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.

Size does matter: what is the corneo-limbal diameter?

This review surveys available literature for corneal and limbal dimensions. With modern scleral lenses, these measures have become central to determining the overall size of a lens for satisfactory fitting. In general, published values are not based on a definition of what is being measured. In addition, the most widely accepted average corneal diameter measurement, 11.7 × 10.6 mm, emanates from one source published more than 50 years ago. This value was not based on a measurement but appears to be the cumulative impression of measurements from seven studies conducted between 67 and 127 years ago. Furthermore, in most instances, if at all stated, quoted measures are based on horizontal visible iris diameter, providing limited acknowledgement of limbal width and its inclusion as part of the corneal diameter. The corneo-scleral sulcus from one side to the other has been measured, giving a larger diameter, but may include at least part of the limbus. More objective measurements are possible with modern ophthalmic instrumentation but the lack of structural definition and low magnification resolution with these techniques raises concerns with the accuracy of the results. Measurement of the horizontal visible iris diameter does not include the limbal width, which means that the horizontal visible iris diameter is an underestimate of the true corneo-limbal diameter. This review concludes that the width of the limbus has been neither structurally defined nor accurately measured and that there is a need for the development of new protocols for determining the dimensions of the average cornea and limbus. It is predicted that more accurate measures will indicate that to vault across cornea with limbus and provide excellent comfort, the average cornea will need a lens to have a diameter of 16.0 mm or larger.

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.

Complications and fitting challenges associated with scleral contact lenses: A review.

INTRODUCTION: The modern scleral contact lens (ScCL) has evolved from the very first contact lens fitted 128 years ago. Originally manufactured in glass and oxygen impermeable plastics, these lenses are available today in high Dk gas permeable materials that allow permeation of oxygen, reducing many of the complications that were seen with older generation ScCL. However, as with any new contact lens modality, the modern ScCL brings with it a new set of complications and fitting limitations. METHODS: Pubmed searches under different keywords were conducted. RESULTS: Existing literature provides some reports of infection with the scleral devices, although these are often seen in severely compromised corneas, while hypoxic and inflammatory complications are rarely reported in the literature. Furthermore, the somewhat complex relationship of a scleral lens on the eye can create fitting and removal challenges. Anomalies such as conjunctival prolapse, epithelial bogging, midday fogging, and limbal bearing have been reported, and appear to be unique sequelae to scleral lens wear. CONCLUSION: Although this revolutionary technology broadens the scope in which practitioners can treat patients with irregular ocular surfaces, reports of these complications indicate that there is still a need for continued research to further enhance the clinical outcomes of this promising contact lens modality.

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.

Ultrastructure of UVR-B-induced cataract and repair visualized with electron microscopy.

PURPOSE: The aim of the study is to investigate and visualize the ultrastructure of cataract morphology and repair, after in vivo exposure to double threshold dose UVR-B in the C57BL/6 mouse lens. METHODS: Twenty-six-week-old C57BL/6 mice received in vivo double threshold dose (6.4 kJ/m2) UVR-B for 15 min. The radiation output of the UVR-source had λMAX at 302.6 nm. After a latency period of 1, 2, 4 and 8 days following UVR-B exposure, the induced cataract was visualized with electron microscopy techniques. Induced, cataract was quantified as forward lens light scattering. Damage to the lens epithelium and the anterior cortex was investigated with light microscopy in toluidine blue-stained semi-thin sections, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and dark field illumination photography. RESULTS: UVR-B-exposed lenses developed anterior subcapsular and/or cortical and nuclear cataract after 1 day. Lens light scattering peaked 2 days after exposure. Lens epithelial cell damage was seen in TEM as apoptotic cells, apoptotic bodies, nuclear chromatin condensation, and swollen and disrupted anterior cortex fibres throughout the sections of the whole anterior lens surface. These morphologic changes were also visualized with SEM. Within 8 days, anterior subcapsular cataract was repaired towards the anterior sutures. CONCLUSION: UVR-B exposure of double cataract threshold dose induces a subtotal loss of epithelial cells across the whole anterior surface of the lens. This damage to the epithelium is repaired by epithelial cell movement from the equator towards the lens sutures, thus in retrograde direction to regular epithelial cell differentiation.

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.

An explanation for the central to peripheral thickness variation in the mouse cornea.

BACKGROUND: The mouse corneal stroma varies in thickness across its diameter. The purpose of the present study was to explain this variation and to advance our understanding of stromal lamellar architecture in the mammalian cornea. METHODS: Eight C57BL/6 mice were killed, eyes enucleated, immersed in 2% glutaraldehyde fixative, processed and sectioned transversely for light and transmission electron microscopy. Transmission electron micrographs were assembled into montages and printed at 5000× magnification and used for lamellar counts and thickness assessments. RESULTS: The mouse cornea had an average of 49.8±2.4 lamellae centrally averaging 2.1µm in thickness versus 35.5±3.0 lamellae, averaging 1.9µm in thickness peripherally. The central to peripheral decrease in number lamellae and lamellar thickness measured utilizing the transmission electron microscope was statistically significant (P<0.005). CONCLUSIONS: This study demonstrated that the thickness difference between the thicker central and thinner peripheral mouse cornea is explained primarily by the number of lamellae present and that the peripheral lamellar dropout occurred in the anterior 2/3 of stroma. The decreased lamellar count towards the periphery suggested that not all lamellae cross the cornea limbus to limbus. These findings may be relevant to the thickness variation of the human cornea.