scholarly journals A Porcine Ex Vivo Model of Pigmentary Glaucoma

2018 ◽  
Author(s):  
Yalong Dang ◽  
Susannah Waxman ◽  
Chao Wang ◽  
Ralitsa T. Loewen ◽  
Ming Sun ◽  
...  
Keyword(s):  
Trabecular Meshwork ◽  
Ex Vivo ◽  
Anterior Segment ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Ex Vivo Model ◽  
Trabecular Meshwork Cells ◽  
Gene Expression Microarrays ◽  
Expression Microarrays ◽  
And Behavior

Pigment dispersion can lead to pigmentary glaucoma, a poorly understood condition of younger myopic eyes with fluctuating high intraocular pressure. It has been difficult to investigate its pathogenesis without a model similar to human eyes in size and behavior. Here we present a porcine ex vivo model that recreates several features of pigmentary glaucoma, including intraocular hypertension, accumulation of pigment in the trabecular meshwork, and declining phagocytosis. We found that trabecular meshwork cells regulate outflow, form actin stress fibers, and have a decreased phagocytic activity. Gene expression microarrays and a pathway analysis of TM monolayers as well as ex vivo anterior segment perfusion cultures indicated that RhoA plays a central role in regulating the cytoskeleton, motility, and phagocytosis in the trabecular meshwork, providing new insights and targets to investigate in pigmentary glaucoma.

scholarly journals A porcine ex vivo model of pigmentary glaucoma

2017 ◽  
Author(s):  
Yalong Dang ◽  
Susannah Waxman ◽  
Chao Wang ◽  
Ralista T. Loewen ◽  
Ming Sun ◽  
...  
Keyword(s):  
Trabecular Meshwork ◽  
Ex Vivo ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Ex Vivo Model ◽  
Pigment Dispersion Syndrome ◽  
And Behavior ◽  
Human Eyes ◽  
Pigment Accumulation

AbstractPigment dispersion syndrome can lead to pigmentary glaucoma (PG), a poorly understood condition of younger, myopic eyes with fluctuating, high intraocular pressure (IOP). The absence of a model similar in size and behavior to human eyes has made it difficult to investigate its pathogenesis. Here, we present a porcine ex vivo model that recreates the features of PG including intraocular hypertension, pigment accumulation in the trabecular meshwork and relative failure of phagocytosis. Inin vitromonolayer cultures as well as inex vivoeye perfusion cultures, we found that the trabecular meshwork (TM) cells that regulate outflow, form actin stress fibers and have a decreased phagocytosis. Gene expression microarray and pathway analysis indicated key roles of RhoA in regulating the TM cytoskeleton, motility, and phagocytosis thereby providing new targets for PG therapy.

scholarly journals A Comparative Genome-wide Transcriptome Analysis of Glucocorticoid Responder and Non-Responder Primary Human Trabecular Meshwork Cells

2021 ◽  
Author(s):  
Kathirvel Kandasamy ◽  
Ravinarayanan Haribalaganesh ◽  
Ramasamy Krishnadas ◽  
Veerappan Muthukkaruppan ◽  
Colin E Willoughby ◽  
...  
Keyword(s):  
Ocular Hypertension ◽  
Trabecular Meshwork ◽  
Ex Vivo ◽  
Anterior Segment ◽  
Dexamethasone Treatment ◽  
Ex Vivo Model ◽  
Trabecular Meshwork Cells ◽  
Genome Wide ◽  
Contralateral Eye ◽  
Genome Wide Gene Expression

The genome-wide gene expression analysis of primary human trabecular meshwork (HTM) cells with known glucocorticoid (GC) responsiveness was not reported earlier. Therefore, the purpose of this study was to investigate genes and pathways involved in the GC responsiveness in human trabecular meshwork (HTM) cells using RNA sequencing. A perfusion cultured human anterior segment ex vivo model was utilized to identify the induction of GC-induced ocular hypertension in one eye of a paired eyes after dexamethasone treatment based on the maximum intraocular pressure response and in the contralateral eye, HTM cells were isolated to classify GC-responder and non-responder cells. Some previously reported and unique genes and their associated pathways were identified in HTM cells in response to dexamethasone treatment versus vehicle control and more significantly in GC-responder and non-responder cells. This study will open up the possibility of identifying suitable molecular targets which have the potential to treat GC-induced ocular hypertension/glaucoma.

scholarly journals Intraocular pressure elevation precedes a phagocytosis decline in a model of pigmentary glaucoma

2017 ◽  
Author(s):  
Yalong Dang ◽  
Susannah Waxman ◽  
Chao Wang ◽  
Ralitsa T. Loewen ◽  
Nils A. Loewen
Keyword(s):  
Intraocular Pressure ◽  
Trabecular Meshwork ◽  
Ex Vivo ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Ex Vivo Model ◽  
Phagocytic Capacity ◽  
Pressure Transducers ◽  
Intraocular Pressure Elevation ◽  
Iop Elevation

AbstractPurposeOutflow regulation and phagocytosis are key functions of the trabecular meshwork (TM), but it is not clear how the two are related in secondary open angle glaucomas characterized by an increased particle load. We hypothesized that diminished TM phagocytosis is not the primary cause of early ocular hypertension and recreated pigment dispersion in a porcine ex vivo model.Materials and MethodsSixteen porcine anterior chamber cultures received a continuous infusion of pigment granules (P), while 16 additional anterior chambers served as controls (C). Pressure transducers recorded the intraocular pressure (IOP). The phagocytic capacity of the trabecular meshwork was determined by fluorescent microspheres.ResultsThe baseline IOPs in P and C were similar (P=0.82). A significant IOP elevation occurred in P at 48, 120, and 180 hours (all P<0.01, compared to baseline). The pigment did not cause a reduction in TM phagocytosis at 48 hours, when the earliest IOP elevation occurred, but at 120 hours onward (P=0.001 compared to C). This reduction did not result in an additional IOP increase at 120 or 180 hours compared to the first IOP elevation at 48 hours (P>0.05).ConclusionIn this porcine model of pigmentary glaucoma, an IOP elevation occurs much earlier than when phagocytosis fails, suggesting that two separate mechanisms might be at work.

scholarly journals Impact of Pigment Dispersion on Trabecular Meshwork Cells

2018 ◽  
Author(s):  
Chao Wang ◽  
Yalong Dang ◽  
Ralitsa Loewen ◽  
Susannah Waxman ◽  
Priyal Shah ◽  
...  
Keyword(s):  
Trabecular Meshwork ◽  
Stress Fiber ◽  
Fiber Formation ◽  
Signaling Cascades ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Trabecular Meshwork Cells ◽  
Stress Fiber Formation ◽  
Ab Interno Trabeculectomy ◽  
And Migration

Purpose: To investigate the effect of pigment dispersion on trabecular meshwork (TM) cells. Methods: Porcine TM cells from ab interno trabeculectomy specimens were exposed to pigment dispersion, then analyzed for changes in morphology, immunostaining, and ultrastructure. Their abilities to phagocytose, migrate, and contract were quantified. An expression microarray, using 23,937 probes, and a pathway analysis were performed. Results: TM cells readily phagocytosed pigment granules. Pigment induced stress fiber formation (pigment (P): 60.1 &plusmn; 0.3%, n = 10, control (C): 38.4 &plusmn; 2.5%, n = 11, P &lt; 0.001) and contraction at 24 hours onward (P &lt; 0.01). Phagocytosis declined (P: 68.7 &plusmn; 1.3%, C: 37.0 &plusmn; 1.1%, n = 3, P &lt; 0.001) and migration was reduced after 6 hours (P: 28.0.1 &plusmn; 2.3, n = 12, C: 40.6 &plusmn; 3.3, n = 13, P &lt; 0.01). Microarray analysis revealed that Rho, IGF-1, and TGF&beta; signaling cascades were central to these responses. Conclusions: TM cell exposure to pigment dispersion resulted in reduced phagocytosis and migration, as well as increased stress fiber formation and cell contraction. The Rho signaling pathway played a central and early role, suggesting that its inhibitors could be used as a specific intervention in treatment of pigmentary glaucoma.

scholarly journals Intraocular pressure elevation precedes a phagocytosis decline in a model of pigmentary glaucoma

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 174 ◽  
Author(s):  
Yalong Dang ◽  
Susannah Waxman ◽  
Chao Wang ◽  
Priyal Shah ◽  
Ralitsa T. Loewen ◽  
...  
Keyword(s):  
Intraocular Pressure ◽  
Trabecular Meshwork ◽  
Ex Vivo ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Fluorescent Microspheres ◽  
Phagocytic Capacity ◽  
Pressure Transducers ◽  
Intraocular Pressure Elevation ◽  
Iop Elevation

Background: Outflow regulation and phagocytosis are key functions of the trabecular meshwork (TM), but it is not clear how the two are related in secondary open angle glaucomas characterized by an increased particle load. We hypothesized that diminished TM phagocytosis is not the primary cause of early ocular hypertension and recreated pigment dispersion in a porcine ex vivo model. Methods: Sixteen porcine anterior chamber cultures received a continuous infusion of pigment granules (Pg), while 16 additional anterior chambers served as controls (C). Pressure transducers recorded the intraocular pressure (IOP). The phagocytic capacity of the trabecular meshwork was determined by fluorescent microspheres. Results: The baseline IOPs in Pg and C were similar (P=0.82). A significant IOP elevation occurred in Pg at 48, 120, and 180 hours (all P<0.01, compared to baseline). The pigment did not cause a reduction in TM phagocytosis at 48 hours, when the earliest IOP elevation occurred, but at 120 hours onward (P=0.001 compared to C). This reduction did not result in an additional IOP increase at 120 or 180 hours compared to the first IOP elevation at 48 hours (P>0.05). Conclusions: In this porcine model of pigmentary glaucoma, an IOP elevation occurs much earlier than when phagocytosis fails, suggesting that two separate mechanisms might be at work.

scholarly journals Intraocular pressure elevation precedes a phagocytosis decline in a model of pigmentary glaucoma

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 174
Author(s):  
Yalong Dang ◽  
Susannah Waxman ◽  
Chao Wang ◽  
Priyal Shah ◽  
Ralitsa T. Loewen ◽  
...  
Keyword(s):  
Intraocular Pressure ◽  
Trabecular Meshwork ◽  
Ex Vivo ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Fluorescent Microspheres ◽  
Phagocytic Capacity ◽  
Pressure Transducers ◽  
Intraocular Pressure Elevation ◽  
Iop Elevation

Background: Outflow regulation and phagocytosis are key functions of the trabecular meshwork (TM), but it is not clear how the two are related in secondary open angle glaucomas characterized by an increased particle load. We hypothesized that diminished TM phagocytosis is not the primary cause of early ocular hypertension and recreated pigment dispersion in a porcine ex vivo model. Methods: Sixteen porcine anterior chamber cultures received a continuous infusion of pigment granules (Pg), while 16 additional anterior chambers served as controls (C). Pressure transducers recorded the intraocular pressure (IOP). The phagocytic capacity of the trabecular meshwork was determined by fluorescent microspheres. Results: The baseline IOPs in Pg and C were similar (P=0.82). A significant IOP elevation occurred in Pg at 48, 120, and 180 hours (all P<0.01, compared to baseline). The pigment did not cause a reduction in TM phagocytosis at 48 hours when the earliest IOP elevation occurred, but at 120 hours onward (P=0.001 compared to C). This reduction did not result in an additional IOP increase at 120 or 180 hours compared to the first IOP elevation at 48 hours (P>0.05). Conclusions: In this porcine model of pigmentary glaucoma, an IOP elevation occurs much earlier than when phagocytosis fails, suggesting that two separate mechanisms might be at work.

scholarly journals Replacement of the Trabecular Meshwork Cells—A Way Ahead in IOP Control?

Biomolecules ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1371
Author(s):  
Xiaochen Fan ◽  
Emine K. Bilir ◽  
Olivia A. Kingston ◽  
Rachel A. Oldershaw ◽  
Victoria R. Kearns ◽  
...  
Keyword(s):  
Stem Cells ◽  
Trabecular Meshwork ◽  
Vision Loss ◽  
Ex Vivo ◽  
Open Angle Glaucoma ◽  
Model Systems ◽  
Ex Vivo Model ◽  
Cell Replacement ◽  
Trabecular Meshwork Cells ◽  
Stem Cell Source

Glaucoma is one of the leading causes of vision loss worldwide, characterised with irreversible optic nerve damage and progressive vision loss. Primary open-angle glaucoma (POAG) is a subset of glaucoma, characterised by normal anterior chamber angle and raised intraocular pressure (IOP). Reducing IOP is the main modifiable factor in the treatment of POAG, and the trabecular meshwork (TM) is the primary site of aqueous humour outflow (AH) and the resistance to outflow. The structure and the composition of the TM are key to its function in regulating AH outflow. Dysfunction and loss of the TM cells found in the natural ageing process and more so in POAG can cause abnormal extracellular matrix (ECM) accumulation, increased TM stiffness, and increased IOP. Therefore, repair or regeneration of TM’s structure and function is considered as a potential treatment for POAG. Cell transplantation is an attractive option to repopulate the TM cells in POAG, but to develop a cell replacement approach, various challenges are still to be addressed. The choice of cell replacement covers autologous or allogenic approaches, which led to investigations into TM progenitor cells, induced pluripotent stem cells (iPSCs), and mesenchymal stem cells (MSCs) as potential stem cell source candidates. However, the potential plasticity and the lack of definitive cell markers for the progenitor and the TM cell population compound the biological challenge. Morphological and differential gene expression of TM cells located within different regions of the TM may give rise to different cell replacement or regenerative approaches. As such, this review describes the different approaches taken to date investigating different cell sources and their differing cell isolation and differentiation methodologies. In addition, we highlighted how these approaches were evaluated in different animal and ex vivo model systems and the potential of these methods in future POAG treatment.

scholarly journals Pigment dispersion syndrome with possible visual field loss (in Norwegian)

2010 ◽  
Vol 3 (1) ◽  
pp. 8-14
Author(s):  
Ellen Svarverud
Keyword(s):  
Trabecular Meshwork ◽  
Visual Fields ◽  
Anterior Segment ◽  
Clinical Signs ◽  
Visual Field Loss ◽  
Diagnostic Methods ◽  
Pigment Dispersion ◽  
Pigmentary Glaucoma ◽  
Pigment Dispersion Syndrome ◽  
Pigment Deposition

Pigment dispersion syndrome (PDS) is a condition of the anterior segment of the eye characterised by pigment deposition on a number of ocular structures. The condition is usually bilateral but most commonly asymmetric. In PDS, pigment is released from the posterior surface of the iris due to friction between the zonules and the iris. In itself, the condition does not represent any problems for the patient, but pigment deposition in the trabecular meshwork may interfere with drainage of the anterior chamber fluid and cause pigmentary glaucoma (PG). This case report presents a young myopic female with many of the classic signs of PDS; Krukenberg’s spindle, transillumination of mid-peripheral iris, deep anterior chambers, concave iris profile and increased pigmentation in the trabecular meshwork. Various diagnostic methods were applied to reveal clinical signs and to establish the patient’s visual function. Optic nerve heads and intraocular pressure were normal, but visual fields were suspect. The patient was advised to make an appointment with an ophthalmologist. Risk factors for development of PG and management of patients with PDS will be discussed.

scholarly journals Impact of freeze-thaw cytoablation on aqueous outflow patterns in ex vivo anterior chamber perfusion cultures and whole eyes

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 525
Author(s):  
Raoul Verma-Fuehring ◽  
Mohamad Dakroub ◽  
Alicja Strzalkowska ◽  
Piotr Strzalkowski ◽  
Hong Han ◽  
...  
Keyword(s):  
Trabecular Meshwork ◽  
Structural Changes ◽  
Ex Vivo ◽  
Anterior Segment ◽  
Large Change ◽  
Invasive Technique ◽  
Freeze Thaw ◽  
Before And After ◽  
Human Eyes ◽  
Porcine Eyes

Background: Porcine eyes have been widely used as ex vivo models in glaucoma research, as they share similar features with human eyes. Freeze-thawing is a non-invasive technique that has been used to obliterate living cells in anterior segment ex vivo cultures, to prepare them for further research such as cellular repopulation. This technique has previously been shown to reduce the intraocular pressure (IOP) in porcine eyes. The aim of this study was to investigate whether freeze-thaw cytoablation causes corresponding canalogram outflow changes in perfused anterior segment cultures (AFT) and whole porcine eyes (WFT). We hypothesized that the known IOP drop in AFT after trabecular meshwork ablation by freeze-thaw would be accompanied by a similarly large change in the distal outflow pattern. Methods: Two-dye (fluorescein and Texas red) reperfusion canalograms were used to compare the outflow time before and after two -80°C cycles of freeze-thaw. We assigned 28 freshly enucleated porcine eyes to four groups: perfused anterior segment dye controls (ACO, n = 6), perfused whole eye dye controls (WCO, n = 6), freeze-thaw treated anterior segment cultures (AFT, n = 10), and freeze-thaw treated whole eyes (WFT, n = 6). Results: In control groups ACO and WCO, the two different dyes had similar filling times. In AFT, the outflow pattern and filling times were unchanged. In WFT, the temporal superior quadrant filled more slowly (p = 0.042) while all others remained unchanged. The qualitative appearance of distal outflow spaces was altered only in some eyes. Conclusions: Freeze-thaw cytoablation caused neither loss nor leakage of distal outflow structures. Surprisingly, the loss of an intact trabecular meshwork over the entire circumference did not result in a general acceleration of quadrant outflow times. The results validate freeze-thawing as a method to generate an extracellular matrix without major structural changes.

scholarly journals Freeze-thaw decellularization of the trabecular meshwork in an ex vivo eye perfusion model

2017 ◽  
Author(s):  
Yalong Dang ◽  
Susannah Waxman ◽  
Chao Wang ◽  
Adrianna Jensen ◽  
Ralitsa T Loewen ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Trabecular Meshwork ◽  
Degrees Of Freedom ◽  
Ex Vivo ◽  
Fluorescent Microscopy ◽  
Anterior Segment ◽  
Perfusion Culture ◽  
Suicide Gene ◽  
Control Group ◽  
Freeze Thaw

Objective: Trabecular meshwork (TM) is the primary substrate of outflow resistance in glaucomatous eyes. Repopulating diseased TM with fresh, functional TM cells might represent a novel therapeutic breakthrough. Various decellularized TM scaffolds were developed by ablating existing cells with suicide gene therapy or saponin, but always with incomplete cell removal or dissolve the extracellular matrix. We hypothesized that a chemical-free, freeze-thaw method would be able to produce a fully decellularized TM scaffold for cell transplantation. Materials and Methods: We obtained 24 porcine eyes from a local abattoir, dissected and mounted them in an anterior segment perfusion and pressure transduction system within two hours of sacrifice. After they stabilized for 72 hours, eight eyes each were assigned to freeze-thaw (F) ablation (-80°C×2), to 0.02% saponin (S) treatment, or the control group (C), respectively. The trabecular meshwork was transduced with an eGFP expressing feline immunodeficiency viral (FIV) vector and tracked via fluorescent microscopy to confirm ablation. Following treatment, the eyes were perfused with standard tissue culture medium for 180 hours. We assessed histological changes by hematoxylin and eosin staining. TM cell viability was evaluated with a calcein AM/propidium iodide (PI) assay. We measured IOP and modeled it with a linear mixed effects model using a B-spline function of time with 5 degrees of freedom. Results: F and S experienced a similar IOP reduction by 30% from baseline (P=0.64). IOP reduction of about 30% occurred in F within 24 hours and in S within 48 hours. Live visualization of eGFP demonstrated that F conferred a complete ablation of all TM cells and only a partial ablation in S. Histological analysis confirmed that no TM cells survived in F while the extracellular matrix remained. The viability assay showed very low PI and no calcein staining in F in contrast to numerous PI-labeled dead TM cells and calcein-labeled viable TM cells in S. Conclusion: We developed a rapid TM ablation method that uses cyclic freezing that is free of biological or chemical agents and able to produce a decellularized TM scaffold with preserved TM excellular matrix in an organotypic perfusion culture.

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