[Effect of Selective Retina Therapy (SRT) on the Inflammatory Microenvironment of the Subretinal Space]. / Einfluss der Selektiven Retinatherapie (SRT) auf inflammatorische Zellmediatoren des subretinalen Raums.

PURPOSE: To investigate the effect of Selective Retina Therapy (SRT) on inflammatory key factors such as complement factor-C3 (CC3), tumor growth factor-beta2 (TGF-ß2), tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ). MATERIALS AND METHODS: Porcine RPE-Bruch's membrane-choroid explants were irradiated with two SRT laser systems, SRTYLF and SRTYAG (Nd : YLF laser, wave length 527 nm, pulse duration 1.7 µs and Nd : YAG laser, wave length 532 nm, pulse duration 2.4 - 3 µs). Laser irradiation was performed on a spot size of 200 × 200 µm, 30 pulses, with a repetition rate of 100 Hz, and a radiant exposure of 140 (threshold RPE death) and 180 mJ/cm2 per pulse (above threshold RPE death). Explants were cultivated in modified Ussing chambers and culture viability was assessed by calcein-AM cell staining. Secretion of inflammatory factors was analyzed by ELISA. Protein expression of tissue explants was assessed by Western blot. RESULTS: Regeneration of RPE was observed after 4 days. One day after SRT with 140 mJ/cm2 per pulse the secretion of basal CC3 decreased in ELISA. Following 180 mJ/cm2 radiant exposure, the level of IFN-γ decreased at day 4. CONCLUSION: SRT does not induce the release of the pro-inflammatory factors analyzed in this in-vitro study.

Long-term results of glaucoma drainage device surgery.

BACKGROUND: To evaluate long-term results of eyes with glaucoma drainage device (GDD). METHODS: We retrospectively reviewed medical records of all patients who underwent GDD placement at our institution between 2001 and 2014. A total of 110 eyes of 90 patients were studied. Glaucoma outcome was assessed by postoperative intraocular pressure (IOP), number of medications, and need for further glaucoma surgery. Surgical procedures before and during the study period, and their complications were evaluated. RESULTS: The mean follow-up was 78.3 ± 44.0 months. The mean preoperative intraocular pressure was 30.8 ± 6.9 mmHg with 3.5 ± 1.1 glaucoma medications. At last postoperative follow-up, the mean IOP decreased to 14.3 ± 5.4 mmHg with 1.6 ± 1.5 glaucoma medications. GDD implantation successfully controlled glaucoma in 86, 85, 81, 78, 79, 76 and 73% of eyes at 1, 2, 3, 4, 5, 7 and 10 years, respectively. At last follow-up IOP was successfully controlled in 67% of eyes. Clinical complications occurred in 56.4% of eyes during the follow-up period. CONCLUSIONS: A glaucoma drainage device can successfully control intractable glaucoma even after a very long period of time.

Open globe and penetrating eyelid injuries from fish hooks.

BACKGROUND: A few case reports have described accidental eye injuries caused by fish hooks. The severity of ocular injuries is dependent on the involved ocular structures. Severe ocular injuries due to fish hooks are rare. We describe open globe and penetrating eyelid injuries from fish hooks at the Baltic Sea. METHODS: Nine patients with traumatic ocular injuries caused by fish hooks were included. The following parameters were evaluated: severity of injury, best corrected visual acuity at admission and last follow-up, and surgical treatment. RESULTS: All nine patients were male. Age ranged between 7 and 51 years with a median of 13 years. Sixty-seven percent of the patients were children. Four of the nine patients were 9 years or younger. In 5 eyes (55%) the injury was limited to the eyelid. An open globe injury was found in 4 patients (45%). The mean follow-up was 16.7 ± 32.8 months. All patients required surgical treatment. The number of operations ranged from 1 to 3, with a mean of 1.4. At admission and last follow-up, patients with eyelid injuries showed a median best corrected visual acuity (BCVA) of logMAR 0.0. Patients with open globe injuries showed a median best corrected visual acuity of logMAR 1.5 at admission, and of logMAR 0.6 at last follow-up. CONCLUSIONS: Nearly half of the patients suffered severe penetrating injuries. Especially children misjudge the risk potential of fishing due to their lack of experience. Fishing glasses should be worn not only for UV protection, but also as injury prevention strategy.

Retinal volume change is a reliable OCT biomarker for disease activity in neovascular AMD.

PURPOSE: Current algorithms for automated computer interpretation of optical coherence tomography (OCT) imaging of patients suffering from neovascular age-related macular degeneration (AMD) mostly rely on fluid detection. However, fluid detection itself and correct interpretation of the fluid currently limits diagnostic accuracy. We therefore performed a detailed analysis of the requirements that would have to be met for fluid detection approaches. We further investigated if monitoring retinal volume would be a viable alternative to detect disease activity. METHODS: Retrospective analysis and manual grading of 764 OCT volume scans of 44 patients with exudative AMD treated with intravitreal anti-VEGF injections at a pro-re-nata (PRN) treatment regimen for at least 24 months. RESULTS: Detection of subretinal fluid (SRF) or intraretinal fluid (IRF) alone is not sufficient for disease detection. A combination of SRF and IRF can detect disease activity with a sensitivity of 98.6% and a specificity of 82%. With further characterization of IRF into exudative and degenerative cysts, specificity can be increased to 100%. However, correct characterization is currently not achieved by published fluid detection approaches. Change of macular retinal volume (MRV) can depict disease activity with sensitivity of 88.4% and specificity of 89.6%. Combination with the detection of SRF can further improve diagnostic accuracy to a specificity of 93.3% and sensitivity of 93.9% without relying on IRF or IRF characterization. CONCLUSION: Fluid detection without further characterization is not sufficient for AMD monitoring. Either further distinction between exudative and degenerative cysts is necessary, or other activity markers have to be taken into account. MRV offers good potential to fill this diagnostic gap and might become an important monitoring marker.

Long-term follow-up of lamellar macular holes and pseudoholes over at least 5 years.

PURPOSE: To assess morphological and functional changes of lamellar macular holes and pseudoholes with or without vitrectomy and membrane peeling with at least 5 years follow-up. METHODS: Retrospective study of 73 eyes with lamellar macular hole (LH, n = 28), macular pseudohole (PH, n = 31), and pseudohole with cleaved edges (cleavedPH, n = 14). Forty-six eyes were merely observed without vitreoretinal intervention (observation group), and 27 eyes underwent vitrectomy with membrane peeling (vitrectomy group). Outcome measures were best corrected visual acuity (BCVA) and morphological retinal parameters evaluated with optical coherence tomography (TD-OCT and SD-OCT). RESULTS: Mean follow-up was 8.3 years (5-12); mean age was 67 years (46-84). In the observation group, median BCVA (logMAR) at first exam was 0.2 (LH), 0.1 (PH), 0.2 (cleavedPH) and at last exam 0.3 (LH, p = 0.02), 0.2 (PH), 0.15 (cleavedPH). In the vitrectomy group, median BCVA at first exam was 0.4 (LH), 0.3 (PH), 0.25 (cleavedPH); before vitrectomy BCVA was 0.5 (LH), 0.35 (PH), 0.35 (cleavedPH); and at last exam BCVA increased to 0.3 (LH), 0.2 (PH, p < 0.05), 0.1 (cleavedPH, p < 0.05). At last exam, BCVA of LH was significantly worse compared to PH and cleavedPH. In the observation group, 6 of 29 eyes with PH or cleavedPH showed a spontaneous resolution of the epiretinal membrane with improvement of the foveal contour. Nine of 16 eyes with LH and 2/20 eyes with PH presented lamellar hole-associated epiretinal proliferation (LHEP) in SD-OCT. CONCLUSIONS: LH, PH, and cleavedPH are often stable over a very long time. LH tends to worse visual function compared to PH and cleavedPH. A spontaneous separation of epiretinal membranes in the long-term is not uncommon. Vitreoretinal intervention should be considered in cases with significant visual loss or functional and morphological progression.

Persistent Full-Thickness Idiopathic Macular Hole: Anatomical and Functional Outcome of Revitrectomy with Autologous Platelet Concentrate or Autologous Whole Blood.

PURPOSE: To evaluate long-term outcome after revitrectomy with autologous platelet concentrate (APC) or whole blood (WB) in persistent idiopathic macular hole (MH) after vitrectomy with internal limiting membrane (ILM) peeling. PROCEDURES: Retrospective study of 75 eyes with persistent MH after vitrectomy with ILM peeling and gas. All patients underwent revitrectomy with gas and APC (n = 61) or WB (n = 14). Main outcome measures were anatomical closure rate and postoperative best-corrected visual acuity (BCVA). RESULTS: Closure rate after revitrectomy was 85.2% (52/61) in the APC group and 7.1% (1/14) in the WB group. Median follow-up was 58 (range 3-147) months. Median BCVA (logMAR) in patients with finally closed MHs was 0.4 ± 0.3. Patients with defects of the ellipsoid zone had significantly worse postoperative BCVA. Morphological MH configuration (atrophic or elevated edges) did not correlate with final closure rate. CONCLUSIONS: Revitrectomy with APC and gas is a very effective treatment in persistent MH after vitrectomy with ILM peeling and gas.

Thinning of Inner Retinal Layers after Vitrectomy with Silicone Oil versus Gas Endotamponade in Eyes with Macula-Off Retinal Detachment.

PURPOSE: To evaluate retinal layer thickness with optical coherence tomography (OCT) in eyes with macula-off retinal detachment after silicone oil (SiO) or gas endotamponade. PROCEDURES: Cross-sectional study of 40 eyes with macula-off rhegmatogenous retinal detachment that underwent vitrectomy. 20 eyes received SiO tamponade and 20 matched eyes received gas. 33 healthy fellow eyes served as controls. Macular spectral domain OCT was performed with automated layer detection in the 5 inner subfields of the Early Treatment Diabetic Retinopathy Study (ETDRS) map. RESULTS: Comparing the SiO group with the gas group, the ganglion cell layer showed a significant thinning in all fields of the inner ring of the ETDRS map, the inner plexiform layer in the nasal, superior and temporal quadrants, and the outer plexiform layer in the nasal quadrant. CONCLUSIONS: Inner retinal layers in the fovea/parafovea were significantly thinner in the SiO group. Prospective studies are warranted to further elucidate possible retinal adverse effects of SiO tamponade.

Vision loss under silicone oil tamponade.

PURPOSE: We aimed to investigate frequency, time course and pathophysiology of vision loss in eyes with macula-on rhegmatogenous retinal detachment operated with vitrectomy and silicone oil tamponade. PATIENTS AND METHODS: Fifteen eyes of 15 patients who had been operated with 5,000 centistoke silicone oil between 2006 and 2014 were included in a retrospective case series. Examinations included logMAR best corrected visual acuity (BCVA), visual field testing (VF), spectral domain optical coherence tomography (OCT), electrophysiology, and fluorescein angiography. RESULTS: Vision loss was seen in eight (53 %) eyes of 15 patients with symptomatic central scotoma, which was confirmed by VF (5/6). Preoperative median BCVA of these patients was 0.15 (0.5 to 0), prior to oil removal 0.7 (1.0 to 0.5), and 6 weeks post oil removal 1.0 (1.5 to 0.2). BCVA recovered in five patients to a median of 0.15 (0.5 to 0.1), and it remained 1.0 in three (20 %) out of 15 eyes. OCT revealed significant thinning of the foveal and parafoveal combined nerve fiber, ganglion cell and inner plexiform layers in affected eyes (mean 58.3 µm +/-13, horizontal scan through fovea, 500 µm radius) compared to their healthy fellow eyes (mean 84.5 µm +/-12.3; p < 0.01, n = 6 patients, 12 eyes) and compared to eyes with no vision loss under silicone oil. CONCLUSIONS: We find persisting vision loss in three out of 15 patients treated for macula-on rhegmatogenous retinal detachment with silicone oil tamponade. Thinning of inner retinal layers possibly evoked by silicone oil tamponade might be a pathophysiological explanation for vision loss in these patients.

Basal and apical regulation of VEGF-A and placenta growth factor in the RPE/choroid and primary RPE.

PURPOSE: Members of the vascular endothelial growth factor (VEGF) family are strongly involved in pathological processes in the retina, such as age-related macular degeneration and diabetic retinopathy. Cells of the retinal pigment epithelium (RPE) constitutively secrete VEGF-A, and the secretion of placental growth factor (PlGF) has also been described. RPE cells are strongly polarized cells with different secretome at the apical and basal side. In this study, we evaluated the basal and apical regulation of VEGF-A and PlGF secretion in RPE/choroid explants and primary RPE cells. METHODS: RPE/choroid tissue explants were prepared from porcine eyes and cultivated in modified Ussing chambers, separating apical (RPE) and basal (choroid) supernatant. Primary RPE cells were also prepared from porcine eyes and cultivated on Transwell plates. Explants and cells were treated with inhibitors for VEGFR-2 (SU1498), p38 (SB203580), and the transcription factors nuclear factor-kappa B (NF-κB) and SP-1 (mithramycin), respectively. VEGF-A and PlGF content was evaluated with enzyme-linked immunosorbent assay (ELISA). In addition, western blots were performed. RESULTS: In the RPE/choroid, VEGF-A can initially be found on the apical and basal sides with significantly more pronounced secretion on the basal side. VEGF-A secretion is differentially regulated on the apical and basal sides, with the inhibition of SP-1 and NF-κB showing strong effects apically and basally after 24 h and 48 h, the inhibition of p38 displaying its effect mainly on the basal side with some effect apically after 48 h, and the inhibition of VEGFR-2 reducing the secretion of VEGF only on the apical side at 24 h and 48 h. In the RPE cell culture, similar effects were found, with inhibition of NF-κB or SP-1 displaying a strong decrease in VEGF-A on both sides, and p38 inhibition displaying only an inhibitory effect on the basal side. In contrast, an apical effect of VEGFR-2 inhibition was not found. However, the western blot experiments exhibited a significant decrease in the VEGF-A protein under SU1498 treatment. In the RPE/choroid organ cultures, PlGF was initially found mainly on the basal site with only minute amounts of PlGF found apically. NF-κB and SP-1 were strongly involved in PlGF regulation apically and basally, while VEGFR2 and to a lesser degree p38 displayed some regulation at the basal site. In the primary RPE cell culture, PlGF was not found on the apical or basal side. CONCLUSIONS: VEGF-A and PlGF were constitutively secreted and regulated by the RPE/choroid complex, with PlGF secreted mainly by the choroid. Although the transcription factors NF-κB and SP-1 were involved in apical and basal regulation of both growth factors, VEGFR-2 displayed a strong polarity, with regulation of apical VEGF-A and basal PlGF secretion.

Retinal Vascular Occlusion after COVID-19 Vaccination: More Coincidence than Causal Relationship? Data from a Retrospective Multicentre Study.

BACKGROUND: To investigate whether vaccination against SARS-CoV-2 is associated with the onset of retinal vascular occlusive disease (RVOD). METHODS: In this multicentre study, data from patients with central and branch retinal vein occlusion (CRVO and BRVO), central and branch retinal artery occlusion (CRAO and BRAO), and anterior ischaemic optic neuropathy (AION) were retrospectively collected during a 2-month index period (1 June-31 July 2021) according to a defined protocol. The relation to any previous vaccination was documented for the consecutive case series. Numbers of RVOD and COVID-19 vaccination were investigated in a case-by-case analysis. A case-control study using age- and sex-matched controls from the general population (study participants from the Gutenberg Health Study) and an adjusted conditional logistic regression analysis was conducted. RESULTS: Four hundred and twenty-one subjects presenting during the index period (61 days) were enrolled: one hundred and twenty-one patients with CRVO, seventy-five with BRVO, fifty-six with CRAO, sixty-five with BRAO, and one hundred and four with AION. Three hundred and thirty-two (78.9%) patients had been vaccinated before the onset of RVOD. The vaccines given were BNT162b2/BioNTech/Pfizer (n = 221), followed by ChadOx1/AstraZeneca (n = 57), mRNA-1273/Moderna (n = 21), and Ad26.COV2.S/Johnson & Johnson (n = 11; unknown n = 22). Our case-control analysis integrating population-based data from the GHS yielded no evidence of an increased risk after COVID-19 vaccination (OR = 0.93; 95% CI: 0.60-1.45, p = 0.75) in connection with a vaccination within a 4-week window. CONCLUSIONS: To date, there has been no evidence of any association between SARS-CoV-2 vaccination and a higher RVOD risk.

Influence of alcohol consumption on incidence and severity of open-globe eye injuries in adults.

AIM: To investigate the influence of alcohol consumption on the occurrence of open-globe injuries in adults. METHODS: A retrospective study was made of 100 consecutive patients (81 male, 19 female) with open-globe injuries. Of these patients, 18 exhibited alcohol intoxication (group Ai), and 82 exhibited no alcohol intoxication (group nAi). Investigated parameters were best-corrected visual acuity at day of admission and last examination (logMAR), type of injury according to BETT-classification, extraocular injuries, cause of injury, time and setting of injury, in relation to alcohol consumption and tested for statistical significance with Fisher's exact test or the Mann-Whitney U test, respectively. RESULTS: In group Ai, 83.3% of the patients were male, and in group nAi, 80.5%. Mean logMAR at day of admission was 1.06 ± 0.63 (20/250) in group Ai and 1.08 ± 0.59 (20/250) in group nAi. At last examination, mean logMAR in group Ai was 1.11 ± 0.59 (20/250), in group nAi 0.75 ± 0.60 (20/125). This difference was statistically significant (p = 0.02). In group Ai, significantly more ruptures according to BETT classification occurred (p = 0.05). In group Ai, significantly more additional extraocular injuries occurred compared to group nAi (38.9% versus 6.1%; p = 0.0009). In group Ai, the cause of injury was significantly more often glass (44.4% versus 2.4%; p = 0.0000), in group nAi the injury was more often directly or indirectly caused by tools (74.4% versus 33.3%; p = 0.001). In group Ai, the injury was significantly more often inflicted by others (50.0% versus 9.8%; p = 0.0003). The settings in which the injuries occurred were significantly more often the street in group Ai (44.4% versus 6.1%; p = 0.0002), in group nAi the garden or tool shed (31.7% versus 5.6%; p = 0.02) or the workplace (34.2 % versus 11.1 %; p = 0.04). In group Ai, the injuries occurred significantly more often at night (p = 0.0001) and on weekends (p = 0.0000). CONCLUSIONS: Open-globe eye injuries under alcohol intoxication are more often caused by a third party and have a worse prognosis. Open-globe injuries under alcohol intoxication occur in a different spatio-temporal setting and exhibit a more severe type of injury. Risk behavior combined with alcohol consumption therefore seems to be an independent factor for the incidence of open-globe eye injuries.

Safety testing of indocyanine green with different surgical light sources and the protective effect of optical filters.

PURPOSE: The purpose of this study was to assess the light-induced cytotoxicity of indocyanine green (ICG) using different light sources commonly used in macular surgery and to assess the effect of optical filters. METHODS: Primary cultures of porcine retinal pigment epithelium cells were incubated with 0.5 mg/mL ICG solution dissolved in 5% glucose and illuminated with a surgical light fiber for 3 or 15 minutes. Halogen, mercury vapor, xenon, and metal halide light sources were used. Cell viability was assessed using the MTT assay. Retinal pigment epithelium cells without illumination served as controls. The decomposition of ICG after illumination was analyzed by high-performance liquid chromatography. RESULTS: Illumination of retinal pigment epithelium cells with all light sources with or without previous incubation with ICG did not affect cell viability compared with controls. Cell viability was significantly reduced when the cells were not rinsed immediately after incubation. The cytotoxic effect was abolished by a 475-nm long-pass filter. The high-performance liquid chromatography analysis of the illuminated ICG solution identified six cytotoxic ICG decomposition products. CONCLUSION: Optical filters that narrow the emission spectrum of the light sources reduce the light-induced cytotoxicity of ICG. Optical filters applied in ICG-assisted macular surgery may reduce the risk of intraoperative cell damage.

[Vitreous body hemorrhage-How long can one wait?] / Glaskörperblutung ­ Wie lange kann man abwarten?

In the case of acute vitreous body hemorrhage, the ophthalmologist is faced with the question of whether spontaneous absorption can be awaited or whether a vitrectomy is necessary. In addition to the intensity of the bleeding, the underlying cause also plays an important role. A thorough medical history is therefore important. The most common causes in adults include posterior vitreous detachment, diabetic retinopathy, and retinal vascular occlusion. In children, retinal vasculitis, systemic diseases, and trauma (e.g. shaken baby syndrome) must be excluded. If there is no evidence of a retinal tear or detachment using ultrasound, it is usually possible to wait a few weeks under sonographic control to assess the spontaneous tendency to resorb. Overall, the trend is towards earlier surgical interventions due to improved vitrectomy techniques and the desire for early rehabilitation of vision.

Immobilization during anesthesia for vitrectomy using a laryngeal mask without neuromuscular blockade versus endotracheal intubation and neuromuscular blockade.

BACKGROUND: For intraocular surgery, most authors recommend general anesthesia including intubation and neuromuscular blockade to avoid complications by patient movements. However, anesthesia using a laryngeal mask and avoidance of muscle relaxants is common clinical practice. Purpose of this prospective observational study was to compare the incidence of eye movement and deviation of the eye axis during general anesthesia adjusted to minimal alveolar concentration (MAC) for pars plana-vitrectomy (PPV) using a laryngeal mask without neuromuscular blockade (LM) versus endotracheal intubation and neuromuscular blockade (INT). METHODS: The patients (N.=148) who underwent PPV for vitreoretinal disorders received MAC adjusted general anesthesia by volatile anesthetics. Seventy-four patients were subjected to LM and 74 to INT. In both groups the patient's lungs were mechanically ventilated without allowing spontaneous ventilation. Eye movements and upward deviations were judged and documented during surgery. Postoperatively quality of immobilization was assessed by a standardized questionnaire. RESULTS: Intraoperative movements were not observed in either group. Upward eye deviation was observed in nine (12%) patients in the LM group, but not in patients in the INT group (0%; P=0.003). The standardized questionnaire revealed no clinically relevant difference in quality of immobilization between groups, especially eye deviations did not lead to any relevant surgical complications or difficulties. CONCLUSIONS: For PPV, MAC adjusted balanced anesthesia using a laryngeal mask without neuromuscular blockade was associated with more, but clinically irrelevant upward eye deviations and may be an alternative to intubation with neuromuscular blockade. However, adequate depth of anesthesia must be assured to avoid unwanted injuries during surgery.

Autologous retinal pigment epithelium-choroid sheet transplantation in age related macular degeneration: morphological and functional results.

OBJECTIVE: To evaluate the outcome of autologous retinal pigment epithelium (RPE)-choroid sheet transplantation after removal of a subfoveal choroidal neovascularisation (CNV) in patients with age related macular degeneration (AMD). METHODS: RPE-choroid sheet transplantation was performed in 10 consecutive patients with exudative AMD (n = 9) or geographic atrophy (n = 1). After CNV extraction, an autologous RPE-choroid patch was translocated from the midperiphery under the macula. Follow-up was between 6 and 12 months. Visual acuity testing and microperimetry (Nidek-MP1) as well as autofluorescence, fluorescein and indocyanine green (ICG) angiography were performed and the data were analysed retrospectively. RESULTS: Visual acuity (logarithm of minimum angel of resolution) before operation ranged from 0.7 to 1.8 (mean 1.37) and after operation from 0.4 to 1.6 (mean 1.24). Visual acuity after operation improved in seven patients (by a mean of 0.26), remained stable in one patient and decreased in two patients. Microperimetry showed light sensitivity and fixation on the sheet in five cases. ICG angiography demonstrated perfusion through the RPE-choroid graft in nine patients. Postoperative complications included retinal detachment (n = 1) and epiretinal membrane formation (n = 2). The patient with geographic atrophy developed a CNV after surgery. CONCLUSIONS: Autologous RPE-choroid sheet transplantation is feasible and a comparatively safe procedure. Microperimetry showed fixation and light perception over the graft with a moderate increase in mean visual acuity.

Preoperative aqueous humour flare values do not predict proliferative vitreoretinopathy in patients with rhegmatogenous retinal detachment.

BACKGROUND/AIMS: Patients with rhegmatogenous retinal detachment (RRD) who develop postoperative proliferative vitreoretinopathy (PVR) have been found to have higher preoperative laser flare values than patients with RRD who do not develop this complication. Measurement of laser flare has therefore been proposed as an objective, rapid and non-invasive method for identifying high-risk patients. The purpose of our study was to validate the use of preoperative flare values as a predictor of PVR risk in two additional patient cohorts, and to confirm the sensitivity and specificity of this method for identifying high-risk patients. METHODS: We combined data from two independent prospective studies: centre 1 (120 patients) and centre 2 (194 patients). Preoperative aqueous humour flare was measured with a Kowa FM-500 Laser Flare Meter. PVR was defined as redetachment due to the formation of traction membranes that required reoperation within 6 months of initial surgery. Logistic regression and receiver operating characteristic analysis determined whether higher preoperative flare values were associated with an increased risk of postoperative PVR. RESULTS: PVR redetachment developed in 21/314 patients (6.7%). Median flare values differed significantly between centres, therefore analyses were done separately. Logistic regression showed a small but statistically significant increase in odds with increasing flare only for centre 2 (OR 1.014; p=0.005). Areas under the receiver operating characteristic showed low sensitivity and specificity: centre 1, 0.634 (95% CI 0.440 to 0.829) and centre 2, 0.731 (95% CI 0.598 to 0.865). CONCLUSIONS: Preoperative laser flare measurements are inaccurate in discriminating between those patients with RRD at high and low risk of developing PVR.

Compatibility of recombinant tissue plasminogen activator (rtPA) and aflibercept or ranibizumab coapplied for neovascular age-related macular degeneration with submacular haemorrhage.

BACKGROUND/AIMS: Subretinal coapplication of recombinant tissue plasminogen activator (rtPA) and vascular endothelial growth factor (VEGF)-antagonists is a new treatment option for age-related macular degeneration complicated by submacular haemorrhage. Here, we investigate the compatibility of rtPA and aflibercept or ranibizumab in vitro because intraoperatively, rtPA or rtPA-induced plasmin may cleave aflibercept or ranibizumab. METHODS: Aflibercept and ranibizumab, respectively, were incubated with rtPA or plasmin, separated in gel electrophoresis and stained with Coomassie or silver. The antiangiogenic activity of the VEGF-antagonists was quantified by VEGF-ELISA after incubation with the supernatant of primary porcine retinal pigment epithelium cell cultures. RESULTS: In electrophoresis, ranibizumab displayed no additional fragments when it was coapplied with rtPA or plasmin. Its VEGF-inhibiting efficacy remained unchanged in coapplication with rtPA with or without blood, or plasmin. rtPA did not cleave or functionally compromise aflibercept. When aflibercept was coapplied with plasmin, electrophoresis displayed additional bands in Coomassie (30 kDa, 27 kDa, 19 kDa, 15 kDa) and silver staining (31 kDa, 26 kDa, 21 kDa, 19 kDa, 15 kDa). While at a clinical dosage (800 µg/mL) VEGF was inhibited by aflibercept when coapplied with plasmin, at borderline concentrations (400 ng/mL) VEGF-binding ability of aflibercept was abolished. CONCLUSIONS: Ranibizumab is not cleaved or functionally compromised by rtPA or plasmin. Aflibercept is cleaved and its VEGF-binding ability is reduced when coapplied with plasmin. In clinical practice, rtPA and ranibizumab can be coapplied as a treatment for neovascular age-related macular degeneration with submacular haemorrhage while the antiangiogenic activity of aflibercept may be compromised when coapplied with rtPA in the presence of plasmin.

Effects of Cytokine Activation and Oxidative Stress on the Function of the Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells.

PURPOSE: In several retinal complications, such as age-dependent macular degeneration (AMD), oxidative stress is increased and cytokine level is elevated. These are shown to alter the activation and expression of matrix metalloproteinase (MMP) both in human primary and immortalized retinal pigment epithelial (RPE) cells. However, the effects on human embryonic stem cell (hESC)-derived RPE cells remain to be elucidated. METHODS: The mature hESC-RPE cells were exposed to inflammatory cytokines (IFN-γ or TNF-α) for 24 hours or oxidative stress (H2O2) for 1 hour. Effects on barrier properties were analyzed with transepithelial electrical resistance (TEER), the expression of MMP-1, MMP-2, MMP-3, MMP-9, collagen I, and collagen IV genes with quantitative RT-PCR, and the expression of MMP-1 and MMP-3 proteins with Western blot or ELISA, respectively. Also, activation and secretion of MMP-2 and -9 proteins were analyzed with zymography. RESULTS: In normal state, mature hESC-RPE cells expressed MMP-1, -2, -3, and -9 genes in low levels, respectively. Tumor necrosis factor-α increased MMP-1 and -2 gene expression, and H2O2 increased MMP-3 and -9 gene expression. Zymography revealed IFN-γ- and TNF-α-induced secretion of MMP-2 and high-molecular-weight species of MMP (HMW MMP), but H2O2 decreased their secretion. Furthermore, TNF-α and H2O2 significantly decreased barrier properties. CONCLUSIONS: Here, cytokines induced the MMP-1 and -2 gene and protein expression. Also, H2O2 induced MMP-3 and -9 gene expression, but not their protein secretion. These data propose that under oxidative stress and cytokine stimuli, mature hESC-RPE cells resemble their native counterpart in the human eye in regard to MMP secretion and expression and could be used to model retinal disorders involving alterations in MMP activity such as AMD, diabetic retinopathy, or proliferative vitreoretinopathy in vitro.

Dimethylfumarate is a potent inducer of apoptosis in human T cells.

Fumaric acid esters (FAE) have been used for the systemic treatment of psoriasis in Germany for almost 50 years. Recently, it has been shown that dimethylfumarate (DMF) as the main ingredient of the marketed FAE mixture is a potent inhibitor of the nuclear transcription factor NF-kappaB. DMF was also shown to induce apoptosis in various cells. Because T cells play a crucial role in psoriasis pathogenesis, we asked whether DMF and its main metabolite methylhydrogenfumarate (MHF) were able to induce apoptosis in these cells. Purified human T cells were treated with DMF and MHF (1-20 microg/mL) and stimulated with interleukin 2, anti-CD3 antibodies or both for 48 h, and apoptosis was subsequently determined by the expression of Apo2.7 as well as by terminal deoxynucleotide transferase nick end labeling. The expression of antiapoptotic protein Bcl-2 was simultaneously determined. The results showed a dose-and-time dependent up-regulation of Apo2.7 expression and DNA fragmentation by DMF preferable in stimulated T cells. MHF and the solvent dimethyl sulfoxide were without effect. DMF, but not MHF, led to a concentration-dependent decrease of Bcl-2 expression in interleukin-2-stimulated T cells. The data provide evidence that the effect of FAE treatment of psoriasis may at least in part be due to induction of apoptosis in activated T cells.