Inflammatory and angiogenic protein detection in the human vitreous : cytometric bead assay

Introduction. To evaluate clinical feasibility and reproducibility of cytometric bead assay (CBA) in nondiluted vitreous samples of patients with age-related macular degeneration (ARMD), diabetic macular edema (DME), and central retinal vein occlusion (CRVO). Methods. Twelve patients from a single clinics day qualified for intravitreal injections (ARMD n = 6, DME n = 3, CRVO n = 3) and underwent a combination treatment including a single-site 23 gauge core vitrectomy which yielded a volume of 0.6 mL undiluted vitreous per patient. Interleukin-6 (IL-6), vascular endothelial growth factor isoform A (VEGF-A), and monocyte chemo-attractant protein-1 (MCP-1) were assessed directly from 0.3 mL at the same day (fresh samples). To assess the reproducibility 0.3 ml were frozen for 60 days at -80°, on which the CBA was repeated (frozen samples). Results. In the fresh samples IL-6 was highest in CRVO (median IL-6 55.8 pg/mL) > DME (50.6) > ARMD (3.1). Highest VEGF was measured in CRVO (447.4) > DME (3.9) > ARMD (2.0). MCP-1 was highest in CRVO (595.7) > AMD (530.8) > DME (178). The CBA reproducibility after frozen storage was examined to be most accurate for MCP1 (P = 0.91) > VEGF (P = 0.68) > IL-6 (P = 0.49). Conclusions. CBA is an innovative, fast determining, and reliable technology to analyze proteins in fluids, like the undiluted vitreous, which is important to better understand ocular pathophysiology and pharmacology. There is no influence of intermittent storage at -80° for the reproducibility of the CBA

Correlation from undiluted vitreous cytokines of untreated central retinal vein occlusion with spectral domain optical coherence tomography

Purpose: To correlate inflammatory and proangiogenic key cytokines from undiluted vitreous of treatment-naïve central retinal vein occlusion (CRVO) patients with SD-OCT parameters. Methods: Thirty-five patients (age 71.1 years, 24 phakic, 30 nonischemic) underwent intravitreal combination therapy, including a single-site 23-gauge core vitrectomy. Twenty-eight samples from patients with idiopathic, non-uveitis floaterectomy served as controls. Interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and vascular endothelial growth factor (VEGF-A) levels were correlated with the visual acuity (logMar), category of CRVO (ischemic or nonischemic) and morphologic parameters, such as central macular thickness-CMT, thickness of neurosensory retina-TNeuro, extent of serous retinal detachment-SRT and disintegrity of the IS/OS and others. Results: The mean IL-6 was 64.7pg/ml (SD ± 115.8), MCP-1 1015.7 ( ± 970.1), and VEGF-A 278.4 ( ± 512.8), which was significantly higher than the control IL-6 6.2 ± 3.4pg/ml (P=0.06), MCP-1 253.2 ± 73.5 (P<0.0000001) and VEGF-A 7.0 ± 4.9 (P<0.0006). All cytokines correlated highly with one another (correlation coefficient r=0.82 for IL-6 and MCP-1; r=0.68 for Il-6 and VEGF-A; r=0.64 for MCP-1 and VEGF-A). IL-6 correlated significantly with CMT, TRT, SRT, dIS/OS, and dELM. MCP-1 correlated significantly with SRT, dIS/OS, and dELM. VEGF-A correlated not with changes in SD-OCT, while it had a trend to be higher in the ischemic versus the nonischemic CRVO group (P=0.09). Conclusions: The inflammatory cytokines were more often correlated with morphologic changes assessed by SD-OCT, whereas VEGF-A did not correlate with CRVO-associated changes in SD-OCT. VEGF inhibition alone may not be sufficient in decreasing the inflammatory response in CRVO therapy

In Vivo Imaging of Intraocular Fluidics in Vitrectomized Swine Eyes Using a Digital Fluoroscopy System

properly cited. Purpose. To describe the characteristics of intraocular fluidics during cataract surgery in swine eyes with prior vitrectomy. Methods. We prepared three groups of enucleated swine eyes (nonvitrectomized, core, and totally vitrectomized). Irrigation and aspiration were performed (2.7 mm conventional sleeved phacosystem) using a balanced saline solution mixed with a water-soluble radiopaque contrast medium at 1 : 1 ratio. We imaged the eyes using a digital fluoroscopy system (DFS) during phacoemulsification and compared the characteristics of the intraocular fluid dynamics between the groups. Results. The anterior chamber depth (ACD) after the commencement of irrigation differed between groups (2.25 ± 0.06 mm; 2.33 ± 0.06 mm; 3.17 ± 0.11 mm), as well as the height of the fluid flowing from the anterior chamber into the posterior cavity that was identified by lifting up the iris to correct the infusion deviation syndrome (0.00 ± 0.00 mm; 0.41 ± 0.04 mm; 2.19 ± 0.35 mm). Conclusions. DFS demonstrated differences in fluid dynamics during phacoemulsification in swine eyes with or without prior vitrectomy. In completely vitrectomized eyes, the large ACD, which developed during phacoemulsification, could be reduced by lifting the iris and allowing the fluid to shift to the posterior cavity. Recognizing the differences in fluidics of vitrectomized eyes as compared to those of the nonvitrectomized eyes may reduce the frequency of intraoperative complications

Impact of Indocyanine Green Concentration, Exposure Time, and Degree of Dissolution in Creating Toxic Anterior Segment Syndrome: Evaluation in a Rabbit Model

Purpose. To investigate the role of indocyanine green (ICG) dye as a causative material of toxic anterior segment syndrome (TASS) in an experimental rabbit model. Method. Eight eyes of four rabbits were allocated to this study. Capsular staining was performed using ICG dye, after which the anterior chamber was irrigated with a balanced salt solution. The effects of different concentrations (control, 0.25, 0.5, and 1.0%), exposure times (10 and 60 seconds), and the degree of dissolution (differently vortexed) were investigated. The analysis involved anterior segment photography, ultrasound pachymetry, prostaglandin assay (PGE2 Parameter Assay, R&D systems, Inc.), and scanning electron microscopy of each iris. Result. There was no reaction in the control eye. A higher aqueous level of PGE2 and more severe inflammatory reaction were observed in cases of eyes with higher concentration, longer exposure time, and poorly dissolved dye. Additionally, scanning electron microscopy revealed larger and coarser ICG particles. Conclusion. TASS occurrence may be associated with the concentration, exposure time, and degree of dissolution of ICG dye during cataract surgery

In Vivo Imaging of Intraocular Fluidics in Vitrectomized Swine Eyes Using a Digital Fluoroscopy System

Purpose. To describe the characteristics of intraocular fluidics during cataract surgery in swine eyes with prior vitrectomy. Methods. We prepared three groups of enucleated swine eyes (nonvitrectomized, core, and totally vitrectomized). Irrigation and aspiration were performed (2.7 mm conventional sleeved phacosystem) using a balanced saline solution mixed with a water-soluble radiopaque contrast medium at 1 : 1 ratio. We imaged the eyes using a digital fluoroscopy system (DFS) during phacoemulsification and compared the characteristics of the intraocular fluid dynamics between the groups. Results. The anterior chamber depth (ACD) after the commencement of irrigation differed between groups (2.25±0.06 mm; 2.33±0.06 mm; 3.17±0.11 mm), as well as the height of the fluid flowing from the anterior chamber into the posterior cavity that was identified by lifting up the iris to correct the infusion deviation syndrome (0.00±0.00 mm; 0.41±0.04 mm; 2.19±0.35 mm). Conclusions. DFS demonstrated differences in fluid dynamics during phacoemulsification in swine eyes with or without prior vitrectomy. In completely vitrectomized eyes, the large ACD, which developed during phacoemulsification, could be reduced by lifting the iris and allowing the fluid to shift to the posterior cavity. Recognizing the differences in fluidics of vitrectomized eyes as compared to those of the nonvitrectomized eyes may reduce the frequency of intraoperative complications

Profile of ocriplasmin and its potential in the treatment of vitreomacular adhesion

The recent approval by the US Food and Drug Administration of ocriplasmin for the treatment of symptomatic vitreomacular adhesion (VMA), often associated with vitreomacular traction (VMT) and macular hole (MH), has brought new attention to the field of pharmacologic vitreolysis. The need for an enzyme to split the vitreomacular interface, which is formed by a strong adhesive interaction between the posterior vitreous cortex and the internal limiting membrane, historically stems from pediatric eye surgery. This review summarizes the different anatomic classifications of posterior vitreous detachment or anomalous posterior vitreous detachment and puts these in the context of clinical pathologies commonly observed in clinical practice of the vitreoretinal specialist, such as MH, VMT, age-related macular degeneration, and diabetic macular edema. We revisit the outcome of the Phase II studies that indicated ocriplasmin was a safe and effective treatment for selected cases of symptomatic VMA and MH. Release of VMA at day 28 was achieved by 26.5% of patients in the ocriplasmin group versus 10.1% in the placebo group (P<0.001). Interestingly, for MHs, the numbers were more remarkable. Predictive factors for successful ocriplasmin treatment were identified for VMT (VMA diameter smaller than 1,500 µm) and MH (smaller than 250 µm). In comparison with the highly predictable outcome after vitrectomy, the general success rate of ocriplasmin not under clinical trial conditions has not fully met expectations and needs to be proven in real-world clinical settings. The ocriplasmin data will be compared in the future with observational data on spontaneous VMA release, will help retina specialists make more accurate predictions, and will improve outcome rates

Intravitreal sustained-release steroid implants for the treatment of macular edema following surgical removal of epiretinal membranes

Purpose: To evaluate the efficacy of intravitreal dexamethasone implant for the treatment of postoperative persistent cystoid macular edema (CME) following macular pucker surgery. Methods: In this multicenter study, we retrospectively reviewed the data of 37 patients (39 eyes) who had been treated with intravitreal dexamethasone implant (Ozurdex®) for persistent CME following macular pucker surgery. Main outcome measures were change in best-corrected visual acuity (BCVA) and central retinal thickness (CRT). Results: All eyes underwent spectral domain optical coherence tomography examination within 130 days after implantation. We observed a significant decrease in mean CRT from 519.9 to 392.9 µm (p < 0.0001). By this time, mean BCVA had improved from 0.60 to 0.43 logMAR (p = 0.003). Seventeen eyes (43.6%) required at least 1 repeat injection of dexamethasone. Of these, 8 (47%) eyes received a total number of 3 or more dexamethasone injections. Conclusion: Intravitreal dexamethasone implant injection is an effective treatment option for persistent CME following macular pucker surgery. Keywords: Central retinal thickness; Dexamethasone implant; Fluocinolone acetonide implant; Intravitreal steroid implants; Long-term outcome; Macular pucker surgery; Membrane peeling; Pars plana vitrectomy; Persistent macular edema

Clinical efficacy of simulated vitreoretinal surgery to prepare surgeons for the upcoming intervention in the operating room

Purpose: To evaluate the efficacy of the virtual reality training simulator Eyesi to prepare surgeons for performing pars plana vitrectomies and its potential to predict the surgeons’ performance. Methods: In a preparation phase, four participating vitreoretinal surgeons performed repeated simulator training with predefined tasks. If a surgeon was assigned to perform a vitrectomy for the management of complex retinal detachment after a surgical break of at least 60 hours it was randomly decided whether a warmup training on the simulator was required (n = 9) or not (n = 12). Performance at the simulator was measured using the built-in scoring metrics. The surgical performance was determined by two blinded observers who analyzed the video-recorded interventions. One of them repeated the analysis to check for intra-observer consistency. The surgical performance of the interventions with and without simulator training was compared. In addition, for the surgeries with simulator training, the simulator performance was compared to the performance in the operating room. Results: Comparing each surgeon’s performance with and without warmup trainingshowed a significant effect of warmup training onto the final outcome in the operating room. For the surgeries that were preceeded by the warmup procedure, the performance at the simulator was compared with the operating room performance. We found that there is a significant relation. The governing factor of low scores in the simulator were iatrogenic retinal holes, bleedings and lens damage. Surgeons who caused minor damage in the simulation also performed well in the operating room. Conclusions: Despite the large variation of conditions, the effect of a warmup training as well as a relation between the performance at the simulator and in the operating room was found with statistical significance. Simulator training is able to serve as a warmup to increase the average performance