Anterior Segment Of Human Eye Research Articles

Transplantation of iPSC-TM stimulates division of trabecular meshwork cells in human eyes

The trabecular meshwork’s (TM) physiological role is to maintain normal intraocular pressure by regulating aqueous humor outflow. With age, and particularly in eyes with primary open angle glaucoma, the number of cells residing within the TM is markedly decreased and the function of the tissue is compromised. Here we evaluate if transplantation of induced pluripotent stem cell derived TM like cells (iPSC-TM) restores TM cellularity and function in human eyes obtained from older human donors. Human iPSC were differentiated into iPSC-TM and compared to primary TM cells by RNAseq. iPSC-TM were then injected into the anterior segments of human eyes maintained in perfusion culture. Seven and 14 days eyes after injection eyes that received iPSC-TM contained significantly more cells in the TM. Fewer than 1% of all cells appeared to be iPSC-TM, but significantly more cells in these eyes were immunopositive for Ki 67 and incorporated BrdU. Our study demonstrates that transplantation iPSC-TM stimulates proliferation of endogenous TM cells in perfusion cultured human eyes from aged donors. These data, in concert with our previous findings in animal models, suggest that functional regeneration of the TM may be possible in human eyes with primary open angle glaucoma.

Intraocular Oxygen and Antioxidant Status: New Insights on the Effect of Vitrectomy and Glaucoma Pathogenesis

The purpose of this study was to investigate correlations of partial pressure of oxygen (pO2) in the ocular anterior segment of human eyes and aqueous humor antioxidant levels of ascorbate (AsA) and total reactive antioxidant potential (TRAP) with glaucoma and vitreous status. This prospective, cross-sectional study stratified patients (n= 288 eyes) by lens and vitreous status and the presence of primary open-angle glaucoma for statistical analyses. Intraocular pO2 concentrations were measured using a fiberoptic probe in patients at the beginning of planned glaucoma and/or cataract surgery. Aqueous humor specimens were obtained for antioxidant analysis of AsA and TRAP. Following prior pars plana vitrectomy, pO2 levels were significantly higher than in the reference group of cataract surgery in the anterior chamber angle (16.2 ± 5.0 vs. 13.0 ± 3.9mm Hg; P= .0171) and in the posterior chamber (7.6 ± 3.1 vs. 3.9 ± 2.7mm Hg; P < .0001). AsA and TRAP levels were significantly lower (1.1 ± 0.4 vs. 1.4 ± 0.5mM, respectively; 403.3 ±116.5 vs. 479.0 ± 146.7 Trolox units, respectively; P= .004 and P= .024, respectively) in patients after vitrectomy. In patients with an intact vitreous, neither pO2 nor antioxidant status correlated with lens status or glaucoma. Increased pO2 and antioxidant depletion following vitrectomy suggests an alteration of the intraocular oxidant-antioxidant balance. Our study links physiologic factors such as increased pO2 in the anterior chamber angle and the posterior chamber to decreased antioxidant levels in aqueous humor following vitrectomy. Oxidative stress/damage to the trabecular meshwork in such post-vitrectomy cases may contribute to intraocular pressure elevation and increased risk of glaucoma. NOTE: Publication of this article is sponsored bythe American Ophthalmological Society.

Polarization-sensitive optical coherence tomography imaging of the anterior mouse eye.

Polarization-sensitive optical coherence tomography (PS-OCT) enables noninvasive, high-resolution imaging of tissue polarization properties. In the anterior segments of human eyes, PS-OCT allows the visualization of birefringent and depolarizing structures. We present the use of PS-OCT for imaging the murine anterior eye. Using a spectral domain PS-OCT setup operating in the 840-nm regime, we performed in vivo volumetric imaging in anesthetized C57BL/6 mice. The polarization properties of murine anterior eye structures largely replicated those known from human PS-OCT imagery, suggesting that the mouse eye may also serve as a model system under polarization contrast. However, dissimilarities were found in the depolarizing structure of the iris which, as we confirmed in postmortem histological sections, were caused by anatomical differences between both species. In addition to the imaging of tissues in the anterior chamber and the iridocorneal angle, we demonstrate longitudinal PS-OCT imaging of the murine anterior segment during mydriasis as well as birefringence imaging of corneal pathology in an aged mouse.

Fluid and structure coupling analysis of the interaction between aqueous humor and iris.

BackgroundGlaucoma is the primary cause of irreversible blindness worldwide associated with high intraocular pressure (IOP). Elevated intraocular pressure will affect the normal aqueous humor outflow, resulting in deformation of iris. However, the deformation ability of iris is closely related to its material properties. Meanwhile, the passive deformation of the iris aggravates the pupillary block and angle closure. The nature of the interaction mechanism of iris deformation and aqueous humor fluid flow has not been fully understood and has been somewhat a controversial issue. The purpose here was to study the effect of IOP, localization, and temperature on the flow of the aqueous humor and the deformation of iris interacted by aqueous humor fluid flow.MethodsBased on mechanisms of aqueous physiology and fluid dynamics, 3D model of anterior chamber (AC) was constructed with the human anatomical parameters as a reference. A 3D idealized standard geometry of anterior segment of human eye was performed. Enlarge the size of the idealization geometry model 5 times to create a simulation device by using 3D printing technology. In this paper, particle image velocimetry technology is applied to measure the characteristic of fluid outflow in different inlet velocity based on the device. Numerically calculations were made by using ANSYS 14.0 Finite Element Analysis. Compare of the velocity distributions to confirm the validity of the model. The fluid structure interaction (FSI) analysis was carried out in the valid geometry model to study the aqueous flow and iris change.ResultsIn this paper, the validity of the model is verified through computation and comparison. The results indicated that changes of gravity direction of model significantly affected the fluid dynamics parameters and the temperature distribution in anterior chamber. Increased pressure and the vertical position increase the velocity of the aqueous humor fluid flow, with the value increased of 0.015 and 0.035 mm/s. The results act on the iris showed that, gravity direction from horizontal to vertical decrease the equivalent stress in the normal IOP model, while almost invariably in the high IOP model. With the increased of the iris elasticity modulus, the equivalent strain and the total deformation of iris is decreased. The maximal value of equivalent strain of iris in high IOP model is higher than that of in normal IOP model. The maximum deformation of iris is lower in the high IOP model than in the normal IOP model.ConclusionThe valid model of idealization geometry of human eye could be helpful to study the relationship between localization, iris deformation and IOP. So far the FSI analysis was carried out in that idealization geometry model of anterior segment to study aqueous flow and iris change.

Biometry of anterior segment of human eye on both horizontal and vertical meridians during accommodation imaged with extended scan depth optical coherence tomography.

PurposeTo determine the biometry of anterior segment dimensions of the human eye on both horizontal and vertical meridians with extended scan depth optical coherence tomography (OCT) during accommodation.MethodsTwenty pre-presbyopic volunteers, aged between 24 and 30, were recruited. The ocular anterior segment of each subject was imaged using an extended scan depth OCT under non- and 3.0 diopters (D) of accommodative demands on both horizontal and vertical meridians. All the images were analyzed to yield the following parameters: pupil diameter (PD), anterior chamber depth (ACD), anterior and posterior surface curvatures of the crystalline lens (ASC and PSC) and the lens thickness (LT). Two consecutive measurements were performed to assess the repeatability and reproducibility of this OCT. They were evaluated by calculating the within-subject standard deviation (SD), a paired t-test, intra-class correlation coefficients (ICC) and the coefficient of repeatability/reproducibility (CoR).ResultsThere were no significant differences between two consecutive measurements on either horizontal or vertical meridians under both two different accommodative statuses (P>0.05). The ICC for all parameters ranged from 0.775 to 0.998, except for the PSC (0.550) on the horizontal meridian under the non-accommodative status. In addition, the CoR for most of the parameters were excellent (0.004% to 4.89%). In all the parameters, only PD and PSC were found different between the horizontal and vertical meridians under both accommodative statuses (P<0.05). PD, ACD, ASC and PSC under accommodative status were significantly smaller than those under the non-accommodative status, except that the PSC at the vertical meridian did not change. In addition, LT was significantly increased when accommodation.ConclusionThe extended scan depth OCT successfully measured the dimensions of the anterior eye during accommodation with good repeatability and reproducibility on both horizontal and vertical meridians. The asymmetry of lens posterior surface and oval-shaped pupil were found during accommodation.

Anterior Segment Biometry during Accommodation Imaged with Ultralong Scan Depth Optical Coherence Tomography

To measure by ultralong scan depth optical coherence tomography (UL-OCT) dimensional changes in the anterior segment of human eyes during accommodation. Evaluation of diagnostic test or technology. Forty-one right eyes of healthy subjects with a mean age of 34 years (range, 22-41 years) and a mean refraction of -2.5 ± 2.6 diopters were imaged in 2 repeated measurements at minimal and maximal accommodations. A specially adapted and designed UL-OCT instrument was used to image from the front surface of the cornea to the back surface of the crystalline lens. Custom software corrected the optical distortion of the images and yielded the biometric measurements. The coefficient of repeatability and the intraclass correlation coefficient were calculated to evaluate the repeatability and reliability. Anterior segment parameters and associated repeatability and reliability upon accommodation. The dimensional results included central corneal thickness (CCT), anterior chamber depth and width (ACD, ACW), pupil diameter (PD), lens thickness (LT), anterior segment length (ASL = ACD + LT), lens central position (LCP = ACD + 1/2LT), and horizontal radii of the lens anterior and posterior surface curvatures (LAC, LPC). Repeated measurements of each variable within each accommodative state did not differ significantly (P>0.05). The coefficients of repeatability (CORs) and intraclass correlation coefficients for CCT, ACW, ACD, LT, LCP, and ASL were excellent (1.2%- 3.59% and 0.998-0.877, respectively). They were higher for PD (18.90%-21.63% and 0.880-0.874, respectively) and moderate for LAC and LPC (34.86%-42.72% and 0.669-0.251, respectively) in the 2 accommodative states. Compared with minimal accommodation, PD, ACD, LAC, LPC, and LCP decreased and LT and ASL increased significantly at maximal accommodation (P<0.05), whereas CCT and ACW did not change (P>0.05). The UL-OCT measured changes in anterior segment dimensions during accommodation with good repeatability and reliability. During accommodation, the back surface of the lens became steeper as the lens moved forward. The authors have no proprietary or commercial interest in any of the materials discussed in this article.

Ultra high-speed swept source OCT imaging of the anterior segment of human eye at 200 kHz with adjustable imaging range

We present an application of in vivo anterior segment imaging of the human eye with an ultrahigh speed swept source OCT instrument. For this purpose, a dedicated OCT system was designed and constructed. This instrument enables axial zooming by automatic reconfiguration of spectral sweep range; an enhanced imaging range mode enables imaging of the entire anterior segment while a high axial resolution mode provides detailed morphological information of the chamber angle and the cornea. The speed of 200,000 lines/s enables high sampling density in three-dimensional imaging of the entire cornea in 250 ms promising future applications of OCT for optical corneal topography, pachymetry and elevation maps. The results of a preliminary quantitative corneal analysis based on OCT data free form motion artifacts are presented. Additionally, a volumetric and real time reconstruction of dynamic processes, like pupillary reaction to light stimulus or blink-induced contact lens movements are demonstrated.

Meridional differences in internal dimensions of the anterior segment in human eyes evaluated with 2 imaging systems

To evaluate meridional differences in angle-to-angle (A-A) and sulcus-to-sulcus (S-S) dimensions in human eyes using very high frequency ultrasound (VHF US) and anterior segment optical coherence tomography (AS-OCT). John A. Moran Eye Center, University of Utah, USA; Berlin Eye Research Institute, Germany; Quattroelle Centri Oftalmochirurgici, Italy. Human autopsy eyes were fixated to special eye holders and had anterior segment scanning using 50 MHz ultrasound (Artemis, Ultralink) (20 eyes) and AS-OCT (SL-OCT, Heidelberg Engineering) (24 eyes). Seventy-three eyes (58 patients) also had anterior segment scanning using the same systems (Artemis, 50 eyes; SL-OCT, 23 eyes). Four meridians were evaluated in each eye. The A-A dimensions were measured by VHF US and AS-OCT. The S-S dimensions were also measured by VHF US. Statistical analyses were done with the Huynh-Feldt test. In the autopsy-eye study, the differences in A-A and S-S dimensions between the 4 meridians in eyes analyzed with VHF US were statistically significant (P=.002 and P=.009, respectively). The differences in A-A dimensions in eyes analyzed with the AS-OCT were also significant (P=.005). The same findings were observed in the clinical study (VHF US, P=.001 for A-A and P<.001 for S-S; AS-OCT, P<.001 for A-A). This study suggests that the internal geometry of the anterior segment of human eyes does not constitute a circle. This may have significant relevance for the accurate sizing of phakic intraocular lenses.

Evaluation of the Geometry of the Anterior Segment of Postmortem Human Eyes with two Imaging Systems

Purpose: High-frequency ultrasound and anterior segment optical coherence tomography (OCT) systems can be used to obtain non-invasive measurements of the anterior segment. The purpose of this study was to evaluate the geometry of the anterior segment of the human eye using these systems. Materials and Methods: Human eyes obtained postmortem were fixated to special eye holders and underwent anterior segment scanning using a 50MHz ultrasound system with digital enhancement (Artemis, Ultralink; 20 eyes), and an anterior segment OCT (SL-OCT, Heidelberg Engineering; 24 eyes). Four meridians were evaluated in each eye (vertical, horizontal, 135 and 45 degrees). Angle-to-angle dimensions were measured in the different meridians using the ultrasound and the OCT systems. Sulcus-to-sulcus dimensions were also measured in the different meridians using the ultrasound.

Perfusion of His-Tagged Eukaryotic Myocilin Increases Outflow Resistance in Human Anterior Segments in the Presence of Aqueous Humor

A previous study by the authors has shown that recombinant myocilin purified from a prokaryotic expression system increases outflow resistance in cultured human anterior segments. The present study was performed to determine whether full-length myocilin purified from a human trabecular meshwork cell expression system alters outflow resistance after infusion into human anterior segments. A feline immunodeficiency virus vector encoding both full-length myocilin (amino acids 1-503 fused to C-terminal V5 and six-histidine epitopes) and puromycin resistance was used to transduce a transformed trabecular meshwork cell line (TM5). Stably expressing cells were selected with puromycin. Recombinant myocilin was purified from the media using nickel ion affinity chromatography. Control purifications were performed on media from parental TM5 cells. Anterior segments of human eyes were placed in organ culture and perfused with either Dulbecco's modified Eagle's medium (DMEM) or DMEM supplemented with 50% porcine aqueous humor. One eye received an anterior chamber exchange with recombinant myocilin (2 microg/mL), whereas the fellow eye received an equal volume of control. Immunohistochemistry was performed with anti-myocilin and anti-V5 antibodies. Native polyacrylamide gel electrophoresis was used to analyze myocilin complex formation in porcine aqueous humor. Recombinant myocilin in porcine aqueous humor increased outflow resistance in cultured human anterior segments (91% +/- 68% [mean +/- SD] versus 18% +/- 31% in fellow control eye; n = 9, P = 0.004). Maximum outflow resistance was obtained 5 to 17 hours after infusion and remained above baseline for >3 days. Recombinant myocilin also increased outflow resistance in eyes incubated in DMEM, but only if myocilin was preincubated with porcine aqueous humor (78% +/- 77% when preincubated in DMEM containing porcine aqueous humor versus 13% +/- 15% when preincubated with DMEM alone, n = 6, P = 0.03). Recombinant myocilin appears to form a complex in porcine aqueous humor with a heat-labile protein(s). Immunohistochemistry revealed the presence of myocilin in the juxtacanalicular region of the trabecular meshwork. Myocilin purified from human trabecular meshwork cells increased outflow resistance in cultured human anterior segments, but only after incubation with porcine aqueous humor. Recombinant myocilin appears to form a complex in porcine aqueous humor that enables it to bind specifically within the trabecular meshwork.

Pharmacologic disruption of Schlemm's canal cells and outflow facility in anterior segments of human eyes.

To determine the effect of disruption of Schlemm's canal cells on outflow facility. Pharmacologic agents that weaken the cytoskeleton or interfere with integrin binding may allow targeted disruption of the cells lining Schlemm's canal because of the transmural pressure gradient the cells face as aqueous passes into the canal. Anterior segments of human eyes were placed in perfusion organ culture, and either single or sequential doses of H-7 or RGD peptide were added. Fellow eyes received vehicle or RGE peptide. Eyes were fixed and examined by light and electron microscopy. Both agents caused a partial loss of the endothelial lining of Schlemm's canal cells without disruption of trabecular cells in other regions. H-7 significantly increased outflow facility after single or sequential doses, with moderate cell loss of both the inner and outer wall canal cells: 20.0% +/- 10.5% of the width of the canal versus 5.2% +/- 3.7% in control meshworks (P = 0.05). No significant correlation between the amount of canal cell loss and outflow facility was found. RGD was associated with a variable loss of canal cells but did not change outflow facility. Pharmacologic disruption of Schlemm's canal cells appears possible. H-7 increased outflow facility, causing a partial loss of the endothelial lining of Schlemm's canal. A simple relationship between canal cells and outflow facility was not found; canal cells probably interact with the extracellular matrix in influencing outflow facility.

Effect of heparin II domain of fibronectin on aqueous outflow in cultured anterior segments of human eyes.

To determine whether an integrin/syndecan-binding domain of fibronectin, called the heparin II (Hep II) domain, affects outflow facility in the human eye. Anterior segments of human eyes were placed in perfusion organ culture. One eye of each pair received the Hep II domain, and the fellow eye received DMEM or a heat-denatured Hep II domain. The Hep II domain was produced as a recombinant glutathione S-transferase (GST)-fusion protein. Microscopic changes were assessed. Outflow facility in anterior segments treated with Hep II domain increased by 93% compared with that in anterior segments treated with DMEM. In contrast, facility in anterior segments treated with the heat-denatured Hep II domain showed very little change. Outflow facility remained high during Hep II domain perfusion and returned to baseline after removal of the protein. Electron microscopy revealed disruptions in the endothelial lining of Schlemm's canal in anterior segments fixed during maximum effect and in anterior segments after facility had returned to baseline. Scattered disruptions of canal cells were noted in control anterior segments. Trabecular cells in other regions looked normal. Major changes in the extracellular matrix of the juxtacanalicular tissue were not observed. Repeated doses of the Hep II domain administered after facility returned to baseline increased facility in two of three anterior segments. The Hep II domain of fibronectin increases outflow facility in the human anterior segment. This suggests that fibronectin-mediated interactions may have a role in modulating aqueous hydrodynamics. Such interactions may represent avenues of novel therapeutic interventions for glaucoma.

Distinctive distribution of HLA class II presenting and bone marrow derived cells in the anterior segment of human eyes.

Cells of bone marrow origin that normally occupy the stroma of the murine iris and ciliary body have been implicated in the immune phenomenon, anterior chamber associated immune deviation (ACAID). Following injection of antigen into the anterior chamber, cells of this type deliver an ACAID inducing signal into the systemic circulation, presumably through the outflow tract. In an effort to identify such cells in man, anterior chambers of 34 human donor eyes of different age groups were stained immunocytochemically with monoclonal antibodies directed at HLA class II molecules, CD 45 (a molecular marker of bone marrow-derived cells) and macrophage-associated membrane molecules (CD 68, CD 14). Within the outflow tissue, the cells of the filtering trabecular meshwork stained with none of those reagents. However, infrequent single, dispersed, dendritic cells were positively stained in the intertrabecular spaces. More numerous labelled cells were found in the anterior- and posterior-most portions of the non-filtering part of the trabecular meshwork. These cells were continuous with stained cells adjacent to the outer wall of Schlemm's canal and to the collector channels. Numerous labelled cells were seen in the vicinity of the intra- and episcleral vessels, the ciliary meshwork, the stroma of the ciliary muscle and epithelial processes, and the iris stroma. With advancing age, increasing numbers of CD 45+, HLA class II expressing cells appeared to accumulate in the so-called uveoscleral pathway. These results indicate that bone marrow-derived cells with the potential to function of ACAID induction reside within human eyes, and that cells of this type are located not only in the stroma of iris and ciliary body, but within the non-filtering portions of the trabecular meshwork and the uveoscleral pathway. The appearance of rare CD 45+ cells "in transit" in the filtering trabecular meshwork is compatible with the view that cells carrying ACAID-inducing signals to the systemic immune apparatus escape from the eye by this route.

Experimental errors in digital image processing for the determination of geometrical parameters of the human eye.

A new method is presented for measuring geometrical parameters of the anterior segment of human eye. A digital imaging technique is employed in conjunction with a slit lamp and TV camera. A method was developed to analyze images and the problem of minimizing measuring errors was studied. The method was tested using a contact lens to simulate the anterior corneal surface.