Central versus peripheral thickness in the human cornea explained.

PURPOSE: The human cornea is thicker in the periphery than the center and it has been suggested that this must be due to greater numbers of lamellae in the peripheral corneal stroma. The purpose of this study was to use high-resolution ultrastructural imaging to determine if the greater thickness of the peripheral cornea is due to the presence of more lamellae or if there is some other anatomical explanation. METHODS: In this study, full thickness corneas from three human donors were processed for light microscopy (LM) and transmission electron microscopy (TEM). Images were taken in three distinct stromal regions (anterior, middle, and posterior) from the central and peripheral cornea. Stromal thickness was evaluated by LM while TEM was used to evaluate numbers and thicknesses of lamellae, mean collagen fibril diameter, and mean collagen fibril density. RESULTS: Mean stromal thickness was significantly thinner in the central (415 ± 34 µm) compared to the peripheral (536 ± 29 µm) cornea (P = 0.009). Numbers of lamellae were not significantly different between central (246 ± 14) and peripheral (251 ± 14) cornea. Average lamellar thickness was not different across all regions of the cornea, except for the peripheral posterior where the lamellae were approximately 50 % thicker (P < 0.05). Collagen fibril diameters were larger in the peripheral cornea by approximately 30 % when compared to the central cornea, in all regions (P < 0.01). CONCLUSIONS: This study shows that it is an increase peripheral posterior lamellar thickness, rather than an increase in the number of lamellae, that accounts for the increase in corneal stromal thickness in the periphery of the human cornea. While collagen fibril diameters are greater throughout the peripheral stroma, the lamellae in the mid and anterior peripheral stroma are not thicker than centrally.

CLEAR - Anatomy and physiology of the anterior eye.

A key element of contact lens practice involves clinical evaluation of anterior eye health, including the cornea and limbus, conjunctiva and sclera, eyelids and eyelashes, lacrimal system and tear film. This report reviews the fundamental anatomy and physiology of these structures, including the vascular supply, venous drainage, lymphatic drainage, sensory innervation, physiology and function. This is the foundation for considering the potential interactions with, and effects of, contact lens wear on the anterior eye. This information is not consistently published as academic research and this report provides a synthesis from all available sources. With respect to terminology, the report aims to promote the consistent use of nomenclature in the field, and generally adopts anatomical terms recommended by the Federative Committee for Anatomical Terminology. Techniques for the examination of the ocular surface are also discussed.

A measure of tear inflow in habitual scleral lens wearers with and without midday fogging.

PURPOSE: The purpose of this pilot study was to evaluate tear inflow in a scleral lens system using fluorophotometry, and indirectly assess the exchange of the tear reservoir in habitual scleral lens wearers with the presence or absence of midday fogging (MDF). METHODS: Habitual scleral lens wearers (n=23) and normal scleral lens neophytes (n=10) were recruited. Of the 23 habitual wearers, 11 of them experienced MDF and 12 did not have a diagnosis of MDF. Contact lens-fitting characteristics were evaluated using ocular coherence tomograpy (OCT) and biomicroscopy. High molecular weight fluorescein (FITC) Dextran was instilled into the tear reservoir beneath the scleral lens, and the tear fluid fluorescein concentration was measured using the Fluorotron fluorophotometer. Calculated fluorescein concentrations were plotted over time to measure the fluorescein decay rate of the tear fluid beneath the scleral lens, which was used to calculate the tear exchange rate. RESULTS: There was no significant difference in tear inflow between the MDF group (mean: 0.111%) and the non-MDF group (mean: 0.417%), and there was a high amount of variability seen in the rates (p = 0.26). In addition, there was no significance between the tear reservoir thickness in the MDF (283um) and non-MDF (326um) groups (p = 0.53). CONCLUSIONS: The relationship between the amount of tear exchange during scleral lens wear and the incidence of MDF was not significant. Additional studies are needed to further examine the role of tear exchange in MDF and address the causes of variability to improve measurement techniques with fluorophotometry in the scleral lens system.

Assessing Scleral Contact Lens Satisfaction in a Keratoconus Population.

PURPOSE: To assess perceived comfort and related experiences of adapted keratoconic scleral contact lens (17-18.2 mm) wearers with a history of wearing other contact lens modalities and to compare these subjective clinical effects with previous experiences. METHODS: Twenty-four keratocones were recruited, signed an informed consent approved by the University Internal Review Board, and were asked to complete a survey and a dry eye questionnaire. RESULTS: Previous lens wearing experience included soft, piggy back, and hybrid lenses but predominantly was corneal gas permeable. An overwhelming majority strongly preferred the comfort and vision with the scleral contact lenses. This cohort of keratoconus patients were on average dry eye suspects according to a dry eye questionnaire but contained individuals with either normal or significantly dry eye scores. However, patients stated that they experienced less dryness with their scleral lenses, but slightly more than half of these patients experienced, at least occasionally, midday fogging. CONCLUSIONS: The scleral contact lenses are extremely well accepted by keratoconic patients because of comfort and vision these devices provide. For many patients, they offer further relief from dryness symptoms. However, midday fogging remains a limitation for many wearers.

Lamellar changes in the keratoconic cornea.

PURPOSE: The purpose of this study was to identify ultrastructural changes associated with ectasia and to determine the association between lamellar count and corneal thinning. METHODS: Five surgically removed keratoconic corneal buttons and four, non-keratoconic, normal eye bank control corneas were processed for transmission electron microscopy using an established protocol, ensuring minimal tissue distortion. A sequence of overlapping digital images, spanning the full apical cone corneal thickness, was assembled. A seamless digital montage was printed at 5000× magnification. Lamellae were counted in the anterior-posterior orientation, along a linear line, using established criteria for identification of individual lamellae. RESULTS: The stromal thickness estimated as a 95% confidence interval for the mean, CI (0.95), in the keratoconic corneas was 372 ± 62 µm, while in the normal cornea, it was 446 ± 89 µm. All keratoconic corneas showed ultrastructural evidence of lamellar splitting and a loss of interweaving anterior lamellae. In the keratoconic corneas, the median total linear stromal lamellar absolute count tangential to the corneal surface was 362, (25th percentile; 75th percentile) = (355; 365) lamellae and in the normal cornea, 246, (25th percentile; 75th percentile) = (239; 251). The linear lamellar density in the keratoconic corneas was estimated as CI (0.95) 117 ± 22 and 86 ±19 lamellae per 100 µm in the anterior and posterior portion of the stroma, respectively. In normal cornea, the linear lamellar density was estimated as CI (0.95) 51 ± 8 and 80 ± 20 lamellae per 100 µm. The mean difference of linear lamellar count between the anterior and the posterior portion of the cornea was estimated as CI (0.95) 31 ± 23 for keratoconic corneas and -29 ± 28 for the normal corneas. CONCLUSIONS: The current morphometric analysis of ultrastructural changes suggests that ectasia and thinning in keratoconus is associated with lamellar splitting into multiple bundles of collagen fibrils and loss of anterior lamellae. These structural changes, possibly in addition to lateral shifting of lamellae due to the pressure gradient over the cornea, are a potential explanation to the central loss of mass.

Contact lens care solution killing efficacy against Acanthamoeba castellanii by in vitro testing and live-imaging.

In the past decade there has been an increased incidence of Acanthamoeba keratitis, particularly in contact lens wearers. The aim of this study was to utilize in vitro killing assays and to establish a novel, time-lapse, live-cell imaging methodology to demonstrate the efficacy of contact lens care solutions in eradicating Acanthamoeba castellanii (A. castellanii) trophozoites and cysts. Standard qualitative and quantitative in vitro assays were performed along with novel time-lapse imaging coupled with fluorescent dye staining that signals cell death. Quantitative data obtained demonstrated that 3% non-ophthalmic hydrogen peroxide demonstrated the highest percent killing at 87.4% corresponding to a 4.4 log kill. The other contact lens care solutions which showed a 72.9 to 29.2% killing which was consistent with 4.3-2.8 log reduction in trophozoite viability. Both analytical approaches revealed that polyquaternium/PHMB-based was the least efficacious in terms of trophicidal activity. The cysticidal activity of the solutions was much less than activity against trophozoites and frequently was not detected. Live-imaging provided a novel visual endpoint for characterizing the trophocidal activity of the care solutions. All solutions caused rapid rounding or pseudocyst formation of the trophozoites, reduced motility and the appearance of different morphotypes. Polyquaternium/alexidine-based and peroxide-based lens care system induced the most visible damage indicated by significant accumulation of debris from ruptured cells. Polyquaternium/PHMB-based was the least effective showing rounding of the cells but minimal death. These observations are in keeping with care solution biocides having prominent activity at the plasma membrane of Acanthamoeba.

Recurrence or re-emergence of keratoconus--what is the evidence telling us? Literature review and two case reports.

Keratoconus may recur following penetrating or lamellar keratoplasty, but latency is considerably longer in the former. Since keratoplasty involves only partial excision of the cornea, and recent research strongly indicates the presence of the pathology in the peripheral host cornea, the reappearance of the pathology after a latency period is most likely due to migration of the disease from host to donor cornea. This notion is further corroborated by the shorter latency period in partial thickness keratoplasty, where more of the diseased host cornea remains in place. Other proposed causes for the recurrence of keratoconus, such as eye rubbing and contact lens wear, were reportedly not associated with a significant number of cases, and, therefore, are not the primary factor. Based on existing literature, it is concluded that, in post-keratoplasty keratoconus, the etiology stems from re-emergence of the disease rather than recurrence. Keratoconus patients in need of keratoplasty should be counseled on the possibility of the disease re-emerging.

Patient ocular conditions and clinical outcomes using a PROSE scleral device.

PURPOSE: To determine the type and distribution of ocular conditions cared for in a clinic dedicated to scleral devices and to report the clinical outcomes afforded by this approach. METHODS: Fifty-one charts of patients fitted unilaterally or bilaterally with a scleral device (Prosthetic Replacement of the Ocular Surface Ecosystem - PROSE) in a two year period were retrospectively reviewed. Patient demographics, ocular diagnoses, associated systemic conditions, best corrected visual acuity (BCVA) before and after fitting, Visual Function Questionnaire score (VFQ-25), and ocular surface disease index (OSDI) score were collected. RESULTS: All 51 patients were successfully wearing the PROSE device for a period of anywhere from weeks to years. The most common reasons for fitting were to relieve symptoms of moderate to severe dry eye syndrome ("DES", n=25), management of refractive problems ("refractive", n=23) with keratoconus being the most common (n=14), and to manage other anomalies ("other", n=3). Best corrected visual acuity (logMAR) improved with the wearing of the PROSE device for both the DES (17 letters) and the refractive group (10 letters), but not the "other" group. No serious complications were recorded for any of the patients. CONCLUSIONS: The PROSE device is a useful option not only for the management of ocular surface disease and optical imperfections, but also for other ophthalmic conditions. Moderate to severe dry eye was the most common anomaly managed, followed by eyes with irregular corneal astigmatism. DES and refractive patients experienced improvement in visual acuity with wearing of the PROSE device.

Central and peripheral corneal thinning in keratoconus.

PURPOSE: Recent data regarding the mid-peripheral portion of the removed corneal button in transplant surgery indicate histopathologically that keratoconus was present also in the peripheral portion of the button. The intent of this study was therefore to investigate if peripheral changes could also be detected clinically by measuring the central and peripheral corneal thickness of KC patients. METHODS: Corneal pachymetry was measured with the Visante optical coherence tomography and Orbscan II. Of 52 subjects, 26 were KC subjects and 26 were age- and sex-matched control subjects. RESULTS: Corneal thickness, in KC patients, was significantly reduced in all areas compared with corneal thickness among controls with both the Visante optical coherence tomography and Orbscan II. CONCLUSION: The results of the present study have provided clinical evidence that KC is a disease affecting a wider area of the cornea. Although the KC cornea shows the greatest change in thickness in the ectatic region, this attenuation declines toward the periphery. The presence of peripheral thinning indicated that KC may be a "pancorneal" pathology and not limited to the ectatic portion.

Morphologic alterations of the palpebral conjunctival epithelium in a dry eye model.

PURPOSE: To investigate the normal palpebral conjunctival histology in C57BL/6 mice and the structural changes that occur in a dry eye model. METHODS: Twenty-four male and female C57BL/6 mice, 8 untreated and 16 exposed to experimental ocular surface desiccating stress (DS). Ocular dryness was induced by administration of scopolamine hydrobromide (0.5 mg/0.2 mL) four times a day for 5 days (DS5) or 10 days (DS10). Counts and measurements were obtained using anatomical reference points, and goblet cell density was investigated with a variety of stains. RESULTS: Near the junction between the lid margin and the normal palpebral conjunctiva, the epithelium had an average thickness of 45.6 ± 10.5 µm, 8.8 ± 2.0 cell layers, versus 37.7 ± 5.6 µm, 7.4 ± 1.3 layers in DS10 (P < 0.05). In the goblet cell-populated palpebral region, the normal epithelium was thicker (P < 0.05) than on DS5 and DS10. In the control, 43% of the goblet cells were covered by squamous epithelium compared with 58% (DS5) and 63% (DS10) (P < 0.05). A decreased number of periodic acid-Schiff (PAS)-stained goblet cells and Alcian blue-stained goblet cells were observed in the dry eye. Not all goblet cells were stained with PAS and Alcian blue. CONCLUSION: The mouse palpebral conjunctival epithelium was structurally similar to the human. After DS, the palpebral conjunctival epithelium decreased in thickness and goblet cell access to the surface seemed to be inhibited by surrounding epithelial cells, potentially slowing down their migration to the surface. Differential staining with PAS and Alcian blue suggests that there may be different subtypes of conjunctival goblet cells.

Morphology of contact lens-induced conjunctival epithelial flaps: a pilot study.

PURPOSE: Conjunctival epithelial flap (CEF) is a conjunctival condition most commonly seen in silicone hydrogel contact lens wearers. This study utilized impression cytology to investigate the cellular composition and health of CEFs. METHODS: Nine (9) subjects were enrolled - 3 non-lens wearers, 3 contact lens wearers without CEF, and 3 contact lens wearers with CEFs wearing 8.4/13.8 lotrafilcon A lenses. Impression cytology samples were collected from the flap or similar locations, if CEF was absent, using rectangular 5 mm × 2 mm Millipore HAWP02500 membrane filters. The filters were gently pressed onto the conjunctiva, subsequently fixed in 95% alcohol, stained with hematoxylin-eosin and evaluated under an Olympus IX70 microscope. Measurements of the longest cell and nucleus dimensions were measured on 40 cells from each filter by utilizing NIH Image 1.63. RESULTS: CEF consisted of multilayers of epithelial and goblet cells and were devoid of inflammatory cells, basement membrane material and stromal tissue. The cytoplasmic and nuclear dimensions were similar within the groups and the cytoplasm-to-nucleus ratio was not different between the flap group and the non-lens wearing group. CONCLUSION: The CEF appeared to be formed by healthy epithelial and goblet cells that have been dislocated from their normal location along the conjunctival surface by the lens edge. No inflammatory cells were present in this contact lens induced condition, which is reported to be associated most commonly with the silicone hydrogel material.

Does the eye benefit from wearing ultraviolet-blocking contact lenses?

OBJECTIVES: The human eye is exposed to toxic ultraviolet radiation (UVR) from sunlight and artificial sources. The UVR-induced damage occurs in ocular tissues from the corneal surface to the retina. Although the cornea and crystalline lens provide inherent UVR protection, the anterior ocular surface and the limbus, which contains stem cells, receive toxic levels of UVR from relatively short solar exposures. METHODS: Shading headwear and some UVR-blocking sunglasses are designed to reduce direct solar exposure but may not protect the eye from diffuse ambient and surface reflected light. If the squint mechanism is reduced because of the reduction of visible light, the ocular surface is then exposed to ambient and reflected UVR. In addition, laterally incident radiation that is focused across the cornea onto the limbus, a phenomenon known as peripheral light focusing, can increase the dose at the nasal limbus by a factor of 20. RESULTS: The UVR-blocking contact lenses that cover the limbus provide protection from all sources of ocularly incident UVR. Although directly relating solar UVR dose to ocular damage is epidemiologically challenging, irradiation of ocular cell cultures can estimate the toxic effects of UVR exposure. The use of UVR-blocking contact lenses greatly increases the time the wearer can be exposed to solar UVR before a toxic ocular dose is reached. CONCLUSIONS: There is a need for the development of a scientifically rigorous, clinically applicable ocular protection factor metric, based not only on the transmittance of eyewear but on the protection afforded from the total UVR field and the length of that exposure.

Quantified histopathology of the keratoconic cornea.

PURPOSE: This study systematically investigated and quantified histopathological changes in a series of keratoconic (Kc) corneas using a physiologically formulated fixative to not further distort the already distorted diseased corneas. METHODS: Twelve surgically removed Kc corneal buttons were immediately preserved and processed for light and transmission electron microscopy using an established corneal protocol. Measurements were taken from the central cone and peripheral regions of the host button. The sample size examined ranged in length from 390 to 2608 µm centrally and 439 to 2242 µm peripherally. RESULTS: The average corneal thickness was 437 µm centrally and 559 µm peripherally. Epithelial thickness varied centrally from 14 to 92 µm and peripherally from 30 to 91 µm. A marked thickening of the epithelial basement membrane was noted in 58% of corneas. Centrally, anterior limiting lamina (ALL) was thinned or lost over 60% of the area examined, whereas peripheral cornea was also affected but to a lesser extent. Histopathologically, posterior cornea remained undisturbed by the disease. Anteriorly in the stroma, an increased number of cells and tissue debris were encountered, and some of these cells were clearly not keratocytes. CONCLUSIONS: It is concluded that Kc pathology, at least initially, has a distinct anterior focus involving the epithelium, ALL, and anterior stroma. The epithelium had lost its cellular uniformity and was compromised by the loss or damage to the ALL. The activity of the hitherto unreported recruited stromal cells may be to break down and remove ALL and anterior stromal lamellae, leading to the overall thinning that accompanies this disease.

In vitro effects of medium tonicity, nutrient concentration, and free chlorine content on Acanthamoeba.

BACKGROUND: The environment preferred by Acanthamoeba trophozoites and the mechanism by which the amebae enters the cornea are not yet fully understood. A better understanding of the pathogenesis of this disease may help with prevention and treatment. PURPOSE: To define the preferred environments for Acanthamoeba survival and proliferation in vitro by examining the effect of tonicity, nutrient concentration, and free chlorine content on Acanthamoeba. MATERIALS AND METHODS: Human corneal isolates of Acanthamoeba castellanii and Acanthamoeba polyphaga trophozoites were cultured at 22°C (room temperature) in PYG (peptone-yeast extract-glucose) medium. The effect of tonicity on amebae was determined by incubating trophozoites in sodium chloride solutions in concentrations ranging from 0% to 10% for 19 days. Two different sets of media were prepared-one with and the other without added nutrients. The tonicity varied from 50 to 3438 mOsm/L while the pH was maintained at 6.7-6.8. Aliquots were recovered to determine the number and morphologic type of the amebae. To test the effect of chlorine, Acanthamoeba trophozoites were incubated for 7 days in buffered solutions with free chlorine concentrations varying from 0 to 5 mg/L free chlorine at 22°C. The pH was maintained at 7.2 and the tonicity varied from 88 to 92 mOsm/L. Trophozoites were enumerated by hemocytometer. RESULTS: Low tonicity solutions (<300 mOsm/L) favored the trophozoite stage, but elevating tonicity encouraged encystment. Only 3.3-3.9% of the trophozoites remained in 10% NaCl, while 46-58% of the trophozoites were present in distilled water. Increasing osmolality yielded a smaller number of Acanthamoeba with a greater proportion of cysts. Nutrients improved the replication rate at lower concentrations, increased the number of trophozoites and reduced the percentage of cysts. Chlorine completely inhibited both species of Acanthamoeba at free chlorine levels of 5mg/L, while lesser concentrations were less inhibitory. CONCLUSIONS: Acanthamoeba prefer hypotonic environments. Nutrients merely slowed the conversion of trophozoites to cysts at higher tonicity levels. Chlorine concentrations less than 5 mg/L, ocular irritation level, did not effectively convert trophozoites into cysts. We conclude that contact lens patients should avoid hypotonic ocular exposures, especially tap water and stagnant media such as lake water, and water from poorly maintained swimming pools and hot-tubs.

Current silicone hydrogel UVR blocking lenses and their associated protection factors.

PURPOSE: Ultraviolet radiation (UVR) blocking contact lenses provides ocular protection factors (PF) that vary with lens thickness and the nature of the polymer dopant. This study measured the UVR PF of silicone hydrogel lenses outdoors and compares this to known methods for determining PF mathematically. METHODS: Ambient measurements were taken using an UVA-B radiometer, adapted to hold the test lenses over its sensor to quantify their UVR blocking capabilities. The UVR blocking silicone hydrogel lenses tested included galyfilcon A, senofilcon A, and enfilcon A. The first UVR blocking hydrogel lens, vasurfilcon A, and a non-blocking silicone hydrogel lens, lotrafilcon B, served as a comparison. Lens transmittance at the centre and periphery was measured and PF calculated to predict signal reduction for comparison with field measurements. RESULTS: There was a significant range of cut-off wavelengths across the lenses, ranging from 370nm for enfilcon A to 390nm for vasurfilcon A, with lotrafilcon B transmitting down to 265nm, with a 3nm shift from centre to periphery across the -3.00 D UVR blockers. The UVR reduction calculated from the transmittance data correlates well with field data, ranging from 90-98% for the UVR blockers to 13% for the non-blocker. CONCLUSIONS: The silicone hydrogel lenses showed a wide range of transmittance curves with increasing PF from centre to periphery. PF calculations work well but do not always compare precisely with measured data due to factors such as sensor spectral response and the nature of the incident solar spectrum.