhibited H(2)O(2)-induced apoptosis in cultured RPE cells. Next, we studied the effects of topical bromfenac on laser-induced CNV model in rat. The expressions of Nrf2 and HO-1, infiltrations of ED-1-positive macrophages at CNV lesions and size were analyzed. VEGF in the ocular fluid of these rats was also measured using enzyme-linked immunosorbent assay. Rats administered an inhibitor of HO-1 stannic mesoporphyrin (SnMP) were also studied. The results showed that topical bromfenac led to translocation of Nrf2 and induction of HO-1 in CNV lesions and that the number of infiltrating macrophages at the CNV lesion decreased. The sizes of CNV lesions were significantly smaller in bromfenac-treated rats than control CNV, and the effects were diminished by SnMP. VEGF increased in the ocular fluid after laser treatment and was inhibited by bromfenac and SnMP canceling these effects. NSAIDs inhibit CNV through the novel anti-stress protein HO-1-dependent pathway, indicating its potential therapeutic value for various intraocular angiogenic diseases including CNV.

Early change of central macular thickness after intravitreous triamcinolone or bevacizumab in diabetic macular edema or retinal vein occlusion.

PURPOSE: To evaluate the immediate changes after intravitreous triamcinolone acetonide or intravitreous bevacizumab in diabetic macular edema (DME). METHODS: A nonrandomized interventional study. Type 2 diabetic patients were included. Intravitreous triamcinolone acetonide (4 mg) was injected for 22 eyes with DME and IVB (1.25 mg) for 18 eyes with DME. The early time-dependent changes of central macular thickness were evaluated by optical coherence tomography before and from 1 hour to 1 month after intervention. Intravitreous bevacizumab was also tested in patients with retinal vein occlusion as a control of non-DME. Visual acuity was also examined. RESULTS: Compared with the baseline, central macular thickness of eyes with DME decreased significantly 1 hour after intravitreous triamcinolone acetonide (P < 0.05, Wilcoxon signed rank test), while it did not significantly until 24 hours after IVB. The decrease in central macular thickness was observed significantly from 3 hours after IVB in retinal vein occlusion (P < 0.05, Wilcoxon signed rank test), and it was more evident in retinal vein occlusion than DME after IVB. Visual acuity improved significantly in DME with intravitreous triamcinolone acetonide or IVB at 1 month (P < 0.01 and P < 0.05, respectively, Wilcoxon signed rank test). CONCLUSION: Factors responsive to triamcinolone acetonide, other than vascular endothelial growth factor, might play an important role in pathogenesis of DME compared with retinal vein occlusion. Although no conclusion can be drawn, immediate decrease in central macular thickness after intravitreous triamcinolone acetonide might indicate the possible involvement of a nongenomic pathway of triamcinolone acetonide action.

Ocular surface molecule after transconjunctival vitrectomy.

AIM: To evaluate ocular surface molecules after transconjunctival pars plana vitrectomy (PPV). METHODS: A total of 28 eyes of 28 patients who received PPV were examined. Tears (10µl) were collected before and after 20-gauge PPV or 23-gauge PPV. The concentrations of interleukin (IL)-1ß, IL-6, IL-8, IL-10, IL-12p70 and tumour necrosis factor (TNF)-α were measured. The concentrations of each cytokine before and after surgery were compared. The change ratios of each cytokine (post-/presurgery) between 20-gauge and 23-gauge PPV were compared. RESULTS: IL-1ß, IL-6, IL-8 increased after PPV (Wilcoxon rank sum test; IL-1 ß, p=0.009; IL-6, p<0.001; IL-8, p<0.001), while the other cytokines did not. Among them, only IL-8 increased significantly in 20-gauge PPV than in 23-gauge PPV (p=0.01, Mann-Whitney U test). Multivariate analysis showed that 20-gauge PPV was a significant factor behind the increase in postoperative IL-8 (OR 5.21, 95% CI (1.46 to 18.55), p=0.018), but surgical time and simultaneous cataract surgery were not significant factors. CONCLUSIONS: The increase in inflammatory cytokine IL-8 in tear was significantly less after 23-gauge PPV than 20-gauge PPV, indicating that transconjunctival PPV is less invasive from the viewpoint of the ocular surface molecular biology. This information may be important when deciding the method of PPV.

Stromal cell-derived factor-1 is essential for photoreceptor cell protection in retinal detachment.

Stromal cell-derived factor-1 (SDF-1) causes chemotaxis of CXCR4-expressing bone marrow-derived cells. SDF-1 is involved in the pathogenesis of various vascular diseases, including those of the eye. However, the role of SDF-1 in neuronal diseases is not completely understood. Here, we show higher SDF-1 levels in the vitreous humor of patients with retinal detachment (RD) compared with normal patients. SDF-1 correlated positively with the duration as well as the extent of RD. Furthermore, SDF-1 correlated significantly with levels of interleukin-6 and interleukin-8, but not with vascular endothelial growth factor. Western blot analysis results showed significant SDF-1 up-regulation in detached rat retinas compared with normal animals. Immunohistochemistry data showed that SDF-1 was co-localized with the glial cells of the detached retina. SDF-1 blockade with a neutralizing antibody increased photoreceptor cell loss and macrophage accumulation in the subretinal space. The retinal precursor cell line R28 expressed CXCR4. SDF-1 rescued serum starvation-induced apoptosis in R28 cells and enhanced their ability to participate in wound closure in a scratch assay. Our results indicate a surprising, protective role for SDF-1 in RD. This effect may be mediated directly or indirectly through other cell types.

Predictable signs of benign course of polypoidal choroidal vasculopathy: based upon the long-term observation of non-treated eyes.

PURPOSE: To find predictable signs of benign polypoidal choroidal vasculopathy (PCV). METHODS: Medical records of 13 eyes from 12 patients who were followed up for 5 years or longer without treatment among 258 consecutive patients with PCV were reviewed retrospectively. The main outcomes measured were best corrected visual acuity (BCVA) and fundus findings during the follow-up period. RESULTS: The average age at presentation was 68 years, and the average follow-up period after diagnosis was 80 months (range, 62-119 months). The initial mean logarithmic value of the minimal angle of resolution (logMAR) BCVA was 0.28 +/- 0.26, and the final mean logMAR BCVA was 0.62 +/- 0.72. The difference in the logMAR BCVA values between the two points was not statistically significant (p > 0.05). The trend of change from baseline at 2-year follow-up was consistent with those at 5-year follow-up in nine eyes. Fundus findings at the initial examination were classified into two patterns: (i) reddish-orange nodules and detachment of the retinal pigment epithelium with/without detachment of the neurosensory retina (nine eyes); (ii) reddish-orange nodules alone, or nodules and small subretinal haemorrhage (four eyes). In the eyes with the first pattern, clinical course and visual prognosis were variable. An absence of hard exudates could be a sign to maintain a benign clinical course or stable vision with this pattern. The eyes with the second pattern took a benign clinical course with stable vision. CONCLUSIONS: There is certainly a group of PCV eyes with a benign prognosis. Considering the huge cost and risk of current therapies, the initial ocular findings could be deciding factors that determine the necessity for further treatment.

B cell-derived vascular endothelial growth factor A promotes lymphangiogenesis and high endothelial venule expansion in lymph nodes.

Vascular endothelial growth factor A (VEGF-A) is a prominent growth factor for both angiogenesis and lymphangiogenesis. Recent studies have shown the importance of VEGF-A in enhancing the growth of lymphatic endothelial cells in lymph nodes (LNs) and the migration of dendritic cells into LNs. VEGF-A is produced in inflamed tissues and/or in draining LNs, where B cells are a possible source of this growth factor. To study the effect of B cell-derived VEGF-A, we created transgenic mice (CD19(Cre)/hVEGF-A(fl)) that express human VEGF-A specifically in B cells. We found that the human VEGF-A produced by B cells not only induced lymphangiogenesis in LNs, but also induced the expansion of LNs and the development of high endothelial venules. Contrary to our expectation, we observed a significant decrease in the Ag-specific Ab production postimmunization with OVA and in the proinflammatory cytokine production postinoculation with LPS in these mice. Our findings suggest immunomodulatory effects of VEGF-A: B cell-derived VEGF-A promotes both lymphangiogenesis and angiogenesis within LNs, but then suppresses certain aspects of the ensuing immune responses.

Vitrectomy for exudative age-related macular degeneration with vitreous hemorrhage.

PURPOSE: The purpose of this study was to study the effect of pars plana vitrectomy (PPV) for age-related macular degeneration with vitreous hemorrhage on choroidal neo-vascularization (CNV). METHODS: A retrospective interventional case series in which 92 eyes with age-related macular degeneration with vitreous hemorrhage that received PPV were studied. Among them, 60 eyes without pre- or posttreatment other than PPV were selected. Choroidal neovascularization was expressed as the incidence of bleeding 6 months before and after PPV. The status of CNV after PPV was compared and classified as worsened, remained, regressed, disappeared, or unclassified. The influence of posterior vitreous detachment was examined. RESULTS: The incidence of bleeding was reduced dramatically after PPV (1.11 +/- 0.44 in preoperative 6 months vs. 0.03 +/- 0.18 in postoperative 6 months, P < 0.0001). The status of CNV improved in most cases; 40 of 54 classifiable eyes (74.1%) were categorized as "regressed" or "disappeared." Postoperative visual acuity was significantly better than preoperative visual acuity (P < 0.0001). The status of CNV subsided more in those eyes without posterior vitreous detachment than in those with posterior vitreous detachment (odds ratio, 1.02; 95% confidence interval, -0.01-2.08; P = 0.054). CONCLUSION: The activity of CNV was reduced after PPV in eyes with age-related macular degeneration with vitreous hemorrhage. Visual acuity significantly improved, with only rare severe complications. The involvement of vitreomacular traction in the patho-physiology of CNV in age-related macular degeneration is possible.

HMGB1: A new marker for estimation of the postmortem interval.

Estimation of the postmortem interval (PMI) is one of the most important tasks in forensic medicine. Numerous methods have been proposed for the determination of the time since death by chemical means. High mobility group box-1 (HMGB1), a nonhistone DNA-binding protein is released by eukaryotic cells upon necrosis. Postmortem serum levels of HMGB1 of 90 male Wistar rats stored at 4, 14 and 24°C since death were measured by enzyme-linked immunosorbent assay. The serum HMGB1 level showed a time-dependent increase up to seven days at 4°C. At 14°C, the HMGB1 level peaked at day 3, decreased at day 4, and then plateaued. At 24°C, the HMGB1 level peaked at day 2, decreased at day 3, and then plateaued. Our findings suggest that HMGB1 is related to the PMI in rats.

Edaravone attenuates cerebral ischemic injury by suppressing aquaporin-4.

Aquaporin-4 (AQP4) plays a role in the generation of post-ischemic edema. Pharmacological modulation of AQP4 function may thus provide a novel therapeutic strategy for the treatment of stroke, tumor-associated edema, epilepsy, traumatic brain injury, and other disorders of the central nervous system (CNS) associated with altered brain water balance. Edaravone, a free radical scavenger, is used for the treatment of acute ischemic stroke (AIS) in Japan. In this study, edaravone significantly reduced the infarct area and improved the neurological deficit scores at 24h after reperfusion in a rat transient focal ischemia model. Furthermore, edaravone markedly reduced AQP4 immunoreactivity and protein levels in the cerebral infarct area. In light of observations that edaravone specifically inhibited AQP4 in a rat transient focal ischemia model, we propose that edaravone might reduce cerebral edema through the inhibition of AQP4 expression following cerebral infarction.

Sonothrombolysis for intraocular fibrin formation in an animal model.

Vascular diseases such as diabetic retinopathy or retinal arterial occlusion are always associated with retinal and/or choroidal vasculopathy and intravascular thrombosis is commonly found. The ultrasound (US) therapy is a recently developed technique to accelerate fibrinolysis and it is being applied to some clinical fields. The present study was to observe the effects of extraocular US exposure on intraocular fibrin, which is a deteriorating factor in various ocular diseases. Tubes containing human blood (2 mL) in the following groups were irradiated with US; US alone, US with tissue plasminogen activator (tPA), tPA alone, and saline (control). Fibrinolysis was quantified by measuring D-dimer after 2h. In rat eyes, intracameral fibrin (fibrin formation in the anterior chamber of the eye) was induced by YAG-laser-induced iris bleeding. Then, eyes in the following groups were irradiated with US; US alone, subconjunctival tPA alone, US and subconjunctival tPA, control. Intracameral fibrin was scored on day 3 (3+ maximum to 0). The temperatures of rat eyes were measured by infrared thermography. Histologic evaluation was also performed. D-dimer was increased by US with statistical significance (p <0.05) or tPA (p <0.01). D-dimer in US with tPA group was significantly higher than either US alone or tPA alone group (p <0.01). In rat eyes, the average intracameral fibrin score on day 3 was 1.4 in control group and 1.2 in subconjunctival tPA alone group; however, it decreased significantly in the US alone group (0.75; p <0.05, vs. control), US and subconjunctival tPA group (0.71; p <0.01, vs. control). The temperature was less than 34 degrees C after US exposure. No histologic damage was observed. US irradiation from outside accelerated intracameral fibrinolysis without causing apparent tissue damage. This noninvasive method might have therapeutic value for intraocular fibrin.

Vitreous mediators after intravitreal bevacizumab or triamcinolone acetonide in eyes with proliferative diabetic retinopathy.

PURPOSE: To evaluate vitreous vascular endothelial growth factor (VEGF), stromal cell-derived factor 1alpha (SDF-1alpha), interleukins (ILs), and tumor necrosis factor-alpha (TNF-alpha) after intravitreal bevacizumab or triamcinolone acetonide (TA) in eyes with proliferative diabetic retinopathy (PDR). DESIGN: Interventional, consecutive, retrospective, comparative study with a historical control. PARTICIPANTS: Forty-seven eyes of 47 patients affected by active PDR were investigated. Bevacizumab (1.25 mg; 19 eyes; bevacizumab group) or TA (4 mg; 10 eyes; TA group) was injected into the vitreous cavity as preoperative adjunctive therapy 7 days before vitrectomy. Eighteen eyes without injection served as controls (control group). METHODS: The vitreous samples were obtained at vitrectomy and were analyzed for concentrations of total protein, VEGF, SDF-1alpha, IL-1beta, IL-6, IL-8, IL-10, IL-12p70, and TNF-alpha. MAIN OUTCOME MEASURES: Vitreous concentrations of VEGF, SDF-1alpha, ILs, and TNF-alpha were compared among bevacizumab, TA, and control groups. RESULTS: Vitreous concentrations of VEGF and SDF-1alpha were lower in bevacizumab and TA groups compared with the control group. The median VEGF level was 0 pg/ml (range, 0-79.2 pg/ml) in the bevacizumab group, 343.5 pg/ml (range, 0-1683.3 pg/ml) in the TA group, and 1202.5 pg/ml (range, 76-4213.9 pg/ml) in the control group. The median SDF-1alpha level was 149.2 pg/ml (range, 0-519.4 pg/ml) in the bevacizumab group, 87.5 pg/ml (range, 0-252.5 pg/ml) in the TA group, and 245.7 pg/ml (range, 0-856.8 pg/ml) in the control group. The differences in both vitreous VEGF and SDF-1alpha concentrations among 3 groups were statistically significant (P<0.001 and P = 0.010, respectively). The eyes with intravitreal bevacizumab demonstrated the lowest vitreous level of VEGF, and the level was statistically significant compared with the eyes with intravitreal TA and control eyes (P<0.001 and P<0.001, respectively). The control eyes had the highest vitreous level of SDF-1alpha, and the level was statistically significant compared with the eyes with intravitreal bevacizumab and TA (P = 0.015 and P = 0.009, respectively). There was no significant difference regarding ILs and TNF-alpha. CONCLUSIONS: Intravitreal injection of bevacizumab potentially diminishes not only VEGF but also SDF-1alpha. These findings suggest that intravitreal bevacizumab may influence intraocular mediators beyond VEGF. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Minocycline attenuates both OGD-induced HMGB1 release and HMGB1-induced cell death in ischemic neuronal injury in PC12 cells.

High mobility group box-1 (HMGB1), a non-histone DNA-binding protein, is massively released into the extracellular space from neuronal cells after ischemic insult and exacerbates brain tissue damage in rats. Minocycline is a semisynthetic second-generation tetracycline antibiotic which has recently been shown to be a promising neuroprotective agent. In this study, we found that minocycline inhibited HMGB1 release in oxygen-glucose deprivation (OGD)-treated PC12 cells and triggered the activation of p38mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinases (ERK1/2). The ERK kinase (MEK)1/2 inhibitor U-0126 and p38MAPK inhibitor SB203580 blocked HMGB1 release in response to OGD. Furthermore, HMGB1 triggered cell death in a dose-dependent fashion. Minocycline significantly rescued HMGB1-induced cell death in a dose-dependent manner. In light of recent observations as well as the good safety profile of minocycline in humans, we propose that minocycline might play a potent neuroprotective role through the inhibition of HMGB1-induced neuronal cell death in cerebral infarction.

The free radical scavenger edaravone rescues rats from cerebral infarction by attenuating the release of high-mobility group box-1 in neuronal cells.

Edaravone, a potent free radical scavenger, is clinically used for the treatment of cerebral infarction in Japan. Here, we examined the effects of edaravone on the dynamics of high-mobility group box-1 (HMGB1), which is a key mediator of ischemic-induced brain damage, during a 48-h postischemia/reperfusion period in rats and in oxygen-glucose-deprived (OGD) PC12 cells. HMGB1 immunoreactivity was observed in both the cytoplasm and the periphery of cells in the cerebral infarction area 2 h after reperfusion. Intravenous administration of 3 and 6 mg/kg edaravone significantly inhibited nuclear translocation and HMGB1 release in the penumbra area and caused a 26.5 +/- 10.4 and 43.8 +/- 0.5% reduction, respectively, of the total infarct area at 24 h after reperfusion. Moreover, edaravone also decreased plasma HMGB1 levels. In vitro, edaravone dose-dependently (1-10 microM) suppressed OGD- and H(2)O(2)-induced HMGB1 release in PC12 cells. Furthermore, edaravone (3-30 microM) blocked HMGB1-triggered apoptosis in PC12 cells. Our findings suggest a novel neuroprotective mechanism for edaravone that abrogates the release of HMGB1.