Next-Generation Sequencing of the Ocular Surface Microbiome: In Health, Contact Lens Wear, Diabetes, Trachoma, and Dry Eye.

OBJECTIVES: To assess publications examining the occurrence, composition, and clinical significance of a microbiome at the ocular surface. METHODS: MEDLINE, EMBASE, and Google Scholar were searched. Reference lists of included articles were also searched for relevant citations. All publications up to June 1, 2019, were analyzed. RESULTS: Eleven articles and 1 abstract were included, analyzing 661 patients. Articles generally report bacteria to the genus level. The presence of DNA associated with diverse bacterial species was reported including pathogenic species, such as Pseudomonas and Neisseria. Bacterial DNA that makes up the microbiome, such as Acinetobacter, Actinomyces, Aquabacterium, Bradyrhizobium, Corynebacterium, Sphingomonas, Staphylococcus, and Streptococcus, in other parts of the body was found. The putative ocular microbiome is consistent between right and left eyes and is affected by contact lens use (higher Pseudomonas levels) and blepharitis (higher Staphylococcus levels). CONCLUSIONS: There is a significant likelihood that there is at least a transitory ocular surface microbiome, with Acinetobacter, Corynebacterium, Propionibacterium, Staphylococcus, and Streptococcus detected in at least 7 of 11 studies. However, further investigation attempting to control for environmental and methodological contaminants (Aquabacterium and Bradyrhizobium are commonly identified as contaminants in DNA extraction kits) is required. Bacteria, such as Propionibacterium, Staphylococcus, and Streptococcus, capable of causing sight-threatening infections may reside on a healthy ocular surface. With greater understanding, we can establish whether elements of the ocular surface microbiome are harmful or protective (despite their small quantities); furthermore, new therapeutic agents can be identified to treat and prevent ocular surface infection and inflammation.

Impact of First Healthcare Provider on Acanthamoeba Keratitis Course: How to Overcome Poor Prognosis in Acanthamoeba Keratitis Treatment? A Single Tertiary Center, Observational Study.

Background: To assess the difference in course and final visual outcome of Acanthamoeba keratitis (AK) patients based on the first healthcare provider (HCP) seen. Methods: Retrospective observational cohort study of AK patients admitted to the Manchester Royal Eye Hospital between 2003 and 2017. HCPs were grouped (Group 1: Optometrists, Opticians; Group 2: General Practitioners (GPs); Group 3: Ophthalmologists) and the data analyzed on demographics, risk factors, clinical history, clinical features, and Acanthamoeba subspecies. Results: Forty-one patients with unilateral culture-proven AK were included. Median time to consultation with first HCP was 7 days (IQR 4-14 days), while mean time to the correct diagnosis of AK was 15 days (IQR 7-29 days). Patients saw an optician, optometrist or ophthalmologists significantly earlier than GPs (median 4 days, vs 15 or 5 days, respectively, p = 0.04). Bacterial keratitis was the most common initial clinical diagnosis (43%). The shortest time to making the AK diagnosis (median 11 days) and the highest rate of initiating AK treatment started at the first visit (38%) were both in the ophthalmologists' group. No significant differences were observed in initial and final visual acuity between HCP groups (p = 0.36). Conclusion: AK patients often seek ocular help earlier from optometrists and opticians than medical doctors. Final clinical outcomes did not significantly differ based on the first HCP seen, but ophthalmologists were more likely to make the diagnosis of AK and initiate anti-amoebal therapy faster than other HCPs. Greater education and collaboration between ophthalmologists and other HCPs to increase awareness of AK are needed.

Corneal epithelialisation on surface-modified hydrogel implants: artificial cornea.

The objective was to investigate corneal re-epithelialisation of surface-modified polymethacrylate hydrogel implants in order to evaluate them as potential materials for an artificial cornea. Polymethacrylate hydrogels were modified with amines and then coated with different extracellular matrix proteins (collagen I, IV, laminin and fibronectin). The modified hydrogels were surgically implanted into bovine corneas maintained in a 3-D culture system for 5 days. The epithelial growth across the implant surface was evaluated using fluorescent, light and electron microscopy. Full epithelialisation was achieved on 1,4-diaminobutane-modified hydrogels after coating with collagen IV. Hydrogels modified with 1,4-diaminobutane but without further coating only showed partial re-epithelialisation. Hydrogels modified with other amines (1,2-diaminoethane or 1,3-diaminopropane) showed only partial re-epithelialisation; further coating with extracellular matrix proteins improved epithelialisation of these surfaces but did not result in complete re-epithelialisation. Evaluation of the corneas implanted with the 1,4-diaminobutane-modified hydrogels coated with collagen IV showed that the artificial corneas remain clear, integrate well and become covered by a healthy stratified epithelium. In conclusion the 1,4-diaminobutane surface-modified hydrogel coated with collagen IV supported the growth of a stable stratified epithelium. With further refinement this hydrogel has the potential to be used clinically for an artificial cornea.

Microbial Keratitis in Corneal Transplants: A 12-Year Analysis.

PURPOSE: To investigate the frequencies, trends, and in vitro drug susceptibilities of the causative pathogens in corneal transplant microbial infections in Manchester Royal Eye Hospital. METHODS: Corneal scrape results recorded by the microbiology service between 2004 and 2015 were extracted from an established database. This microbiological data was matched with a separate database of all corneal transplant procedures performed in our centre over this time period. Patient records were examined to collect specific patient data and to confirm the diagnosis of microbial keratitis. RESULTS: A total of 1508 grafts had been performed at our centre in this period. 72 episodes of graft microbial keratitis were identified from 66 eyes that had undergone keratoplasty procedures. Mean age was 56, and 51% of subjects were male. Ninety-three percent of microbial keratitis episodes occurred in penetrating keratoplasty procedures and 6% in deep anterior lamellar keratoplasty procedures. No endothelial grafts presented with infections throughout this time period. Of the 79 organisms identified, 73% were gram positive, 23% gram negative and 4% fungi. With regard to gram-positive organisms, vancomycin and gentamicin showed 100% and 91% susceptibility, respectively. Ofloxacin had a resistance rate of 13.7%. In terms of gram-negative organisms, gentamicin and chloramphenicol showed 100% sensitivity, with cefuroxime showing 69%. Resistance rates were less than 15% in all tested gram-negative antimicrobials. CONCLUSION: This paper describes the largest collection of corneal transplant infections identified within the UK. This finding may aid clinicians in predicting possible causative organisms for microbial keratitis and aid antibiotic choice.

The importance of grating orientation in contrast sensitivity following refractive surgery.

Higher-order aberrations degrade visual performance when the pupil is large, particularly in eyes that have abnormally high amounts of aberrations, such as those that have undergone refractive surgery. Geometrical optics predicts that asymmetrical aberrations such as coma can produce orientation selective effects, much like uncorrected astigmatism. Coma is also one of the main aberrations to increase following refractive surgery. Orientation may therefore be an important parameter when testing grating based contrast sensitivity, particularly in refractive surgery patients. Contrast sensitivity to four orientations of a 12 cycles deg(-1) sine wave grating was measured in normals (n = 34) and refractive surgery patients (n = 12). In over a third of normal subjects the higher-order aberrations produced a significant orientation-specific change in contrast sensitivity (13 out of 34 eyes, p < 0.05). No significant differences existed in the aberrations between those that displayed orientation-selective changes and those that did not. In subjects who underwent refractive surgery those that displayed orientation selective changes had significantly higher amounts of overall aberrations and also higher amounts of primary coma than those that did not. These results indicate that grating orientation is an important factor when assessing the effects of higher-order aberrations on contrast sensitivity.

Contrast sensitivity based orientation changes in refractive surgery patients.

It is well known that refractive surgery leads to an increase in higher-order aberrations. These can cause problems such as glare, starburst and halos in vision, particularly in low light levels when the pupil is large. The overall wavefront aberration of the eye is typically non-circularly symmetric due to the presence of aberrations such as coma and trefoil. These asymmetries have been shown to induce orientation-selective effects in grating-based contrast sensitivity tests. Classically, contrast sensitivity is assumed to be independent of orientation and is measured at only one orientation. Examining more orientations may give a better indicator of visual performance when abnormal amounts of aberration are present. To examine this, contrast sensitivity through a dilated pupil was determined for four orientations of a 12 cycles deg(-1) sine wave grating in 12 subjects both before and after refractive surgery. Surgery produced orientation-selective changes in contrast sensitivity in most patients (9 out of 12, p < 0.05). The correlation between contrast sensitivity and higher-order aberrations was found to improve when several orientations were tested compared with data from only one orientation (combined orientations, r(2) = 0.46, p = 0.007; single orientation r(2) = 0.55, p < 0 .05 to r(2) = 0.09, p > 0.05). These results indicate that grating orientation is an important factor when investigating the link between visual performance and higher order aberrations.

Corneal Imaging and Densitometry Measurements in Juvenile and Adult Keratoconus Patients to Evaluate Disease Progression and Treatment Effects After Corneal Cross-Linking.

PURPOSE: In this study densitometry software for the Oculus Pentacam was used to investigate the treatment outcomes of corneal cross linking (CXL) in adult and juvenile keratoconus (KCN) patients. Densitometry measurements were taken before and after treatment and followed up for one year. METHODS: A comparative study was carried out at Manchester Royal Eye Hospital. Corneal densitometry measurements collected before and after CXL treatment for 32 eyes from KC patients, aged between 12 and 39, were divided to 2 groups 13-18 years (juvenile group) and 19-39 years (adult group) and analysed and compared to pre and post treatment at 3, 6 and 12 months for each group and between both groups. RESULTS: Analysis of densitometry measurements found higher corneal densitometry after CXL which peaks at three months post treatment in both groups. There was significant diversity in corneal densitometry measurements in the stromal zone 0-2 and 2-6 mm for all layers except the posterior layer for both groups (P<0.05). Significantly increased densitometry value was found higher in the juvenile group at six months in the central (P=0.006) and posterior (P=0.004) layers for zone 0-2 mm. The same layers differed significantly also in the 2-6 mm zone in all layers (P=0.01). One year post treatment the same significant increased densitometry level was seen in the juvenile group in the 0-2 mm zone of the central (P=0.007) and posterior layers (P=0.01), as was the 2-6 mm zone (P=0.04). However, no significant difference was found between pre and post treatment for best corrected visual acuity (BCVA), central corneal thickness (CCT) and thinnest area between both groups. A significant difference was found between pre and post treatment for best corrected visual acuity (BCVA), in the adult group at 6 and 12 months post-treatment from pre-treatment (P=0.02, P=0.16) respectively. CONCLUSION: Corneal clarity post CXL treatment in the juvenile group differed significantly from the adult group. Both groups showed increased haze at 3 months post treatment but the adults showed improvement over the next 9 months. In contrast, the juvenile group showed higher densitometry readings at both 6 and 12 months post treatment in comparison to adult group. The reasons for this remain unclear.

Importance of grating orientation when monitoring contrast sensitivity before and after refractive surgery.

PURPOSE: To measure losses in contrast sensitivity at different orientations of grating stimuli in refractive surgery patients and show the importance of orientation when higher-order aberrations (HOAs) are altered. SETTING: Manchester Centre for Vision, Manchester Royal Eye Hospital, Manchester, United Kingdom. METHODS: Contrast sensitivity for sine-wave gratings of 12 cycles per degree was measured in 12 patients before and after different types of refractive surgery. Measurements were taken at 4 orientations with small (3.0 mm) and large (dilated) pupils using a binary search procedure. Higher-order aberrations were also measured. RESULTS: Changes in contrast sensitivity at different orientations were seen in patients before and after the surgery. All patients had significantly higher aberrations after surgery. Patients with increased amounts of postoperative HOAs were more likely to exhibit orientation effects. CONCLUSIONS: When contrast sensitivity is tested with gratings, it is advisable to measure more than 1 orientation. This particularly applies when root-mean-square error values are unusually high, such as in refractive surgery patients.

Clinical Characteristics and Outcomes of Moraxella Keratitis.

PURPOSE: Changing trends of infective keratitis have been identified worldwide. The aim of this study was to define the clinical associations, characteristics, and outcomes of patients with culture-proven Moraxella keratitis from a large tertiary corneal unit in the United Kingdom. METHODS: Patients with confirmed Moraxella isolates presenting between January 2004 and November 2016 were analyzed. Patient-related factors were examined, including patient demographics, date of presentation, clinical presentation, predisposing factors, best-corrected visual acuity (BCVA), treatment plans, and clinical outcomes. RESULTS: Eighty-six patients were identified, of whom 61 (70.9%) had at least one recognized predisposing factor. The median BCVA at presentation was 2.60 logarithm of the minimum angle of resolution (logMAR), which improved to median = 0.60 logMAR at final visit (P < 0.001). Visual improvement of more than 2 Snellen lines was achieved in 60.7% of eyes. A significant relationship (P = 0.003) between final vision and the presence of hypopyon was found, where for a fixed BCVA at presentation those with a hypopyon achieved a better final visual acuity of 0.69 logMAR. CONCLUSIONS: Our study showed that although most patients presented with at least one predisposing factor, some of the more traditionally perceived risk factors may need to be reconsidered. We have shown that with aggressive treatment, favorable visual outcomes can be achieved in most patients. Interestingly, the presence of hypopyon at presentation was linked to a better visual outcome, likely because such patients had more to gain in visual potential.

A fully automated cell segmentation and morphometric parameter system for quantifying corneal endothelial cell morphology.

BACKGROUND AND OBJECTIVE: Corneal endothelial cell abnormalities may be associated with a number of corneal and systemic diseases. Damage to the endothelial cells can significantly affect corneal transparency by altering hydration of the corneal stroma, which can lead to irreversible endothelial cell pathology requiring corneal transplantation. To date, quantitative analysis of endothelial cell abnormalities has been manually performed by ophthalmologists using time consuming and highly subjective semi-automatic tools, which require an operator interaction. We developed and applied a fully-automated and real-time system, termed the Corneal Endothelium Analysis System (CEAS) for the segmentation and computation of endothelial cells in images of the human cornea obtained by in vivo corneal confocal microscopy. METHODS: First, a Fast Fourier Transform (FFT) Band-pass filter is applied to reduce noise and enhance the image quality to make the cells more visible. Secondly, endothelial cell boundaries are detected using watershed transformations and Voronoi tessellations to accurately quantify the morphological parameters of the human corneal endothelial cells. The performance of the automated segmentation system was tested against manually traced ground-truth images based on a database consisting of 40 corneal confocal endothelial cell images in terms of segmentation accuracy and obtained clinical features. In addition, the robustness and efficiency of the proposed CEAS system were compared with manually obtained cell densities using a separate database of 40 images from controls (n = 11), obese subjects (n = 16) and patients with diabetes (n = 13). RESULTS: The Pearson correlation coefficient between automated and manual endothelial cell densities is 0.9 (p < 0.0001) and a Bland-Altman plot shows that 95% of the data are between the 2SD agreement lines. CONCLUSIONS: We demonstrate the effectiveness and robustness of the CEAS system, and the possibility of utilizing it in a real world clinical setting to enable rapid diagnosis and for patient follow-up, with an execution time of only 6 seconds per image.

Corneal clarity measurements in patients with keratoconus undergoing either penetrating or deep anterior lamellar keratoplasty.

PURPOSE: To compare the corneal clarity measurement between penetrating keratoplasty (PK) and deep anterior lamellar keratoplasty (DALK) in patients with keratoconus, using densitometry software for the Oculus Pentacam. METHODS: A retrospective comparative study was carried out at Manchester Royal Eye Hospital. Data were collected 12-18 months after corneal transplantation for keratoconus, including postoperative corneal densitometry, best corrected visual acuity (BCVA), central corneal thickness (CCT), and other relevant clinical details. RESULTS: Analysis of 37 keratoconus eyes from 36 patients found there was a significantly higher corneal densitometry measurement after DALK than PK. This was predominantly in the posterior layer of the concentric zone 0-2 mm of the cornea (P=0.0004). A significant correlation was found between postoperative BCVA and corneal densitometry in DALK groups at full thickness (P=0.03). This correlation was seen in the central 0-2 mm (P=0.03) and posterior 0-2 mm (P=0.04) zones. In addition, within the DALK group, a correlation was found between central corneal thickness and densitometry at full thickness 2-6 mm (P=0.007), central 0-2 (P=0.04), central 2-6 mm (P=0.01), and at posterior 2-6 mm (P=0.01) zones. CONCLUSION: This study showed that corneal densitometry measurement differs depending on the type of corneal transplantation used to treat keratoconus patients. Densitometry may have an important role to play in the final BCVA achieved by patients undergoing corneal transplantation for keratoconus. Analysis of Oculus Pentacam images provides an objective evaluation to monitor the cornea status after the surgery.

Association Between Season, Temperature and Causative Organism in Microbial Keratitis in the UK.

PURPOSE: Microbial keratitis (MK) is a major cause of corneal blindness worldwide. Variations in season and temperature can affect MK incidence due to specific causative organisms; however, few studies have examined these factors in the UK. METHODS: Retrospective review of all corneal scrapes from patients with MK presenting to Manchester Royal Eye Hospital, UK, between January 2004 and December 2015. Manchester's monthly temperature data were obtained from Met Office UK. Analysis was performed using logistic regression. RESULTS: From 4229 corneal scrapes, 1539 organisms grew (90.6% bacteria, 7.1% fungi, and 2.3% Acanthamoebae sp.). Gram-positive bacteria grew with increasing temperature [odds ratio (OR) 1.62, 95% CI: 1.11-2.39, P = 0.014], and fungi grew with decreasing temperature (OR 0.29, 95% CI: 0.16-0.51, P < 0.001). Moraxella sp. grew with decreasing temperature (OR 0.91, 95% CI: 0.86-0.96, P = 0.001). Compared with winter, overall culture positivity was significantly less likely in summer (OR 0.57, 95% CI: 0.38-0.87, P = 0.008) and spring (OR 0.65, 95% CI: 0.43-0.99, P = 0.045). Gram-negative bacteria were more likely in summer (OR 1.48, 95% CI: 1.06-2.09, P = 0.022) and autumn (OR 1.75, 95% CI: 1.24-2.47, P = 0.001). Candida sp. were less likely in summer (OR 0.25, 95% CI: 0.07-0.82, P = 0.027) and autumn (OR 0.18, 95% CI: 0.05-0.62, P = 0.009), and Acanthamoeba sp. were less likely in summer (OR 0.39, 95% CI: 0.15-0.92, P = 0.037) and spring (OR 0.26, 95% CI: 0.08-0.69, P = 0.011). CONCLUSIONS: Herein we report variation in the incidence of MK-causing organisms by season and temperature; this finding may aid clinicians in predicting possible causative organisms for MK at differing times of the year.

Corneal imaging and densitometry measurements in keratoconus patients to monitor disease progression and treatment outcomes after contact lens or Intacs treatment.

AIM: The aim of this study was to compare the pre- and posttreatment corneal densitometry and corneal thickness value of keratoconus (KCN) patients managed via contact lenses (CLs) or by both intrastromal corneal rings and contact lenses. PATIENTS AND METHODS: This prospective study was performed at the Manchester Royal Eye Hospital, UK. Patients were recruited before treatment and followed up for 12 months. Data of corneal densitometry and corneal thickness were collected using the Oculus Pentacam at the pretreatment visit and posttreatment visit at 12 months. RESULTS: Corneal clarity significantly differs between both groups at pre treatment at zone 0-2 mm for the anterior layer (P=0.002). The same diversity is present at zone 2-6 mm for the anterior layer (P=0.003) and posterior layer (P=0.008). The corneal clarity diversity found was not statistically significant at 12 months post treatment (P>0.05). Corneal thickness was found to be statistically significantly different between pre treatment and post treatment for the CL group for central corneal thickness (CCT) and thinnest area (P=0.01 and P=0.02), respectively. DISCUSSION: This study shows that KCN management with Intacs was found to be effective in maintaining corneal clarity for a longer time than that with CL alone. On the other hand, corneal clarity reduces with disease progression in cases managed with CLs only. Analysis of Oculus Pentacam images provides an objective evaluation to monitor the corneal status after these different pathways of management.

Subbasal nerve regeneration after LASEK measured by confocal microscopy.

PURPOSE: To determine the relationship between the degree of subbasal nerve regeneration and corneal sensation and tear film functions after LASEK using corneal confocal microscopy. METHODS: This prospective, observational, and longitudinal study included 35 LASEK patients who underwent tear breakup time assessment, phenol red thread test, corneal esthesiometry, and confocal microscopy preoperatively and 1, 3, and 6 months postoperatively. Subbasal nerve images were analyzed to assess nerve regeneration. RESULTS: Postoperatively, tear breakup time decreased significantly and had not returned to the preoperative level by 6 months after surgery. There were no significant differences in the phenol red thread test results before and after LASEK. Central corneal sensation decreased significantly 1 month after LASEK and returned to normal levels 3 months after surgery. Nerve fiber density, nerve branch density, and nerve fiber length and width decreased significantly after LASEK and had not returned to preoperative levels 6 months after surgery. Subbasal nerve fibers retained their vertical orientation after LASEK. There were no significant correlations between the length and density of subbasal nerve fibers and central corneal sensitivity, tear volume, or breakup time after surgery. CONCLUSIONS: Subbasal nerves were injured by LASEK and had not returned to preoperative levels 6 months after surgery. Corneal sensitivity was reduced after LASEK and returned to normal levels 3 months after surgery.

Subbasal nerve fiber regeneration after LASIK and LASEK assessed by noncontact esthesiometry and in vivo confocal microscopy: prospective study.

PURPOSE: To evaluate recovery of the corneal subbasal nerve plexus and corneal sensitivity after myopic laser in situ keratomileusis (LASIK) and laser-assisted subepithelial keratectomy (LASEK). SETTING: Manchester Centre for Vision, Royal Eye Hospital, Manchester, United Kingdom. METHODS: Thirty LASEK patients and 20 LASIK patients had slit-scanning confocal microscopy and noncontact corneal esthesiometry preoperatively and 1, 3, and 6 months after surgery. Images of the subbasal nerve plexus were analyzed using customized software to evaluate nerve regeneration. RESULTS: Central corneal sensitivity decreased significantly 1 month after LASEK and LASIK and returned to normal levels after 3 months. Corneal subbasal nerve fiber density, nerve branch density, nerve fiber length, and nerve fiber width decreased significantly 1 month after LASIK and had not returned to the preoperative levels by 6 months. Nerve fiber tortuosity decreased significantly 1 month after LASEK and returned to the preoperative levels 3 months after surgery. There were no significant differences in nerve fiber tortuosity before and after LASIK. Neither corneal sensitivity nor nerve fiber morphology was different between the 2 groups at any postoperative visit. CONCLUSIONS: Corneal sensitivity and subbasal nerve morphology were adversely affected by LASEK and LASIK. Corneal sensitivity recovered 3 months after the procedure, but subbasal nerves were still abnormal after 6 months. Despite the different forms of surgical trauma to corneal nerves with LASIK and LASEK, there was no apparent difference in the time course of recovery of corneal structure and function.

Subbasal nerve regeneration after penetrating keratoplasty.

PURPOSE: To evaluate subbasal nerve regeneration, corneal sensitivity, and tear film function after penetrating keratoplasty. METHODS: Twenty keratoplasty patients were assessed before and 1, 3, 6, and 12 months after penetrating keratoplasty by using noncontact corneal esthesiometry, tear breakup time measurement, the phenol red thread test, and confocal microscopy. Ten healthy control subjects were also assessed by using these techniques on 1 occasion. Subbasal nerve images were analyzed by using customized software to evaluate nerve regeneration. RESULTS: Before surgery, the subbasal nerve plexus could be imaged only in 11 patients because of the existence of pathology. Significant deficits in nerve morphology were apparent in these patients compared with control subjects. No subbasal nerves were detected over the 12-month postoperative period. Central corneal sensitivity decreased significantly after surgery and returned to near normal levels after 12 months. Tear breakup time was significantly shorter at 3 and 12 months after keratoplasty. There were no significant differences in the phenol red thread test results before and after surgery. CONCLUSIONS: There is no direct association between subbasal nerve fiber regeneration, central corneal sensitivity, and tear film stability and volume. The apparent recovery of corneal sensitivity to normal levels 12 months postoperatively, in the absence of clinically observable subbasal nerves, may be a methodologic phenomenon related to the inability of current-generation confocal microscopes to image fine regenerating nerves that mediate corneal sensibility.

Corneal clarity measurements in healthy volunteers across different age groups: Observational study.

The aim of this study was to standardize and investigate the changes in corneal clarity with age. Densitometry software for the Oculus Pentacam was used to examine corneal clarity at different age groups.A total of 192 eyes from 97 healthy participants were included in this cohort comparative nonrandomized, cross-sectional study. An Oculus Pentcam was used to image the cornea of healthy participants grouped by age (between 10 and 70 years old). Data from the densitometry output have been used to determine clarity in concentric zones and different depths of the cornea.Corneal densitometry (CD) across all ages showed significant differences between groups when divided into the following layers: anterior, central, and posterior or divided into 0 to 2, 2 to 6, and 6 to 10 mm concentric zones (P < .05). The most striking decrease in clarity occurred with age in all 3 layers of the periphery (6-10 mm) (P < .05). In addition, we showed that the 10 to 19-year age group had lower clarity than the 20 to 30-age group (P < .05), and after 30 years, the cornea shows a steady progression of increased or decreased clarity.The values for CD, as well as for separate subdivisions based on layer and surface area, might provide a standard for use in further studies and clinical practice. This study established that relation between CD and age is differed when the cornea is divided into layers and zones. This study suggests that there are other factors that may play an essential role in corneal clarity as well as age.

Evaluation of corneal symmetry after UV corneal crosslinking for keratoconus.

PURPOSE: The purpose of this study was to assess UV corneal crosslinking (CXL) treatment outcomes for keratoconus by evaluating the corneal regularity in patients through follow-up using the Oculus Pentacam. PATIENTS AND METHODS: A total of 18 eyes from CXL patients with keratoconus were studied before and after CXL treatment, and six eyes from six patients who were not treated with CXL served as controls. Treated patients had Pentacam images taken before CXL treatment and regularly 3 months post treatment up to the 12th month. Controls were imaged during their first appointment and after 12 months. Symmetry and asphericity were evaluated and correlated with both best-corrected visual acuity (BCVA) and maximum K-readings. RESULTS: In the CXL-treated group, there was a significant improvement in the index of symmetrical variation (ISV) and keratoconus index (KI) at 3 months and in the index of height asymmetry (IHA) and minimum radius of curvature (Rmin) at 9 months post treatment. On the contrary, the untreated group's indices showed some significant worsening in ISV, KI, central keratoconus index (CKI), and Rmin. A novel finding in our study was a slight positive shift of anterior asphericity in the 6 mm, 7 mm, and 8 mm 3 months after treatment, which had a correlation with BCVA (R2=0.390, p=0.053) and a strong correlation with maximum K-reading (R2=0.690, p=0.005). However, the untreated group had no significant changes after 1 year. CONCLUSION: The corneal asymmetrical shape is associated with the spherical aberration alteration influenced by temporal evolution of surface ablation and increased corneal haze. However, insignificant changes in symmetry attest the stabilization effect on cornea postoperatively as compared with controls.

Development of a three-dimensional organ culture model for corneal wound healing and corneal transplantation.

PURPOSE: To develop and evaluate a three-dimensional organ culture system of the cornea anterior chamber that could replicate the in vivo processes occurring during corneal wound healing and corneal transplantation. METHODS: Bovine corneoscleral buttons were clamped in a specially designed chamber through the sclera outside the limbus. The epithelium was exposed to air, and its anterior surface was automatically irrigated. The endothelial layer was perfused separately with media under normal intraocular pressure. Wound healing and corneal transplantation were observed using light, scanning, and transmission electron microscopy. RESULTS: The organ culture system maintained the epithelium, the putative epithelial stem cells in the limbus, the stroma, and the endothelium in good condition for the 10-day period during which the system was evaluated. The authors observed that the processes of wound healing and corneal transplantation in the model appeared similar to those occurring in vivo. CONCLUSIONS: In vitro model closely replicated the in vivo processes of wound healing and corneal transplantation. The authors believe this model will be useful for basic investigations into the cornea, such as study of the response of the cornea to surgery, wound healing, toxins, and therapeutic agents.