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OBJECTIVES: 1) To elucidate the role of collector channels in the aqueous humor outflow pathway 2) To suggest anatomic and functional methods of imaging collector channels in-vitro and in-vivo and 3) To discuss the role of such imaging modalities in the surgical management of glaucoma. METHODS: A thorough literature search was conducted on databases for studies published in English regarding the available methods to determine the role of collecting channels in normal and glaucomatous patients and to assess their patency. RESULTS: Intraocular pressure (IOP) exists as a balance between aqueous humor production and aqueous humor outflow. Collector channels are an essential anatomical constituent of the distal portion of the conventional aqueous humor outflow pathway. There are different surgical options for glaucoma management and with the recent advances in Schlemm's canal-based surgeries, collector channel's patency became a key factor in determining the optimum management for the glaucomatous eye. The advent of anatomic imaging methods has improved the ability to visualize collector channel morphology in-vitro, including swept-source optical coherence tomography (SS-OCT), spectral domain optical coherence tomography (SD-OCT), micro-computed tomography (micro CT), new immunohistochemistry techniques and scanning electron microscopy. The recent advent of real-time assessment of collector channel patency (including evaluation of episcleral venous outflow, observation of episcleral venous fluid wave, and tracer studies utilizing fluorescein, indocyanine green, and trypan blue) has been validated by the aforementioned anatomic imaging modalities. CONCLUSIONS: New modalities of in-vitro and in-vivo studies of collector channels provide promise to aid in the assessment of collector channel patency and individualization of surgical management for glaucoma patients.

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PURPOSE: To compare Schlemm canal (SC) and trabecular meshwork (TM) in children with healthy eyes and those with and without glaucoma after lensectomy. DESIGN: Cross-sectional observational study. PARTICIPANTS: Fifty children 4 to 16 years of age with healthy eyes and 48 children who underwent lensectomy (124 healthy and 72 postlensectomy eyes). METHODS: Anterior segment (AS) OCT (Tomey SS-1000 CASIA; Tomey, Nagoya, Japan) of the nasal iridocorneal angle at 2 levels of accommodative effort (2.5 diopters [D] and 15 D). For each parameter and state of accommodation, a random effects model was fitted to estimate differences between healthy eyes and eyes with history of lensectomy. MAIN OUTCOME MEASURES: Dimensions of SC and TM and conventional AS OCT iridocorneal angle measurements. RESULTS: The horizontal diameter of SC and its cross-sectional area (CSA) are significantly smaller in eyes that have undergone lensectomy versus healthy eyes. Accommodative effort increases SC size in healthy eyes, but not in eyes that have undergone lensectomy. CONCLUSIONS: Lensectomy is associated with a reduction in SC size and a loss of physiologic SC dilatation during accommodative effort, which may reflect a reduction in outflow facility and may contribute to the development of glaucoma after lensectomy.

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PURPOSE: To determine the effect of wearing a protective eye shield (mask) on limbal strain magnitude and variability in glaucoma eyes when sleeping with 1 side of the face down (FD) against a pillow. METHODS: A prospective, randomized, interventional trial was conducted at the Wilmer Eye Institute with 36 glaucoma patients. A contact lens sensor measured limbal strain (output in equivalent millivolts) during intervals of up to 60 minutes in lateral decubitus, FD, and supine positions. Eighteen subjects wore a mask during 1 of 2 FD intervals, with randomized assignment of the interval. Data from additional trials with no mask were included in some analyses. In addition, some facial-feature dimensions from 3D scanned images of 23 subjects were compared with limbal strain data. RESULTS: Wearing a mask trends toward a reduced mean change in contact lens sensor output (limbal strain) on moving to a FD positions [+34.1 mVeq, P=0.01 reduced by -22.3 mVeq, P=0.09 (n=36)]. Mask wearing reduced variability in strain while FD [-22.8 mVeq, P=0.04 (n=18)]. In eyes with past progressive visual field loss, the effect of the mask reduced mean strain change when moving to FD [-44.8 mVeq, P=0.02 (n=31)]. Longer corneal apex to nose-tip and to temple lengths were associated with reduced variability while FD [P=0.02 and 0.04, respectively (n=23)]. Treating both lengths as confounding factors increased statistical significance, particularly for analysis of the no-mask change in strain data moving to and from the FD position [P=0.004 to 0.002 and P=0.03 to 0.01 (n=23)]. CONCLUSION AND RELEVANCE: Wearing a mask reduced limbal strain and variation in limbal strain during simulated FD sleep, particularly in eyes with past field worsening, as did some facial features.

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BACKGROUND: Limbal stem cells (LSC) are progenitor cells in the ocular surface that renew the corneal epithelium. Limbal stem cell deficiency usually induces blindness through the loss of corneal transparency, and bilateral cases do not an accurate treatment because of the lack of an autologous source of stem cells. METHODS: Induced pluripotent stem cells (iPSC) are promising for use in cell therapy because of their autologous origin and the capability to differentiate into corneal epithelial cells. However, there are not standardized protocols to achieve a complete corneal epithelial differentiation. We examined the expression of several markers in a human episomal iPSC line after an induction period from embryoid bodies. RESULTS: Progenitor LSC and corneal epithelial differentiation markers, some extracellular matrix protein adhesion molecules, and wingless signaling pathway were studied. Overall, LSC progenitor and corneal epithelium differentiation markers increased after maintaining cell culture in specific conditions for 14 days, whereas pluripotency markers decreased. CONCLUSIONS: Our approach indicated that the optimal time point to initiate iPSC differentiation into LSC and corneal phenotypes, with the use of specific medium, is from 14 days after initial embryoid bodies treatment induction.

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PURPOSE: To study the effect of limbal relaxing incisions on corneal aberrations after phacoemulsification with aspheric intraocular lens (IOL) implantation. METHODS: Forty-five eyes underwent microincision cataract surgery with on-axis phacoemulsification incision. These eyes were divided into two groups: limbal relaxing incision group (23 eyes) and control group (22 eyes). In the limbal relaxing incision group, a single limbal relaxing incision was performed on the steepest meridian of corneal topography and the degrees of arc were determined using the DONO-Donnenfeld nomogram. The eyes in the control group were operated on by microincision cataract surgery without limbal relaxing incision. Corneal topography and wavefront analysis over 6-mm pupil size were performed preoperatively and 1 month postoperatively in both groups. Target induced astigmatism vector, surgically induced astigmatism vector, difference vector, magnitude of error, flattening effect, and torque were analyzed using the Alpins vectorial method. RESULTS: The limbal relaxing incision group showed significant reduction in topographic astigmatism by 51.87% of preoperative topographical astigmatism (P < .0001). The mean surgically induced astigmatism of limbal relaxing incision was 1.29 ± 0.71 diopters (D), which was close to the mean target induced astigmatism (1.33 ± 0.20 D), whereas the control group did not show a significant change in topographic astigmatism (0.17 ± 0.11 D; P = .73). Higher order aberrations of the cornea did not show significant change in either group except quatrefoil aberration, which significantly increased in the limbal relaxing incision group but not in the control group, with no significant difference between the two groups regarding corneal aberrations. CONCLUSIONS: Limbal relaxing incision is an effective method in reducing corneal astigmatism with neutral effect on corneal aberrations.

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PURPOSE: Previously, we demonstrated reduced Schlemm's canal cross-sectional area (SC-CSA) with increased perfusion pressure in a cadaveric flow model. The purpose of the present study was to determine the effect of acute IOP elevation on SC-CSA in living human eyes. METHODS: The temporal limbus of 27 eyes of 14 healthy subjects (10 male, 4 female, age 36 ± 13 years) was imaged by spectral-domain optical coherence tomography at baseline and with IOP elevation (ophthalmodynamometer set at 30-g force). Intraocular pressure was measured at baseline and with IOP elevation by Goldmann applanation tonometry. Vascular landmarks were used to identify corresponding locations in baseline and IOP elevation scan volumes. Schlemm's canal CSA at five locations within a 1-mm length of SC was measured in ImageJ as described previously. A linear mixed-effects model quantified the effect of IOP elevation on SC-CSA. RESULTS: The mean IOP increase was 189%, and the mean SC-CSA decrease was 32% (P < 0.001). The estimate (95% confidence interval) for SC-CSA response to IOP change was -66.6 (-80.6 to -52.7) µm(2)/mm Hg. CONCLUSIONS: Acute IOP elevation significantly reduces SC-CSA in healthy eyes. Acute dynamic response to IOP elevation may be a useful future characterization of ocular health in the management of glaucoma.

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The cornea is the clear tissue at the front of the eye that transmits light to the retina at the back of the eye. The cornea is covered by an epithelium and surrounded by a narrow band of tissue known as the limbus. The limbus has two important roles in maintaining a healthy corneal epithelium. First, stem cells for the corneal epithelium reside at the limbus and not in the cornea. Second, the limbus acts as a barrier separating the clear avascular corneal epithelium from the surrounding vascular conjunctival tissue. A failure of these limbal functions can result in the painful and blinding disease of limbal stem cell deficiency. In this disease, the corneal epithelium cannot be maintained by the stem cells, and the corneal surface becomes replaced by hazy conjunctival tissue. There are many causes of limbal stem cell deficiency, such as burns to the eye, inflammatory diseases, and hereditary diseases. Current understanding of the pathophysiology of the disease is discussed here. In particular, understanding whether the limbal stem cells are lost or become dysfunctional or indeed whether the limbal microenvironment is disturbed is important when developing appropriate management strategies for the disease.

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Limbal stem cell deficiency results from the loss of tissue regenerating stem and progenitor cells. Corneal epithelial regeneration is maintained by stem and progenitor cells which reside in the schlerocorneal limbus. They possess stem cell characteristics and can be stimulated to proliferate by external signals. The limbus is the stem cell niche for corneal epithelial stem cells and forms a unique microenvironment in which stem cell characteristics are conserved. Regulation of limbal epithelial stem cells is produced by a network of signals within the niche which governs cell fate decisions with regards to proliferation, differentiation or maintenance of a quiescent status.

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BACKGROUND AND OBJECTIVE: To determine the feasibility of attaching human amniotic membrane (HAM), pre-cultured with limbal stem cells (LSCs), to cornea using a novel, light-activated tissue bonding method. MATERIALS AND METHODS: LSCs were isolated from rabbit eyes, and then cultured on de-epithelialized HAM to create grafts (HAM/LSC). These were then transplanted onto rabbit eyes with surgically created limbal stem cell deficiency (LSCD). The grafts were secured either by sutures or by a light-activated method called photochemical tissue bonding (PTB). Outcomes included corneal opacity, inflammation, neovascularization, and collagen alignment. RESULTS: The isolated and cultured cells were verified to be LSCs based on their K19+/intergrin ß1+/P63+/K3 profile. Securing the HAM/LSC graft with PTB provided better outcomes. At 28 days post-surgery, the corneal opacity scores were significantly lower after securing the graft with PTB compared with suture attachment (0.8 ± 0.5 vs. 1.8 ± 0.5, P < 0.01). Similarly, neovascularization scores were lower after PTB (0.8 ± 0.5 vs. 1.5 ± 0.6, P < 0.01). Quantification of MPO and CD31 levels from immunofluorecent staining indicated that PTB stimulated less neutrophil infiltration (5.3 ± 2.2 vs. 13.3 ± 3.1, P < 0.01) and less new blood vessels formation (2.0 ± 0.8 vs. 6.3 ± 1.3, P < 0.01) at the wound site. The collagen alignment in PTB-treated corneas, as shown by immunofluorescence and second harmonic generation image, was better organized in the PTB-treated group than in the suture group. CONCLUSION: Bonding LSC grafts with PTB produced improved outcomes compared to suture attachment. This light-activated method is a promising modality for treating patients with LSCD.

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Caso clínico: Varón de 74 años de edad desarrolló una insuficiencia limbar parcial secundaria a una causticación en el ojo izquierdo. Fue tratado mediante la realización de una epiteliectomía sectorial conjuntival asociada a injerto de membrana amniótica. 3 semanas más tarde existía un epitelio corneal íntegro, estable y regular. Discusión: Este caso demuestra como la realización de una epiteliectomía sectorial asociada al injerto de membrana amniótica es un método eficaz y seguro para restaurar una superficie corneal estable en los casos de insuficiencia limbar parcial

Case report: A 74-year-old man developed a partial limbal stem cell deficiency secondary to a chemical injury in the left eye. He was treated with a sectorial conjunctival epitheliectomy and amniotic membrane transplantation. After 3 weeks there was an intact, smooth and stable corneal epithelium. Discussion: This case report emphasizes that conjunctival epitheliectomy and amniotic membrane transplantation appears to be a safe and effective method of restoring a stable corneal epithelium for cases of partial limbal stem cell deficiency

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BACKGROUND: To develop a new diagnostic technique to determine vertical ocular deviations when the center of the pupil is covered by swollen eyelids in up- and downgaze. PATIENTS AND METHODS: In upgaze (downgaze) the reflex of a diagnostic lamp held at about 50 cm distance from the patient is observed on the lower (upper) limbus. In the case of an asymmetric reflex, prisms are used to obtain symmetrical reflexes. The amount of prisms indicates the size of the vertical misalignment. In five healthy volunteers, the angles of vertical changes of gaze position were plotted against the prism size needed to recenter the perilimbal reflex. RESULTS: There was a linear correlation between the amount of upgaze changes in degrees and the strength of prisms used for compensation in degrees. This linear correlation was also found in downgaze. For both the correlation coefficient was r = 0.98 +/- 0.01. In upgaze the slope of the average regression line was 0.55 +/- 2.3 degrees, in downgaze - 4.1 +/- 0.8 degrees. A prism of 1 degrees corresponds in upgaze to a vertical deviation of about 1.3 +/- 0.14 degrees, in downgaze to a deviation of about 1.1 +/- 0.07 degrees. CONCLUSIONS: These results demonstrate that the perilimbal light reflex test is suitable for measuring simulated vertical ocular deviations. Therefore, the test may also be used in patients with vertical deviations who cannot be measured with classical methods. The method is more exact for measurements in upgaze.

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Frozen sections of corneoscleral buttons from normal and aniridic donors were stained with hematoxylin and periodic acid-Schiff, monoclonal antibodies AE-5 and AK-2 (to cornea-specific K3 and K12 keratins, respectively), and AM-3 (to conjunctival goblet cells) as well as with subtype-specific antibodies to seven different protein kinase C (PKC) subtypes, the signal transduction isoenzymes increasingly implicated in the regulation of cell growth and differentiation. Compared with the normal cornea, the aniridic cornea showed decreased AE-5 and AK-2 stainings, increased AM-3 staining, attenuated Bowman's membrane, invasion of new blood vessels, and limbal epithelial hyperplasia. In the normal tissue, the corneal epithelium expressed PKC alpha, lambda, and zeta; the limbal and conjunctival epithelia expressed additional PKC gamma. Conjunctival goblet cells expressed only PKC lambda. Within a given epithelium, different PKC subtypes had different cell-layer distributions. In the aniridic tissue, some of the four normally expressed subtypes were expressed in different cell layers, especially at the limbal region. PKC beta and PKC delta, which were normally weakly expressed, were markedly up-regulated. These results support the conclusion that the aniridic cornea does indeed manifest features of limbal stem cell deficiency with decreased corneal epithelial phenotype and increased conjunctival epithelial phenotype. Different capacities of proliferation and differentiation may be affected by the differential expression of PKC subtypes by different cell layers of normal ocular surface epithelia. The aberrant expression of PKC subtypes in aniridia may thus result in abnormal proliferation and differentiation noted in its ocular surface epithelia. Because limbal stem cells are the ultimate source of corneal proliferation and differentiation, we postulate that limbal deficiency in aniridia is a result of abnormal limbal stem cells.

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Infrared ocular thermograms were recorded for a group of 36 dry eye patients and for 27 age- and sex-matched controls. Mean ocular surface temperature was greater in the dry eye group (32.38 +/- 0.69 degrees C) compared with the control group (31.94 +/- 0.54 degrees C; p < 0.01). In addition, there was a greater variation of temperatures across the ocular surface in the dry eye group, illustrated by the difference in temperature between the limbus and the centre of the cornea (0.64 +/- 0.20 degrees C in dry eye patients compared with 0.41 +/- 0.20 degrees C in the control group; p < 0.001). This parameter was also shown to be greater in dry eye patients who displayed either a fast tear break-up time or a poor Schirmer's test result. Infrared thermography is a non-invasive and objective technique that may prove a useful research tool for study of the tear film, its deficiencies and its various treatment modalities.

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The cornea remains clear and avascular partly as a result of the compactness of the stromal lamellae. Loosening of these lamellae promotes loss of corneal transparency and peripheral vascularisation. Conversely, increased compactness of corneal tissue may possibly result in resorption of certain peripheral opacities. We report a case in which increased corneal compactness, produced by dellen formation, resulted in the disappearance of a type II Vogt's limbal girdle.

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