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.

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.

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.

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.

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.

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.

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.

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.

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 Cross-Linking for Treatment of Fungal Keratitis: Using Confocal Laser Scanning Microscopy on an Ex Vivo Human Corneal Model.

Purpose: Some previous reports have established the use of photoactivated chromophore-induced corneal cross-linking (PACK-CXL) in treating fungal keratitis. The results of these case reports have often been conflicting. To systematically study the effect of PACK-CXL in the management of Fusarium keratitis, we have developed an ex vivo model of human corneal infection using eye-banked human corneas. Methods: Sixteen healthy ex vivo human corneas were divided into four study groups: (1) untreated control, (2) cross-linked, (3) infected with fungal spores, and (4) infected with fungal spores and then cross-linked. All infected corneas were inoculated with Fusarium oxysporum spores. The PACK-CXL procedure was performed 24 hours post inoculation for group 4. For PACK-CXL treatment, the corneas were debrided of epithelium; then 1% (wt/vol) isotonic riboflavin was applied dropwise at 5-minute intervals for 30 minutes and during the course of UV-A cross-linking for another 30 minutes. The corneas were imaged using a confocal microscope at 48 hours post inoculation, and the Fusarium hyphal volume and spore concentration were calculated. Results: The infected and then cross-linked group had a significantly lower volume of Fusarium hyphae, compared to the infected (P = 0.001) group. In the infected and then cross-linked group there was significant inhibition of Fusarium sporulation compared with the infected (P = 0.007) group. Conclusions: A model of human corneal infection was successfully developed for investigation of the effects of PACK-CXL on fungal keratitis. A treatment regimen of combined UV-A/riboflavin-induced corneal cross-linking appears to be a valuable approach to inhibit the growth and sporulation of Fusarium and suppress the progression of fungal keratitis.

Regeneration of Corneal Epithelium With Dental Pulp Stem Cells Using a Contact Lens Delivery System.

PURPOSE: The corneal epithelium is sloughed off surface of the eye by the action of blinking and is continually replaced by division and maturation of the limbal stem cells (LSCs). In the case of injury or disease, LSCs can be lost or damaged to a point at which the corneal epithelial layer is no longer maintained. leading to LSC deficiencies (LSCDs). When this occurs, the opaque conjunctiva overgrows the anterior surface of the eye, leading to vision impairment or loss. Dental pulp stem cells (DPSCs) are promising candidates as autologous LSC substitutes. In this study, contact lenses (CLs) are used as a novel medical device to deliver DPSCs onto corneal surface to enhance corneal epithelium regeneration. METHODS: Dental pulp stem cells labeled with green fluorescent Qtracker 525 were seeded onto the pretreated CLs, allowed to adhere, then delivered to debrided human corneas. Expression of KRT3, 12, 13, and 19 was investigated by immunostaining, then standard and confocal microscopy. RESULTS: Dental pulp stem cells were successfully isolated, labeled, and delivered to the corneal surface using CLs. Following removal of CLs, confocal microscopy showed that the DPSCs had migrated onto the cornea. Coexpression of KRT12 and green fluorescent Qtracker 525 confirmed that the DPSCs had transdifferentiated into corneal epithelial progenitors. Delimitation of KRT 19 and green fluorescence provides evidence that Qtracker 525-labeled DPSCs establish a barrier to the invasion of the cornea by conjunctiva. CONCLUSIONS: In this study we show that DPSCs, delivered using CLs, can be used to enhance repair and regeneration of the human corneal epithelium.

A fully automatic nerve segmentation and morphometric parameter quantification system for early diagnosis of diabetic neuropathy in corneal images.

Diabetic Peripheral Neuropathy (DPN) is one of the most common types of diabetes that can affect the cornea. An accurate analysis of the nerve structures can assist the early diagnosis of this disease. This paper proposes a robust, fast and fully automatic nerve segmentation and morphometric parameter quantification system for corneal confocal microscope images. The segmentation part consists of three main steps. First, a preprocessing step is applied to enhance the visibility of the nerves and remove noise using anisotropic diffusion filtering, specifically a Coherence filter followed by Gaussian filtering. Second, morphological operations are applied to remove unwanted objects in the input image such as epithelial cells and small nerve segments. Finally, an edge detection step is applied to detect all the nerves in the input image. In this step, an efficient algorithm for connecting discontinuous nerves is proposed. In the morphometric parameters quantification part, a number of features are extracted, including thickness, tortuosity and length of nerve, which may be used for the early diagnosis of diabetic polyneuropathy and when planning Laser-Assisted in situ Keratomileusis (LASIK) or Photorefractive keratectomy (PRK). The performance of the proposed segmentation system is evaluated against manually traced ground-truth images based on a database consisting of 498 corneal sub-basal nerve images (238 are normal and 260 are abnormal). In addition, the robustness and efficiency of the proposed system in extracting morphometric features with clinical utility was evaluated in 919 images taken from healthy subjects and diabetic patients with and without neuropathy. We demonstrate rapid (13 seconds/image), robust and effective automated corneal nerve quantification. The proposed system will be deployed as a useful clinical tool to support the expertise of ophthalmologists and save the clinician time in a busy clinical setting.

In vivo confocal microscopic corneal images in health and disease with an emphasis on extracting features and visual signatures for corneal diseases: a review study.

There is an evolution in the demands of modern ophthalmology from descriptive findings to assessment of cellular-level changes by using in vivo confocal microscopy. Confocal microscopy, by producing greyscale images, enables a microstructural insight into the in vivo cornea in both health and disease, including epithelial changes, stromal degenerative or dystrophic diseases, endothelial pathologies and corneal deposits and infections. Ophthalmologists use acquired confocal corneal images to identify health and disease states and then to diagnose which type of disease is affecting the cornea. This paper presents the main features of the healthy confocal corneal layers and reviews the most common corneal diseases. It identifies the visual signatures of each disease in the affected layer and extracts the main features of this disease in terms of intensity, certain regular shapes with both their size and diffusion, and some specific region of interest. These features will lead towards the development of a complete automatic corneal diagnostic system that predicts abnormalities in the confocal corneal data sets.

Biomechanical Changes of Collagen Cross-Linking on Human Keratoconic Corneas Using Scanning Acoustic Microscopy.

PURPOSE: To assess the biomechanical changes of collagen cross-linking on keratoconic corneas in vitro. METHODS: Six keratoconic corneal buttons were included in this study. Each cornea was divided into two halves, where one half was cross-linked and the other half was treated with riboflavin only and served as control. The biomechanical changes of the corneal tissue were measured across the stroma using scanning acoustic microscopy (SAM). RESULTS: In the cross-linked corneas, there was a steady decrease in the magnitude of speed of sound from the anterior region through to the posterior regions of the stroma. The speed of sound was found to decrease slightly across the corneal thickness in the control corneas. The increase in speed of sound between the cross-linked and control corneas in the anterior region was by a factor of 1.039×. CONCLUSION: A higher speed of sound was detected in cross-linked keratoconic corneal tissue when compared with their controls, using SAM. This in vitro model can be used to compare to the cross-linking results obtained in vivo, as well as comparing the results obtained with different protocols.

Biomechanical changes after repeated collagen cross-linking on human corneas assessed in vitro using scanning acoustic microscopy.

PURPOSE: To explore the biomechanical changes induced by repeated cross-linking using scanning acoustic microscopy (SAM). METHODS: Thirty human corneas were divided into three groups. In group A, five corneas were cross-linked once. In group B, five corneas were cross-linked twice, 24 hours apart. In group C, five corneas were cross-linked three times, 24 hours apart. The contralateral controls in all groups had similar treatment but without UV-A. The speed of sound, which is directly proportional to the square root of the tissue's elastic modulus, was assessed using SAM. RESULTS: In group A, the speed of sound of the treated corneas was 1677.38 ± 10.70 ms(-1) anteriorly and 1603.90 ± 9.82 ms(-1) posteriorly, while it was 1595.23 ± 9.66 ms(-1) anteriorly and 1577.13 ± 8.16 ms(-1) posteriorly in the controls. In group B, the speed of sound of the treated corneas was 1746.33 ± 23.37 ms(-1) anteriorly and 1631.60 ± 18.92 ms(-1) posteriorly, while it was 1637.57 ± 22.15 ms(-1) anteriorly and 1612.30 ± 22.23 ms(-1) posteriorly in the controls. In group C, the speed of sound of the treated corneas was 1717.97 ± 18.92 ms(-1) anteriorly and 1616.62 ± 17.58 ms(-1) posteriorly, while it was 1628.69 ± 9.37 ms(-1) anteriorly and 1597.68 ± 11.97 ms(-1) posteriorly in the controls. The speed of sound in the anterior (200 × 200 µm) region between the cross-linked and control corneas in groups A, B, and C was increased by a factor of 1.051 (P = 0.005), 1.066 (P = 0.010), and 1.055 (P = 0.005) respectively. However, there was no significant difference among the cross-linked corneas in all groups (P = 0.067). CONCLUSIONS: A significant increase in speed of sound was found in all treated groups compared with the control group; however, the difference among the treated groups is not significant, suggesting no further cross-links are induced when collagen cross-linking treatment is repeated.

Biomechanical properties of human corneas following low- and high-intensity collagen cross-linking determined with scanning acoustic microscopy.

PURPOSE: To assess and compare changes in the biomechanical properties of the cornea following different corneal collagen cross-linking protocols using scanning acoustic microscopy (SAM). METHODS: Ten donor human corneal pairs were divided into two groups consisting of five corneal pairs in each group. In group A, five corneas were treated with low-fluence (370 nm, 3 mW/cm(2)) cross-linking (CXL) for 30 minutes. In group B, five corneas were treated with high-fluence (370 nm, 9 mW/cm(2)) CXL for 10 minutes. The contralateral control corneas in both groups had similar treatment but without ultraviolet A. The biomechanical properties of all corneas were tested using SAM. RESULTS: In group A, the mean speed of sound in the treated corneas was 1677.38 ± 10.70 ms(-1) anteriorly and 1603.90 ± 9.82 ms(-1) posteriorly, while it was 1595.23 ± 9.66 ms(-1) anteriorly and 1577.13 ± 8.16 ms(-1) posteriorly in the control corneas. In group B, the mean speed of sound of the treated corneas was 1665.06 ± 9.54 ms(-1) anteriorly and 1589.89 ± 9.73 ms(-1) posteriorly, while it was 1583.55 ± 8.22 ms(-1) anteriorly and 1565.46 ± 8.13 ms(-1) posteriorly in the untreated control corneas. The increase in stiffness between the cross-linked and control corneas in both groups was by a factor of 1.051×. CONCLUSIONS: SAM successfully detected changes in the corneal stiffness after application of collagen cross-linking. A higher speed-of-sound value was found in the treated corneas when compared with the controls. No significant difference was found in corneal stiffness between the corneas cross-linked with low- and high-intensity protocols.