Pathological high intraocular pressure induces glial cell reactive proliferation contributing to neuroinflammation of the blood-retinal barrier via the NOX2/ET-1 axis-controlled ERK1/2 pathway.

BACKGROUND: NADPH oxidase (NOX), a primary source of endothelial reactive oxygen species (ROS), is considered a key event in disrupting the integrity of the blood-retinal barrier. Abnormalities in neurovascular-coupled immune signaling herald the loss of ganglion cells in glaucoma. Persistent microglia-driven inflammation and cellular innate immune system dysregulation often lead to deteriorating retinal degeneration. However, the crosstalk between NOX and the retinal immune environment remains unresolved. Here, we investigate the interaction between oxidative stress and neuroinflammation in glaucoma by genetic defects of NOX2 or its regulation via gp91ds-tat. METHODS: Ex vivo cultures of retinal explants from wildtype C57BL/6J and Nox2 -/- mice were subjected to normal and high hydrostatic pressure (Pressure 60 mmHg) for 24 h. In vivo, high intraocular pressure (H-IOP) was induced in C57BL/6J mice for two weeks. Both Pressure 60 mmHg retinas and H-IOP mice were treated with either gp91ds-tat (a NOX2-specific inhibitor). Proteomic analysis was performed on control, H-IOP, and treatment with gp91ds-tat retinas to identify differentially expressed proteins (DEPs). The study also evaluated various glaucoma phenotypes, including IOP, retinal ganglion cell (RGC) functionality, and optic nerve (ON) degeneration. The superoxide (O2-) levels assay, blood-retinal barrier degradation, gliosis, neuroinflammation, enzyme-linked immunosorbent assay (ELISA), western blotting, and quantitative PCR were performed in this study. RESULTS: We found that NOX2-specific deletion or activity inhibition effectively attenuated retinal oxidative stress, immune dysregulation, the internal blood-retinal barrier (iBRB) injury, neurovascular unit (NVU) dysfunction, RGC loss, and ON axonal degeneration following H-IOP. Mechanistically, we unveiled for the first time that NOX2-dependent ROS-driven pro-inflammatory signaling, where NOX2/ROS induces endothelium-derived endothelin-1 (ET-1) overexpression, which activates the ERK1/2 signaling pathway and mediates the shift of microglia activation to a pro-inflammatory M1 phenotype, thereby triggering a neuroinflammatory outburst. CONCLUSIONS: Collectively, we demonstrate for the first time that NOX2 deletion or gp91ds-tat inhibition attenuates iBRB injury and NVU dysfunction to rescue glaucomatous RGC loss and ON axon degeneration, which is associated with inhibition of the ET-1/ERK1/2-transduced shift of microglial cell activation toward a pro-inflammatory M1 phenotype, highlighting NOX2 as a potential target for novel neuroprotective therapies in glaucoma management.

The Impact of Aging on the Function of Retinal Ganglion Cells.

Aging is a major risk factor for retinal neurodegenerative diseases. Aged mammalian retinal ganglion cells (RGCs) lack the ability to regenerate axons after injury. Rodent models suggest that older age increases the vulnerability of RGCs to injury and impairs RGC function as well as their functional recovery. Molecular changes - including decreased circulating levels of brain-derived neurotrophic factor (BDNF) - might contribute to impaired RGC dendritic extension during aging. Moreover, age-related mitochondrial dysfunction plays a major role in aging processes, as it leads to reduced adenosine triphosphate and increased generation of reactive oxygen species. Autophagy activity is necessary for the maintenance of cellular homeostasis and decreases with aging in the central nervous system. During aging, vascular insufficiency may lead to impaired oxygen and nutrient supply to RGCs. Microglial cells undergo morphological changes and functional impairment with aging, which might compromise retinal homeostasis and promote an inflammatory environment. Addressing these age-related changes by means of a low-energy diet, exercise, and neurotrophic factors might prevent age-related functional impairment of RGCs. This review focuses on the current understanding of aging RGCs and key players modulating those underlying mechanisms.

The Association between Vascular Abnormalities and Glaucoma-What Comes First?

Glaucoma is a leading cause of irreversible blindness worldwide. While intraocular pressure (IOP) presents a major risk factor, the underlying pathophysiology still remains largely unclear. The correlation between vascular abnormalities and glaucoma has been deliberated for decades. Evidence for a role played by vascular factors in the pathogenesis of glaucomatous neurodegeneration has already been postulated. In addition, the fact that glaucoma causes both structural and functional changes to retinal blood vessels has been described. This review aims to investigate the published evidence concerning the relationship between vascular abnormalities and glaucoma, and to provide an overview of the "chicken or egg" dilemma in glaucoma. In this study, several biomarkers of glaucoma progression from a vascular perspective, including endothelin-1 (ET-1), nitric oxide, vascular endothelial growth factor (VEGF), and matrix metalloproteinases (MMPs), were identified and subsequently assessed for their potential as pharmacological intervention targets.

Glaucoma as a Neurodegenerative and Inflammatory Disease.

Glaucoma is a neurodegenerative disease that leads to irreversible loss of vision through degeneration of the retinal ganglia cells (RGCs). Glaucoma is one of the most frequent causes of blindness in the world. Intraocular pressure is the main risk factor for the occurrence and development of this disease. Treatment is largely based on reducing internal optical pressure. However, some patients may deteriorate or become blind, despite normal or reduced internal optical pressure. The pathophysiological details are still unclear. Neuroinflammatory processes are also apparently an additional cause. In principle, innate or local responses of the adaptive immune system can be distinguished. The reaction of the innate immune system, particularly the local microglial cells, has long been studied. The macroglia with the astrocytes and Müller cells and their homeostatic effects have also long been known. On the other hand, it has long been thought that the retina with its RGZs was inert to adaptive immunological reactions - due to the function of the blood brain barrier. However, this system may be disturbed by antigen presentation, leading to a reaction of the adaptive immune system, with B cell and T cell responses. In this context, the key proteins are presumably heat shock proteins. We now know that neuroinflammation is important in glaucoma, as in other neurodegenerative diseases. It is important to increase our understanding of these phenomena. In this review article, we present our current knowledge of the role of the micro- and macroglia, the adaptive immune system, and the heat shock proteins.

[Normal Tension Glaucoma]. / Normaldruckglaukome.

Normal tension glaucoma (NTG) is a subvariant of primary open angle glaucoma (POAG) or a separate entity. NTG is defined as chronic open angle glaucoma without elevated intraocular pressure or normal intraocular pressure. Normal intraocular pressure is between 10 and 21 mmHg and is defined as two times the standard deviation of the mean intraocular pressure in the normal population. In addition to the absence of elevated intraocular pressure, all the classic symptoms of glaucoma are otherwise present. These include a conspicuous glaucomatous optic disc excavation, nerve fibre bundle defects, and corresponding visual field defects. Papillary rim haemorrhages are frequently found. The visual field defects in NDG are usually more central compared to POAG and are therefore described as more disturbing by the patient. The anterior chamber angle is open and there are no other changes suggestive of secondary glaucoma (pigment dispersion, pseudo-exfoliation). The exact pathophysiology of NDG is not well understood. Pathophysiologically, circulatory disturbances in the sense of arterial hypo- as well as hypertension may play an essential role or at least increase the susceptibility of the optic nerve to intraocular pressure fluctuations as well as blood pressure dips. Therefore, this requires not only a purely ophthalmologic but also interdisciplinary treatment of the patient with confirmed NDG. The primary goal of treatment is the reduction of intraocular pressure, which can stop the disease. This article gives an overview of epidemiology, aetiology, clinical findings and therapies.

Chronic social defeat stress causes retinal vascular dysfunction.

PURPOSE: The roles of vascular dysfunction and chronic stress have been extensively discussed in the pathophysiology of glaucoma. Our aim was to test whether chronic stress causes retinal vascular dysfunction and therewith induces retinal ganglion cells (RGCs) loss. METHODS: Twelve mice underwent chronic social defeat (CSD) stress, while 12 mice received control treatment only. Intraocular pressure (IOP) was measured with a rebound tonometer. Blood plasma corticosterone concentration and adrenal gland weight were used to assess stress levels. Brn-3a staining in retinas and PPD staining in optic nerve cross sections were conducted to assess the survival of RGCs and axons respectively. The ET-1 and α-SMA levels were determined in retina. Retinal vascular autoregulation, functional response to various vasoactive agents and vascular mechanics were measured using video microscopy. RESULTS: No significant difference in IOP levels was observed during and after CSD between CSD mice and controls. CSD stress caused hypercortisolemia 2 days post-CSD. However, increased corticosterone levels went back to normal 8 months after CSD. CSD-exposed mice developed adrenal hyperplasia 3 days post-CSD, which was normalized by 8 months. RGC and axon survival were similar between CSD mice and controls. However, CSD stress caused irreversible, impaired autoregulation and vascular dysfunction of retinal arterioles in CSD mice. In addition, impaired maximal dilator capacity of retinal arterioles was observed 8 months post-CSD rather than 3 days post-CSD. Remarkably, ET-1 levels were increased 3 days post-CSD while α-SMA levels were decreased 8 months post-CSD. CONCLUSIONS: We found that CSD stress does not cause IOP elevation, nor loss of RGCs and their axons. However, it strikingly causes irreversible impaired autoregulation and endothelial function in murine retinal arterioles. In addition, CSD changed vascular mechanics on a long-term basis. Increased ET-1 levels and loss of pericytes in retina vessels may involve in this process.

Energy Metabolism in the Inner Retina in Health and Glaucoma.

Glaucoma, the leading cause of irreversible blindness, is a heterogeneous group of diseases characterized by progressive loss of retinal ganglion cells (RGCs) and their axons and leads to visual loss and blindness. Risk factors for the onset and progression of glaucoma include systemic and ocular factors such as older age, lower ocular perfusion pressure, and intraocular pressure (IOP). Early signs of RGC damage comprise impairment of axonal transport, downregulation of specific genes and metabolic changes. The brain is often cited to be the highest energy-demanding tissue of the human body. The retina is estimated to have equally high demands. RGCs are particularly active in metabolism and vulnerable to energy insufficiency. Understanding the energy metabolism of the inner retina, especially of the RGCs, is pivotal for understanding glaucoma's pathophysiology. Here we review the key contributors to the high energy demands in the retina and the distinguishing features of energy metabolism of the inner retina. The major features of glaucoma include progressive cell death of retinal ganglions and optic nerve damage. Therefore, this review focuses on the energetic budget of the retinal ganglion cells, optic nerve and the relevant cells that surround them.

Hydrogen Sulfide: Novel Endogenous and Exogenous Modulator of Oxidative Stress in Retinal Degeneration Diseases.

Oxidative stress (OS) damage can cause significant injury to cells, which is related to the occurrence and development of many diseases. This pathological process is considered to be the first step to trigger the death of outer retinal neurons, which is related to the pathology of retinal degenerative diseases. Hydrogen sulfide (H2S) has recently received widespread attention as a physiological signal molecule and gas neuromodulator and plays an important role in regulating OS in eyes. In this article, we reviewed the OS responses and regulatory mechanisms of H2S and its donors as endogenous and exogenous regulators in retinal degenerative diseases. Understanding the relevant mechanisms will help to identify the therapeutic potential of H2S in retinal degenerative diseases.

[Endothelial Cell Reaction to Elevated Hydrostatic Pressure and Oxidative Stress in Vitro]. / Reaktion der Endothelzellen auf kurzzeitig physiologisch erhöhte hydrostatische Drücke und oxidativen Stress in vitro.

INTRODUCTION: Endothelial dysfunction has become a strongly discussed factor regarding glaucoma pathogenesis. In addition to peripapillary bleedings as signs of vascular damage, there is a definite correlation between glaucoma and vascular dysregulation syndrome. The aim of this study was to evaluate endothelial cell reaction to moderately elevated hydrostatic pressure and oxidative stress in vitro. METHODS: In vitro, primarily dissociated brain microvascular endothelial cells (BMECs) were exposed to moderately elevated hydrostatic pressure (60 and 120 mmHg) in a special pressure chamber. Additionally, cells primarily exposed to pressure, and cells not exposed to pressure, were incubated with low amounts of H2O2. A live/dead assay was performed to evaluate cell viability. Immunohistochemical staining against actin was used for morphological evaluation. RESULTS: Neither 60 nor 120 mmHg of elevated pressure had a viability changing effect on primary endothelial cells. Secondary, no big morphological changes could be discovered. However, against a low concentration of oxidative stress, BMECs showed high vulnerability. A difference in reaction to cells stressed with high pressure before could not be shown. CONCLUSION: Direct effects, in terms of higher vulnerability or morphological changes of moderately elevated high pressure on endothelial cells, could not be shown. However, the reaction to low amounts of oxidative stress indicates the involvement of endothelial cells in the pathogenesis of glaucoma and the special role of oxidative stress when referring to endothelial dysfunction in glaucomatous disease.

[Morphological and Quantitative Changes in Retinal and Optic Nerve Vessels in Experimental Glaucoma Model with Elevated IOP for 7 Weeks]. / Erhöhter Augeninnendruck für 7 Wochen induziert lokale Gefäßveränderungen im experimentellen Glaukommodell in vivo.

INTRODUCTION: Glaucoma is characterised by progressive loss of retinal ganglion cells and axons. Experimental research has concentrated on understanding the pathophysiological mechanisms involved in glaucomatous damage. It is still a matter of debate whether neurons or capillaries are primarily damaged by elevated intraocular pressure (IOP). The aim of this study was to detect IOP-induced vascular changes in the vessels of the optic nerve head and the main vessels of the retina in vivo. METHODS: Experimental glaucoma was induced in adult Sprague Dawley rats by cauterisation of three episcleral veins of the left eye (n = 3). In vivo, retinal vessel calibre was measured manually using a peripapillary scan with SD-OCT (Heidelberg Engineering) at baseline and after seven weeks of IOP elevation. The animals were then sacrificed and the optic nerve was fixed with 30% glutaraldehyde and cross-sections stained with paraphenylene diamine to mark the vessels. Contralateral eyes served as controls. Pictures were taken and number of vessels, vessel calibre and area were calculated and compared. RESULTS: IOP was significantly elevated (p < 0.001). In optic nerve cross sections, the number of capillaries did not differ significantly between animals with elevated IOP and controls. However, vessel calibre and area were significantly reduced (p < 0.001) in glaucomatous optic nerves. The calibre of the retinal vessels was significantly lowered - by 9.22% (p = 0.021). CONCLUSION: Retinal arterioles and optic nerve capillaries respond sensitively to abnormal pressure elevation in vivo, showing high and early vulnerability. The vascular responses may influence secondary neuronal responses, which culminate in the death of ganglion cells and blindness, as occurs in clinical glaucoma.

Proteomic profiling reveals crucial retinal protein alterations in the early phase of an experimental glaucoma model.

PURPOSE: Clinical glaucoma is difficult to assess in terms of molecular pathophysiology, prompting studies in experimental models of glaucoma. The purpose of this study was to investigate quantitative changes in retinal protein expression at the onset of experimental glaucoma in rats. Analyzing the proteome provides a suitable tool to decipher the pathophysiological processes in glaucomatous degeneration. METHODS: Thermic cauterization of episcleral veins was utilized to elevate the intraocular pressure in Sprague Dawley rats. Morphological changes were surveyed on a cellular level with a staining of Brn3a-positive cells. The retinal nerve fiber layer was investigated using optical coherence tomography (OCT, Heidelberg Engineering) and the optic nerve was analyzed by an axonal grading system. Mass spectrometry-featured quantitative proteomics and immunohistochemical staining was used to identify specifically altered proteins in the course of intraocular pressure elevation and initial neurodegeneration. Proteomic data were further analyzed with Ingenuity Pathway Analysis and Cytoscape to analyze further molecular associations. RESULTS: The intraocular pressure rose significantly (p < 0.001) for the follow-up period of 3 weeks after which animals were sacrificed. Eyes exposed to an elevated intraocular pressure showed an initial decrease of retinal ganglion cells, retinal nerve fiber layer (p < 0.05) and an impairment of the optic nerve (p < 0.01). Mass spectrometry led to the identification and quantification of 931 retinal proteins, whereas 32 were considerably altered. Bioinformatics-assisted clustering revealed that a majority of these proteins are functionally associated with cell differentiation, apoptosis and stress response. The creation of an interactive protein network showed that numerous altered proteins are connected regarding their cellular function. Protein kinase b, mitogen-activated protein kinase 1 and the NF-κB complex seem to be essential molecules in this context. CONCLUSIONS: In conclusion, these results provide further lines of evidence that substantial molecular changes occur at the onset of the disease, identifying potential key players, which might be useful as biomarkers for diagnostics and development of medical treatment in the future.

The Small Heat Shock Protein α-Crystallin B Shows Neuroprotective Properties in a Glaucoma Animal Model.

Glaucoma is a neurodegenerative disease that leads to irreversible retinal ganglion cell (RGC) loss and is one of the main causes of blindness worldwide. The pathogenesis of glaucoma remains unclear, and novel approaches for neuroprotective treatments are urgently needed. Previous studies have revealed significant down-regulation of α-crystallin B as an initial reaction to elevated intraocular pressure (IOP), followed by a clear but delayed up-regulation, suggesting that this small heat-shock protein plays a pathophysiological role in the disease. This study analyzed the neuroprotective effect of α-crystallin B in an experimental animal model of glaucoma. Significant IOP elevation induced by episcleral vein cauterization resulted in a considerable impairment of the RGCs and the retinal nerve fiber layer. An intravitreal injection of α-crystallin B at the time of the IOP increase was able to rescue the RGCs, as measured in a functional photopic electroretinogram, retinal nerve fiber layer thickness, and RGC counts. Mass-spectrometry-based proteomics and antibody-microarray measurements indicated that a α-crystallin injection distinctly up-regulated all of the subclasses (α, ß, and γ) of the crystallin protein family. The creation of an interactive protein network revealed clear correlations between individual proteins, which showed a regulatory shift resulting from the crystallin injection. The neuroprotective properties of α-crystallin B further demonstrate the potential importance of crystallin proteins in developing therapeutic options for glaucoma.

[Elevated Intraocular Pressure Induces Cellular Responses in the Retinal Capillaries]. / Erhöhter Intraokulardruck induziert primär zelluläre Reaktionen in den retinalen Kapillaren.

Background In the early diagnosis of clinical glaucoma, peripapillary bleedings were almost pathognomonic for a capillary insult. In the perfusion diagnostics, it is predominantly accepted that perfusion imbalances and IOP-induced changes occur and play a crucial role. Biomechanical peculiarities of the optic nerve head and cellular responses to astrocytes are also likely involved. Material and Methods We present in vivo and ex vivo models of IOP-elevation to enhance the resolution of examining cellular and molecular changes and to understand the mechanisms of capillary changes due to IOP-elevation. Results The in vivo model consists of cauterization-caused elevation of IOP in rat eyes. Two to 3 veins were cauterized to increase outflow resistance. The retinas were analyzed several weeks later and we found an abnormal expression of the neuron-specific molecule beta-III-tubulin in the capillary endothelium cells and in the vascular pericytes. Combined immunohistochemical stainings with different markers for various retinal cells confirmed the findings. The isolation of capillary endothelium cells and pericytes from rat brains (BMECs) and retinas (RMECs), and their cultivation under elevated IOP in vitro, confirmed the in vivo results. Conclusion The unexpected capillary response to elevation of IOP in vivo and in vitro could be seen as an early response of cells with expression of abnormal proteins. This result may explain clinical observations which dominate as peripapillary bleedings or microinfarctions and are likely associated with the glaucoma-induced opticopathy.

Does general anesthesia have a clinical impact on intraocular pressure in children?

BACKGROUND: Reliable measurement of intraocular pressure (IOP) is crucial in pediatric patients with suspected glaucoma. General anesthesia (GA) is usually needed in infants to allow a thorough examination. However, anesthesia itself may influence IOP, depending on the type used and the depth of sedation. The purpose of this study was to evaluate the normal distribution of IOP during GA in healthy children and to analyze differences in IOP relative to the anesthetics used and the measurement time point. METHODS: Approval for this observational study was received from the local institutional review boards and written informed consent was obtained from the children's parents. A total of 100 pediatric patients with no history of glaucoma scheduled for nonintraocular surgery underwent general anesthesia, induced with sevoflurane (s) or propofol (p) and maintained with either sevoflurane with remifentanil (S) or propofol with remifentanil (P). The patients were grouped to one of four subgroups (sS, sP, pP, pS) depending on the anesthetics used during induction and maintenance. Hemodynamic parameters and IOP were measured in both eyes at four defined time points: before anesthesia induction (M1); in apnea immediately after induction and before insertion of a laryngeal mask airway (M2); in deep anesthesia during mechanical ventilation (M3); and after extubation (M4), using a handheld Perkins applanation tonometer. Differences in IOP in both eyes during the measurement periods were analyzed using multivariate repeated-measures analysis of variance and Tukey-HSD as a posthoc test with statistical significance set at P < 0.05. Pearson correlation coefficient was used to investigate further relationships between heart rate, systolic blood pressure, and IOP. RESULTS: General anesthesia reduced IOP significantly. The mean IOP was normally distributed, with a mean of 7.4 ± 2.89 mmHg at M1. It decreased significantly to a minimum of 5.6 ± 3.04 mmHg (P < 0.01) at M2 and increased significantly to 7.2 ± 2.51 mmHg (P < 0.01) at M3 and again to 8.4 ± 3.72 mmHg (P = 0.03) at M4. All four subgroups (sS, sP, pP, pS) showed comparable decreases in IOP between M1 and M2. During deep anesthesia (M3) and during reversal (M4), the IOP increased again in all groups. During reversal (M4), however, the sS group had a significantly lower IOP than the pP group (P = 0.001) and sP group (P = 0.02). There were no correlations between changes in IOP and gender, age, or type of surgery. CONCLUSIONS: Sevoflurane and propofol, both in combination with remifentanil, significantly lower IOP in children. Individual IOP levels rise and fall during anesthesia, depending on the time point of measurement. The lowest IOP can be measured immediately after induction of anesthesia. This needs to be taken into account when measuring IOP in children.

Association between congenital nasolacrimal duct obstruction and delivery by cesarean section.

OBJECTIVE: Congenital nasolacrimal duct obstruction (CNDO) is the most common cause of neonatal epiphora. Persistence can lead to chronic dacryocystitis and amblyopia. This study analyzed the association between the incidence of CNDO and delivery by cesarean section. STUDY DESIGN: This was a retrospective cohort study of 386 children with CNDO (born between 2000 and 2008). The incidence of the delivery mode in patients with CNDO was compared with data from a corresponding population derived from annual birth statistics. RESULTS: There was no statistically significant association between the overall cesarean section rate and the incidence of CNDO, but primary cesarean section was significantly more frequent among patients with CNDO (73.15%, p < 0.05). The difference was significant for both genders for the period from 2000 to 2008 (p < 0.05%). The relative risk for CNDO was 1.7-fold increased in children delivered by primary cesarean section. CONCLUSION: Primary cesarean section may be a risk factor for CNDO.

Angiogenic gene therapy does not cause retinal pathology.

BACKGROUND: The potential negative influence of angiogenic gene therapy on the development or progression of retinal pathologies such as diabetic retinopathy (DR) or age-related macular degeneration (AMD) has led to the systematic exclusion of affected patients from trials. We investigated the role of nonviral fibroblast factor 1 (NV1FGF) in two phase II, multinational, double-blind, randomized, placebo-controlled, gene therapy trials (TALISMAN 201 and 211). METHODS: One hundred and fifty-two subjects with critical limb ischemia or claudication were randomized to receive eight intramuscular injections of 2.5 ml of NV1FGF at 0.2 mg/ml or 0.4 mg/dl or placebo. One hundred and fifty-two patients received a plasmid dose of NV1FGF of up to 32 mg or placebo. All patients underwent a systematic ophthalmologic examination at baseline and at 3, 6 or 12 months following gene therapy. Twenty-six of these patients (Münster subgroup) received a retinal fluorescence angiography at baseline and at final examination. RESULTS: Among those 26 patients, four of nine patients with diabetes suffered from nonproliferative DR. Three patients showed non-exsudative AMD. No change of retinal morphology or function was observed in Münster subgroup of both TALISMAN trials independent of the intramuscular NV1FGF dosage applied. CONCLUSIONS: Angiogenic gene therapy using NV1FGF is safe even in diabetics.

Correlation between early retinal nerve fiber layer loss and visual field loss determined by three different perimetric strategies: white-on-white, frequency-doubling, or flicker-defined form perimetry.

PURPOSE: To compare the significance of white-on-white standard automated perimetry (SAP), matrix frequency doubling technology (FDT), and flicker-defined form perimetry (FDF) for early detection of nerve fiber layer loss in early glaucoma patients. METHODS: Fifty-one healthy controls and 40 patients with early glaucomatous nerve fiber loss were enrolled in this study. Patients had retinal nerve fiber layer (RNFL) imaging and visual field testing by SAP, FDT matrix, and FDF perimetry at the same visit. Visual field defects were confirmed with two or more consecutive examinations by the same types of perimetry. Significant retinal nerve fiber layer loss and thus early glaucoma was defined with the reference to the RNFL thickness deviation map. The sensitivity, specificity, correlation, MD (mean deviation) and PSD (pattern standard deviation) visual field indexes, and area under the receiver operating characteristic curve (AUC) of MD and PSD of the perimetries were compared. RESULTS: There was a significant difference in nerve fiber layer thickness between healthy patients (97.7 ± 1.34 µm and patients with early glaucoma (84.1 ± 1.58 µm) (p < 0.001). Taking all patients with early glaucoma into consideration, the sensitivity was highest for FDF perimetry (87 %), followed by FDT matrix (62.5 %), and then SAP (40 %). The specificity was 69.2 % for SAP, 62.8 % for FDT matrix, and 38.4 % for FDF perimetry. MD (mean deviation) and PSD (pattern standard deviation) in FDF and FDT matrix were significantly different between patients with RNFL loss and those without (p < 0.05), while no difference could be found in SAP. The AUCs of MD followed a similar pattern, with FDF and FDT matrix perimetry having a suitable AUC of >0.6. AUCs of PSD were not reliable in all of the three VF devices. CONCLUSIONS: The sensitivity for detection of RNFL loss in early glaucoma seems to be higher in FDF and FDT matrix than SAP perimetry, while specifity was highest in SAP. Thus, simultaneous performance of FDF/FDT matrix and SAP perimetry seems beneficial for the correct diagnosis of early glaucoma in patients.

Cytokine Profiling in Aqueous Humor of Glaucoma Patients and in Retinas from an Ex Vivo Glaucoma Animal Model.

BACKGROUND: Although the current role of cytokines and neuroinflammation in glaucoma remains obscure, it represents an expanding field in research. The purpose of this study was to analyze cytokines in the aqueous humor (AH) of glaucoma patients and in retinas from an ex vivo glaucoma animal model, to aid in determining the role of neuroinflammation in glaucoma. METHODS: AH samples were collected from 20 patients during cataract surgeries (controls: n = 10, age = 70.3 ± 9.742; glaucoma: n = 10, age: 66.5 ± 8.073) in Shanghai East Hospital, an affiliate of Tongji University, between September 2018 and March 2019 and analyzed in duplicate by Luminex cytokine polystyrene color bead-based multiplex assay. Retinas from female Sprague-Dawley rats (n = 6) were harvested ex vivo and cultured with or without 60 mmHg of hydrostatic pressure for 24 hours. Retinal ganglion cells (RGCs) were quantified using Brn3a staining. Cytokines in the retina and culture medium were analyzed by rat cytokine array (Abcam). RESULTS: At baseline, patients with primary angle closure glaucoma (PACG) have significantly lower levels of IL-6 and IP-10 and a higher level of PDGF-BB in their AH, compared to the controls. Postoperatively, patients with PACG have significantly higher levels of IL-1ra, IL-13, and MIP-1α and a lower level of IL-6. Elevated hydrostatic pressure led to significant RGC loss in the retina, ex vivo, as well as the upregulation of ciliary neurotrophic factor (CNTF), IL-6, IL-10, IL-4, and TIMP-1 alongside the downregulation of PDGF-AA, MMP-8, TNF-α, and IFN-γ. Furthermore, eight cytokines were detected as being downregulated in the culture medium, including PDGF-AA, MMP-8, and IL-4. CONCLUSIONS: Proinflammatory cytokines showed changes in both AH and ex vivo. Further studies are needed on the role of these cytokines and their corresponding signaling pathways in both neurodegeneration and glaucoma.