Retinal Histological Changes Research Articles

Retinal glial cell response in a neurodevelopmental disease such as the Dravet syndrome

Dravet syndrome (DS) is an epileptic encephalopathy caused by mutations in the Scn1a gene encoding the α1 subunit of the Nav1.1 sodium channel, which is associated with recurrent and generalized seizures, highly resistant to antiepileptic drugs, which is related to cognitive impairment, developmental delays, hyperactivity, dysautonomia, attention deficit, language impairment, autistic features, even leading to death. GABAergic interneurons are particularly affected, where the Nav1.1 channel is mainly located and the consequent imbalance between excitation and inhibition triggers brain hyperexcitability and seizures, altering the correct functioning of neural networks involved in cognitive processes.Glial reactivity and neuroinflammatory processes have been shown to be present in childhood epileptic processes and therefore cytokines may exert a neuromodulatory role directly impacting neuronal excitability. Indeed, astrocytes have been documented to be involved in the pathogenesis of epilepsy, regulating neurotransmitter and ion concentrations.In our experimental model of DS (Syn‐Cre/Scn1aWT/A1783V), inflammatory events in the prefrontal cortex and dentate gyrus of the hippocampus expressed as an increase in GFAP (astrocyte marker) and Iba‐1 (microglia marker) immunoreactivity have been observed, in addition to morphological changes compatible with activated astrocytes and microglia.The retina is a projection of the brain, and in several pathologies affecting the central nervous system, such as neurodegenerative diseases, correlation between changes occurring in the brain and those occurring in the retina has been shown. Therefore, the analyse of the changes that may occur in retinal glial cells (astrocytes and microglia) in an experimental model of DS, and the correlation with the alterations observed in some specific cerebral areas of these animals could be good predictive and prognostic biomarkers for future treatment.This study analyzes the retinal histological changes in glial cells (microglia and astrocytes), retinal ganglion cells (RGCs) and GABAergic amacrine cells in an experimental model of DS (Syn‐Cre/Scn1aWT/A1783V) compared to a control group at postnatal day (PND) 25. Retinal whole‐mounts were labelled with anti‐GFAP, anti‐Iba‐1, anti‐Brn3a and anti‐GAD65/67. Signs of microglial and astroglial activation and the number of Brn3a+ and GAD65+ 67+ cells were quantified. We found retinal activation of astroglial and microglial cells but not death of RGCs and GABAergic amacrine cells. These changes are similar to those found at the level of the hippocampus in the same experimental model in PND25, indicating a relationship between brain and retinal changes in DS. This suggests that the retina could serve as a possible biomarker in DS.

Retinal Tissue Shows Glial Changes in a Dravet Syndrome Knock-in Mouse Model.

Dravet syndrome (DS) is an epileptic encephalopathy caused by mutations in the Scn1a gene encoding the α1 subunit of the Nav1.1 sodium channel, which is associated with recurrent and generalized seizures, even leading to death. In experimental models of DS, histological alterations have been found in the brain; however, the retina is a projection of the brain and there are no studies that analyze the possible histological changes that may occur in the disease. This study analyzes the retinal histological changes in glial cells (microglia and astrocytes), retinal ganglion cells (RGCs) and GABAergic amacrine cells in an experimental model of DS (Syn-Cre/Scn1aWT/A1783V) compared to a control group at postnatal day (PND) 25. Retinal whole-mounts were labeled with anti-GFAP, anti-Iba-1, anti-Brn3a and anti-GAD65/67. Signs of microglial and astroglial activation, and the number of Brn3a+ and GAD65+67+ cells were quantified. We found retinal activation of astroglial and microglial cells but not death of RGCs and GABAergic amacrine cells. These changes are similar to those found at the level of the hippocampus in the same experimental model in PND25, indicating a relationship between brain and retinal changes in DS. This suggests that the retina could serve as a possible biomarker in DS.

Thioredoxin-1 Ameliorates Oxygen-Induced Retinopathy in Newborn Mice through Modulation of Proinflammatory and Angiogenic Factors.

Oxygen-induced retinopathy (OIR) is an animal model for retinopathy of prematurity, which is a leading cause of blindness in children. Thioredoxin-1 (TRX) is a small redox protein that has cytoprotective and anti-inflammatory properties in response to oxidative stress. The purpose of this study was to determine the effect of TRX on OIR in newborn mice. From postnatal day 7, C57BL/6 wild type (WT) and TRX transgenic (TRX-Tg) mice were exposed to either 21% or 75% oxygen for 5 days. Avascular and neovascular regions of the retinas were investigated using fluorescence immunostaining. Fluorescein isothiocyanate-dextran and Hoechst staining were used to measure retinal vascular leakage. mRNA expression levels of proinflammatory and angiogenic factors were analyzed using quantitative polymerase chain reaction. Retinal histological changes were detected using immunohistochemistry. In room air, the WT mice developed well-organized retinas. In contrast, exposing WT newborn mice to hyperoxia hampered retinal development, increasing the retinal avascular and neovascular areas. After hyperoxia exposure, TRX-Tg mice had enhanced retinal avascularization compared with WT mice. TRX-Tg mice had lower retinal neovascularization and retinal permeability during recovery from hyperoxia compared with WT mice. In the early stages after hyperoxia exposure, VEGF-A and CXCL-2 expression levels decreased, while IL-6 expression levels increased in WT newborn mice. Conversely, no differences in gene expressions were observed in the TRX-Tg mouse retina. IGF-1 and Angpt1 levels did not decrease during recovery from hyperoxia in TRX-Tg newborn mice. As a result, overexpression of TRX improves OIR in newborn mice by modulating proinflammatory and angiogenic factors.

Yiqi Tongluo Fang could preventive and delayed development and formation of diabetic retinopathy through antioxidant and anti-inflammatory effects

BackgroundYiqi Tongluo Fang (YQTLF) is an effective prescription for the treatment of diabetic retinopathy (DR), but its mechanism of action remains unclear. MethodThe content of YQTLF was determined using liquid and gas chromatography-mass spectrometry (LC-MS and GC-MS, respectively). Twenty-five Sprague Dawley (SD) rats were randomly selected as the normal control group. One hundred SD streptozotocin-induced diabetes (type 1) rats were randomly divided into diabetic control, diabetic+insulin+ calcium dobesilate (CaD), and diabetic+insulin+ YQTLF groups, with 25 rats in each group. Bodyweight level was measured every 2 weeks. After 12 weeks of gavage, the glucose levels, lipids, oxidative stress, inflammation, retinal histopathology, and the blood-retinal barrier were assessed in each group. The p38 MAPK pathway was changed to explore its internal mechanism. The measurement data were expressed as mean ± standard deviation, and different statistical methods were used according to a normal distribution, square error, or not. ResultsA total of 1024 valid peaks were identified in YQTLF using GC-MS. YQTLF significantly lowered the fasting blood glucose levels in diabetic rats. YQTLF early inhibited changes in retinal histology, capillaries, cells, and tight junction proteins (such as ZO-1, occludin, claudin-5, and VE-cadherin) before the formation and development of DR. These findings correlated with the alleviation of glucolipid metabolism, inflammation, and oxidative stress. The lncRNA MALAT1 and the PRC 2/p38 MAPK-related pathway, such as the expression of EZH2, SUZ12, EED, p38 MAPK, MMP-9, and VEGFR, were also correlated. ConclusionWe have demonstrated the molecular and cellular mechanisms underlying the preventive and delayed development and formation of DR. YQTLF prevents changes in dyslipidemia, retinal histology, capillaries, cells, and tight junction proteins. These protective effects appear to be linked to its antioxidant and anti-inflammatory effects, which prevent the activation of intracellular signaling pathways, such as the lncRNA MALAT1 and PRC 2/p38 MAPK-related pathway.

The role of hesperidin in ameliorating retinal changes in rats with experimentally induced type 1 diabetes mellitus and the active role of vascular endothelial growth factor and glial fibrillary acidic protein.

Patients with type 1 diabetes mellitus (T1DM) are vulnerable to developing diabetic retinopathy even under insulin therapy. Thus, this study was designed to evaluate the efficacy of hesperidin and insulin in rats with T1DM compared with insulin alone in improving diabetic retinal changes. Eighty rats were divided into four equal groups: group I, control rats without diabetes; group II, untreated rats with diabetes; group III, rats with diabetes treated daily with subcutaneous (SC) doses of long-acting insulin; and group IV, a rat with diabetes in which hesperidin was orally administered with SC insulin. The animals were assessed histologically, morphometrically, and biochemically. In group II, the thickness of all retinal layers decreased histologically. Ultrastructurally, degenerated retinal neurons and congested blood vessels were observed. Immunostaining detected elevated gene expression of advanced glycation end products. Gene expression of vascular endothelial growth factor, and glial fibrillary acidic protein were elevated. In this study, hesperidin supplementation with insulin significantly improved the retinal histological changes, supported by morphometric findings, compared with insulin alone. Moreover, treatment with hesperidin significantly reduced malondialdehyde and elevated serum antioxidant markers, including superoxide dismutase and catalase; furthermore, glutathione peroxidase decreased. Hesperidin might be an effective supplement for improving diabetic retinal complications occurring even with insulin treatment.

Study of retinal neurodegeneration and maculopathy in diabetic Meriones shawi: A particular animal model with human-like macula.

The purpose of this work was to evaluate a potentially useful animal model, Meriones shawi (M.sh)-developing metabolic X syndrome, diabetes and possessing a visual streak similar to human macula-in the study of diabetic retinopathy and diabetic macular edema (DME). Type 2 diabetes (T2D) was induced by high fat diet administration in M.sh. Body weights, blood glucose levels were monitored throughout the study. Diabetic retinal histopathology was evaluated 3 and 7 months after diabetes induction. Retinal thickness was measured, retinal cell types were labeled by immunohistochemistry and the number of stained elements were quantified. Apoptosis was determined with TUNEL assay. T2D induced progressive changes in retinal histology. A significant decrease of retinal thickness and glial reactivity was observed without an increase in apoptosis rate. Photoreceptor outer segment degeneration was evident, with a significant decrease in the number of all cones and M-cone subtype, but-surprisingly-an increase in S-cones. Damage of the pigment epithelium was also confirmed. A decrease in the number and labeling intensity of parvalbumin- and calretinin-positive amacrine cells and a loss of ganglion cells was detected. Other cell types showed no evident alterations. No DME-like condition was noticed even after 7 months. M.sh could be a useful model to study the evolution of diabetic retinal pathology and to identify the role of hypertension and dyslipidemia in the development of the reported alterations. Longer follow up would be needed to evaluate the potential use of the visual streak in modeling human macular diseases.

Transplantation of IL-10-transfected endothelial progenitor cells improves retinal vascular repair via suppressing inflammation in diabetic rats.

We aimed to evaluate the effect of IL-10 gene transfection on endothelial progenitor cells (EPCs) under inflammatory conditions, and explore the therapeutic potential of IL-10-transfected EPC transplantation on nonproliferative diabetic retinopathy (NPDR). Lentivirus vectors encoding IL-10 were constructed and introduced into EPCs isolated from rat bone marrow. After exposure to recombinant rat TNF-α, abilities of nontransfected EPCs (non-EPCs) and EPCs transfected with normal control lentivirus (EPCs-GFP) or IL-10 expressing lentivirus (EPCs-IL-10-GFP) were assessed, including migration, adhesion, and tube formation. IL-10 production by EPCs-IL-10-GFP was determined by ELISA. Following 12weeks after establishment of diabetes, diabetic rats were randomly injected with non-EPCs, EPCs-GFP, or EPCs-IL-10-GFP via tail vein. Expression of inflammatory factors and factors associated with nuclear factor-kappa B (NF-kB) signal pathway, retinal histological analysis, and retinal vascular permeability were assessed 2weeks after transplantation. The detrimental effects of TNF-ɑ on the abilities of EPCs were significantly attenuated in EPCs-IL-10-GFP compared with non-EPCs and EPCs-GFP. The concentration of IL-10 in the EPCs-IL-10-GFP group was significantly higher than the non-EPCs and EPCs-GFP groups. Additionally, transplantation of EPCs-IL-10-GFP significantly inhibited inflammatory factors expression and activation of NF-kB signal pathway, improved retinal histological changes, and attenuated retinal vascular permeability. In conclusion, transplantation of IL-10-transfected EPCs significantly improved EPCs-mediated retinal vascular repair and subsequently suppressed NPDR progression. This was associated with inflammation suppression, at least partly via inhibiting the NF-kB signal pathway. Transplantation of IL-10-transfected EPCs may be a new strategy for treatment of NPDR.

Safety evaluation of poly(lactic-co-glycolic acid)/poly(lactic-acid) microspheres through intravitreal injection in rabbits.

Poly(lactic-co-glycolic acid) (PLGA) and/or poly(lactic-acid) (PLA) microspheres are important drug delivery systems. This study investigated eye biocompatibility and safety of PLGA/PLA microspheres through intravitreal injection in rabbits. Normal New Zealand rabbits were randomly selected and received intravitreal administration of different doses (low, medium, or high) of PLGA/PLA microspheres and erythropoietin-loaded PLGA/PLA microspheres. The animals were clinically examined and sacrificed at 1, 2, 4, 8, and 12 weeks postadministration, and retinal tissues were prepared for analysis. Retinal reactions to the microspheres were evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick end staining and glial fibrillary acidic protein immunohistochemistry. Retinal structure changes were assessed by hematoxylin and eosin staining and transmission electron microscopy. Finally, retinal function influences were explored by the electroretinography test. Terminal deoxynucleotidyl transferase-mediated dUTP nick end staining revealed no apoptotic cells in the injected retinas; immunohistochemistry did not detect any increased glial fibrillary acidic protein expression. Hematoxylin and eosin staining and transmission electron microscopy revealed no micro- or ultrastructure changes in the retinas at different time points postintravitreal injection. The electroretinography test showed no significant influence of scotopic or photopic amplitudes. The results demonstrated that PLGA/PLA microspheres did not cause retinal histological changes or functional damage and were biocompatible and safe enough for intravitreal injection in rabbits for controlled drug delivery.

Diosmin Protects Rat Retina from Ischemia/Reperfusion Injury

Diosmin, a natural flavone glycoside, possesses antioxidant activity and has been used to alleviate ischemia/reperfusion (I/R) injury. The aim of this study was to clarify whether the administration of diosmin has a protective effect against I/R injury induced using the high intraocular pressure (IOP) model in rat retina, and to determine the possible antioxidant mechanisms involved. Retinal I/R injury was induced in the rats by elevating the IOP to 110 mmHg for 60 min. Diosmin (100 mg/kg) or vehicle solution was administered intragastrically 30 min before the onset of ischemia and then daily after I/R injury until the animals were sacrificed. The levels of malondialdehyde (MDA) and the activities of total-superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in the retinal tissues were determined 24 h after I/R injury. At 7 days post-I/R injury, electroretinograms (ERGs) were recorded, and the density of surviving retinal ganglion cells (RGCs) was estimated by counting retrograde tracer-labeled cells in whole-mounted retinas. Retinal histological changes were also examined and quantified using light microscopy. Diosmin significantly decreased the MDA levels and increased the activities of T-SOD, GSH-Px, and CAT in the retina of rats compared with the ischemia group (P<0.05), and suppressed the I/R-induced reduction in the a- and b-wave amplitudes of the ERG (P<0.05). The thickness of the entire retina, inner nuclear layer, inner plexiform layer, and outer retinal layer and the number of cells in the ganglion cell layer were significantly less after I/R injury (P<0.05), and diosmin remarkably ameliorated these changes on retinal morphology. Diosmin also attenuated the I/R-induced loss of RGCs of the rat retina (P<0.05). Diosmin protected the retina from I/R injury, possibly via a mechanism involving the regulation of oxidative parameters.

Valproic Acid Regulates Antioxidant Enzymes and Prevents Ischemia/Reperfusion Injury in the Rat Retina

Purposes: To investigate whether valproic acid (VPA) has a neuroprotective effect against ischemia/reperfusion (I/R) injury in the rat retina, and to elucidate the potential antioxidant mechanisms involved.Methods: Adult male Wistar rats were randomly divided into four groups: sham (group A), sham plus VPA (group B), I/R plus vehicle (group C), and I/R plus VPA (group D). Retinal I/R injury was produced by inducing an exceedingly high intraocular pressure (IOP). Prior to insult, VPA was administered subcutaneously (300 mg/kg twice daily) for 7 days, after which the animal was sacrificed. Levels of retinal malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were determined. Protein expressions of retinal poly(ADP-ribose) (PAR) and nitrotyrosine (NT) were analyzed by Western blotting 24 h after injury. Apoptosis of retinal cells was evaluated 24 h after I/R injury by immunofluorescence of activated caspase-3 in histological sections of retina. Seven days after reperfusion, electroretinography (ERG) was performed, and retinal histological changes were examined by light microscopy.Results: Following ischemia, the thickness of the entire retina, including the inner nuclear layer (INL) and inner plexiform layer (IPL), as well as the number of cells in the ganglion cell layer (GCL) were significantly greater in group D than in group C (p < 0.05). VPA suppressed I/R-induced reductions in ERG a- and b-wave amplitudes (p < 0.05). VPA attenuated I/R-induced activation of caspase-3 in ganglion cells and INL cells (p < 0.001). VPA significantly decreased MDA levels and increased activities of SOD, GSH-Px, and CAT in group D (p < 0.05). VPA attenuated activation of PAR and accumulation of NT in the retina after I/R (p < 0.01).Conclusions: VPA protects the retina from I/R injury by enhancing anti-oxidative effects and inhibiting apoptosis of retinal cells.

Gucy2f zebrafish knockdown – a model for Gucy2d-related leber congenital amaurosis

Mutations in retinal-specific guanylate cyclase (Gucy2d) are associated with Leber congenital amaurosis-1 (LCA1). Zebrafish offer unique advantages relative to rodents, including their excellent color vision, precocious retinal development, robust visual testing strategies, low cost, relatively easy transgenesis and shortened experimental times. In this study we will demonstrate the feasibility of using gene-targeting in the zebrafish as a model for the photoreceptor-specific GUCY2D-related LCA1, by reporting the visual phenotype and retinal histology resulting from Gucy2f knockdown. Gucy2f zebrafish LCA-orthologous cDNA was identified and isolated by PCR amplification. Its expression pattern was determined by whole-mount in-situ hybridization and its function was studied by gene knockdown using two different morpholino-modified oligos (MO), one that blocks translation of Gucy2f and one that blocks splicing of Gucy2f. Visual function was assessed with an optomotor assay on 6-days-post-fertilization larvae, and by analyzing changes in retinal histology. Gucy2f knockdown resulted in significantly lower vision as measured by the optomotor response compared with uninjected and control MO-injected zebrafish larvae. Histological changes in the Gucy2f-knockdown larvae included loss and shortening of cone and rod outer segments. A zebrafish model of Gucy2f-related LCA1 displays early visual dysfunction and photoreceptor layer dystrophy. This study serves as proof of concept for the use of zebrafish as a simple, inexpensive model with excellent vision on which further study of LCA-related genes is possible.

Retinal safety of a new fluoroquinolone, pradofloxacin, in cats: assessment with electroretinography

To investigate the safety of a new fluoroquinolone, pradofloxacin, on the cat retina using electroretinogram. Ganzfeld ERGs were recorded in 40 cats treated orally for 23 days in 4 groups: CTRL (n = 9): placebo-vehicle; PRADO30 (n = 10): pradofloxacin 30 mg/kg/day; PRADO50 (n = 14): pradofloxacin 50 mg/kg/day; and ENRO30 (n = 7): enrofloxacin at toxic doses of 30 mg/kg/day. ERG was performed before treatment and once weekly during the treatment period. An extended ISCEV protocol with addition of 8 steps of increasing luminance in dark adapted condition was carried out to assess: V (max) (saturated scotopic b-wave amplitude) and k (luminance inducing V (max)/2). OCT and retinal histological changes were also investigated. Pradofloxacin showed no effects in respect to rod b-wave, V (max), k and maximum scotopic a-wave (P > 0.05). Oscillatory potentials, cone ERG and flicker were also unaltered (P > 0.05). Rod b-wave was undetectable after treatment in ENRO30 group, V (max) was reduced to 10.5% of the baseline (P < 0.05), accompanied by an increase of k by 1 log cd s/m(2) (P < 0.05). Oscillatory potentials, cone b-wave amplitude and 30 Hz flicker amplitude were reduced to 8.3%, 58.9% and 37.4% of the baseline, respectively (P < 0.05). Effects were also seen in OCT and retinal histology starting within one week after the start of treatment and thereafter remaining stable. Pradofloxacin at 6 and 10 times the recommended doses was shown to have no retinal toxic effects in cats, neither on rod or cone function with ERG.

High infusion pressure in conjunction with vitreous surgery alters the morphology and function of the retina of rabbits

To investigate the effects of high infusion pressure in conjunction with pars plana vitrectomy (PPV) on retinal morphology and function in rabbits. Pars plana vitrectomy was performed under urethane (0.8 mg/kg) anaesthesia in the right eye of albino rabbits following phacoemulsification and aspiration (PEA). The left eyes were not touched. After PEA, the animals were divided into two groups. In six eyes, intraocular pressure (IOP) was increased to 80 mmHg for 30 mins (high-pressure group) and in five eyes IOP was maintained at 40 mmHg for 30 mins (low-pressure group). The IOPs were regulated by the height of the bottle of balanced salt solution (BSS) and monitored with a pressure transducer. After the pressure elevation, vitreous fluid was collected to measure the glutamate concentration. Then, PPV was performed for 15 mins in both groups under an infusion pressure of 40 mmHg. In five additional rabbits, PEA alone was performed in the right eye, and vitreous fluid was collected (PEA group). Functional alterations were assessed by recording visual evoked potentials (VEPs) and electroretinograms (ERGs). Ten days after the IOP changes, the animals were killed with intravenous pentobarbital sodium and the eyes were prepared for histological analysis. Damage to retinal ganglion cells (RGCs) was quantified by counting the number of cells in the ganglion cell layer (GCL). The contralateral eyes in the high-pressure group served as controls (n = 6). The mean implicit time (IT) of the VEPs in the high-pressure group was significantly longer than that before the IOP elevation, by 114-124% (p < 0.05, paired t-test), and also than that of control eyes (p < 0.05, anova followed by t-test). No significant changes in the VEPs were detected in either the low-pressure group or the PEA group. There were significantly fewer cells in the GCL in the high-pressure group (24.7/mm) than in the control animals (41.4/mm; p < 0.05, Dunnett's test). The number of cells in the GCL in the low-pressure and PEA groups did not significantly differ to that in the controls. The amplitudes of the ERG a- and b-waves were not significantly changed (p > 0.05, paired t-test). These results suggest that high infusion pressure in conjunction with PPV leads to morphological and functional changes in the retina. The absence of ERG changes and presence of VEP changes suggest that these changes were due to damage to RGCs, which supports the morphological observations.

Biodegradable implants for ocular delivery of anti-inflammatory drug

The treatment of vitreoretinal diseases is limited, and there are currently new drug delivery approaches being reported for increasing drug bioavailability. Intraocular implants can release drugs directly into the posterior segment of the eye and maintain long-term vitreous concentrations of drugs within the therapeutic range. The objective of this pilot study was to investigate the in vitro and in vivo short-term safety and pharmacokinetic behavior of a dexamethasone implant that is inserted into the vitreous using a new technique that does not require surgery. The results showed that the developed device prolonged the release of dexamethasone within therapeutic levels, both in vitro and in vivo. Furthermore, the implant was not associated with retinal histological changes or elevated intraocular pressure in normal rabbits’ eyes. Controlled animal studies are underway to test the efficacy of the implant in diseased eyes.

Pharmacokinetic and Toxicity Investigations of a New Intraocular Lens with a Dexamethasone Drug Delivery System: A Pilot Study

Aim: To investigate the short-term safety and pharmacokinetic behavior of a new intraocular lens containing a dexamethasone drug delivery system (IOL-DDS) in rabbit eyes. Methods: A modified polymethylmethacrylate IOL containing a biodegradable dexamethasone DDS was implanted into the posterior chamber of the right eyes of 9 New Zealand white rabbits. Serial slitlamp and indirect ophthalmoscopic examinations (including grading of intraocular inflammation) were performed. After 3, 6 and 9 days, the rabbits were euthanized and the globes were removed for histological examination and for determination of dexamethasone levels in the aqueous humor and in the vitreous. Analysis of dexamethasone concentrations was performed by ELISA. Results: Therapeutic concentrations of dexamethasone were detectable in the aqueous and vitreous of the study eyes throughout the 9-day period in all tested animals. The mean aqueous dexamethasone concentration (ng/ml, ±SD) was 1,015.42 (±43.05), 970.11 (±32.47) and 757.58 (±30.19) and the mean vitreous concentration (ng/ml, ±SD) was 399.82 (±38.05), 287.38 (±34.47) and 268.15 (±32.00) at 3, 6 and 9 days after the surgical procedure, respectively. No corneal or retinal histological changes were observed during the study period. Conclusion: The IOL-DDS is effective in delivering therapeutic concentrations of dexamethasone to the aqueous and vitreous, without acute damage to the cornea and retina. Further controlled studies in the same animal model are under way to determine the potential value of this lens in the prevention and treatment of inflammation following cataract surgery.

An experimental study on dissolving and detaching vitreoretinal interface with enzymes

To investigate the safety dosage of intravitreal injection of Plasmin, Hyaluronidase and their combination and to evaluate their efficacy in the formation of posterior vitreous detachment (PVD) through animal experiments. Eight rabbits each were randomly assigned to one of six groups and one eye of each rabbit were experimental eye, the other is control eye. The position of drug injection was in posterior 1/3 of vitreous before papillary. The drug-injection groups were assigned as following: the first group-Hyaluronidase 20 IU (0.1 ml BSS), the second group-Hyaluronidase 30 IU (0.1 ml BSS), the third group-Plasmin 1 IU (0.1 ml BSS), the forth group-Plasmin 2 IU (0.1 ml BSS), the fifth group-Plasmin 3 IU (0.1 ml BSS), the sixth group- Hyaluronidase 20 IU (0.05 ml BSS) combined with Plasmin 1 IU (0.05 ml BSS); the control eyes were injected with BSS (0.1 ml) at the same position. The experimental rabbits were followed for 2 weeks, and in this period all eyes were examined with slit-lamp biomicroscopy, +90 D preset lens, indirect ophthalmoscope, electroretinography, B-scan, and optical coherence tomography. After two weeks, the animals were killed and the eyes enucleated. Scan electron and light microscopy were performed. Hyaluronidase concentration of 20 IU, Plasmin concentration of 1 IU and their combination didn't produce inflammatory response and retinal toxicity in the experimental eyes. 30 IU Hyaluronidase and 2 IU Plasmin both caused mild inflammatory response in the vitreous without retinal histological changes, while the latter caused a reversible decrease in the ERG b-wave. 3 IU of Plasmin caused severe inflammatory response with retinal histological and electroretinographic changes. In safety dosage neither Hyaluronidase nor Plasmin alone were successful in producing completely posterior vitreous detachment (PVD), but the latter can produce partial PVD. The combination of Plasmin (1 IU) and Hyaluronidase (20 IU) was effective on producing completely PVD without retinal toxicity. 20 IU of Hyaluronidase and 1 IU of Plasmin are safe to retina and others structures in the eyes without any toxicity. Posterior vitreous injection of Hyaluronidase (20 IU) combined with Plasmin (1 IU) can produce complete posterior vitreous detachment.