Keratocytes In Anterior Stroma Research Articles

Corneal Cellular and Neuroinflammatory Changes After SARS-CoV-2 Infection.

The purpose of this study was to evaluate corneal cellular and ultrastructural changes and to quantify the neuroinflammatory process in patients after mild severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Thirty patients after SARS-CoV-2 infection and 41 age-matched controls were examined. All subjects underwent in vivo confocal microscopy of the corneal cell layers and subbasal nerve fibers with the Heidelberg Retina Tomograph II. Semiautomated analysis of basal epithelial, anterior and posterior stromal keratocyte, and endothelial cell density was performed. Dendritic cell (DC) density and area were also calculated, and subbasal nerve plexus morphology was analyzed. The posterior stromal keratocyte density was significantly lower in patients after SARS-CoV-2 infection ( P = 0.0006). DC density in the central cornea was significantly higher in patients after SARS-CoV-2 infection ( P = 0.0004). There was a significant difference in the DC area between the 2 groups ( P < 0.0001). Significantly altered subbasal nerve fiber morphology was detected in patients after SARS-CoV-2 infection compared with healthy volunteers ( P < 0.05). Corneal cellular and ultrastructural changes demonstrated in this study suggest neuroinflammatory consequences of COVID-19 in the cornea in the absence of ophthalmoscopic alterations.

Clinical and microstructural changes with different iontophoresis-assisted corneal cross-linking methods for keratoconus.

To compare the clinical and microstructural changes induced by different transepithelial iontophoresis-assisted corneal cross-linking (I-CXL) methods for keratoconus. A total of 42 eyes of 42 patients with progressive keratoconus were divided into two groups. Group A received I-CXL for 5min, while group B received I-CXL for 10min. Visual acuity, optical coherence tomography (OCT), specular microscopy and confocal microscopy were evaluated preoperatively and at 1, 3, 6, and 12mo postoperatively. Twelve months after the operation, uncorrected visual acuity (UCVA) and corrected distance visual acuity (CDVA) were improved in both groups, with a better outcome in the I-CXL 10min group (P=0.025, 0.021, respectively). Kmax values decreased by 0.94±3.00 D in the I-CXL 10min group (P=0.033) but increased by 1.87±3.29 D in the I-CXL 5min group (P=0.012). OCT scans showed that the demarcation line was most visible and substantially deeper in the I-CXL 10min group. Confocal microscopy showed greater anterior stromal keratocyte decreases in the I-CXL 10min group than in the I-CXL 5min group at 3 and 6mo postoperatively (P<0.001); however, anterior stromal keratocytes and subbasal nerve density were not significantly different between the two groups at 12mo postoperatively. I-CXL for 10min more effectively halts the progression of keratoconus than I-CXL for 5min after 12mo of follow-up. However, long-term studies are needed to evaluate the efficacy and safety of I-CXL.

In vivo high resolution human corneal imaging using full-field optical coherence tomography.

We present the first full-field optical coherence tomography (FFOCT) device capable of in vivo imaging of the human cornea. We obtained images of the epithelial structures, Bowman's layer, sub-basal nerve plexus (SNP), anterior and posterior stromal keratocytes, stromal nerves, Descemet's membrane and endothelial cells with visible nuclei. Images were acquired with a high lateral resolution of 1.7 µm and relatively large field-of-view of 1.26 mm x 1.26 mm - a combination, which, to the best of our knowledge, has not been possible with other in vivo human eye imaging methods. The latter together with a contactless operation, make FFOCT a promising candidate for becoming a new tool in ophthalmic diagnostics.

Contact Lenses for Keratoconus- Current Practice.

Background:Keratoconus is a chronic, bilateral, usuallly asymmetrical, non-inflammatory, ectatic disorder, being characterized by progressive steepening, thinning and apical scarring of the cornea. Initially, the patient is asymptomatic, but the visual acuity gradually decreases, resulting in significant vision loss due to the development of irregular astigmatism, myopia, corneal thinning and scarring. The classic treatment of visual rehabilitation in keratoconus is based on spectacles and contact lenses (CLs).Objective:To summarize the types of CLs used in the treatment of keratoconus. This is literature review of several important published articles focusing on the visual rehabilitation in keratoconus with CLs.Method:Gas permeable (GP) CLs have been found to achieve better best corrected visual acuity than spectacles, eliminating 3rd-order coma root-mean-square (RMS) error, 3rd-order RMS, and higher-order RMS. However, they have implicated in reduction of corneal basal epithelial cell and anterior stromal keratocyte densities. Soft CLs seem to provide greater comfort and lower cost, but the low oxygen permeability (if the lens is not a silicone hydrogel), and the inability to mask moderate to severe irregular astigmatism are the main disadvantages of them. On the other hand, scleral CLs ensure stable platforms, which eliminate high-order aberrations and provide good centration and visual acuity. Their main disadvantages include the difficulties in application and removal of these lenses along with corneal flattening and swelling.Result:The modern hybrid CLs are indicated in cases of poor centration, poor stability or intolerance with GP lenses. Finally, piggyback CL systems effectively ameliorate visual acuity, but they have been related to corneal neovascularization and giant papillary conjunctivitis.Conclusion:CLs seem to rehabilitate visual performance, diminishing the power of the cylinder and the high-order aberrations. The final choice of CLs is based on their special features, the subsequent corneal changes and the patient’s needs.

Effects of contact lens wearing on keratoconus: a confocal microscopy observation.

To evaluate the corneal cell morphology of new keratoconus patients wearing two different types of rigid gas-permeable (RGP) contact lenses for 1y. Thirty nine eyes of 39 new keratoconus patients were selected and randomly fitted with two types of RGP contact lenses. Group 1 had 21 eyes with regular rigid gas-permeable (RRGP) contact lens and rest 18 eyes were in group 2 with specially designed rigid gas-permeable (SRGP) contact lens. Corneal cell morphology was evaluated using a slit scanning confocal microscope at no-lens wear and after 1y of contact lens wearing. After 1y of contact lens wearing in group 1, the mean anterior and posterior stromal keratocyte density were significantly less (P=0.006 and P=0.001, respectively) compared to no-lens wear. The mean cell area of anterior and posterior stromal keratocyte were also significantly different (P=0.005 and P=0.001) from no-lens wear. The anterior and posterior stromal haze increased by 18.74% and 23.81%, respectively after 1y of contact lens wearing. Whereas in group 2, statistically significant changes were observed only in cell density & area of anterior stroma (P=0.001 and P=0.001, respectively) after 1y. While, level of anterior and posterior stromal haze increased by 16.67% and 11.11% after 1y of contact lens wearing. Polymegathism and pleomorphism also increased after 1y of contact lens wearing in both the contact lens groups. Confocal microscopy observation shows the significant alterations in corneal cell morphology of keratoconic corneas wearing contact lenses especially in group 1. The type of contact lens must be carefully selected to minimize changes in corneal cell morphology.

Corneal Cell Morphology in Keratoconus: A Confocal Microscopic Observation.

To evaluate corneal cell morphology in patients with keratoconus using an in vivo slit scanning confocal microscope. A cross-sectional study was conducted to evaluate the corneal cell morphology of 47 keratoconus patients and 32 healthy eyes without any ocular disease. New keratoconus patients with different disease severities and without any other ocular co-morbidity were recruited from the ophthalmology department of a public hospital in Malaysia from June 2013 to May 2014. Corneal cell morphology was evaluated using an in vivo slit-scanning confocal microscope. Qualitative and quantitative data were analysed using a grading scale and the Nidek Advanced Visual Information System software, respectively. The corneal cell morphology of patients with keratoconus was significantly different from that of healthy eyes except in endothelial cell density (P = 0.072). In the keratoconus group, increased level of stromal haze, alterations such as the elongation of keratocyte nuclei and clustering of cells at the anterior stroma, and dark bands in the posterior stroma were observed with increased severity of the disease. The mean anterior and posterior stromal keratocyte densities and cell areas among the different stages of keratoconus were significantly different (P < 0.001 and P = 0.044, respectively). However, the changes observed in the endothelium were not significantly different (P > 0.05) among the three stages of keratoconus. Confocal microscopy observation showed significant changes in corneal cell morphology in keratoconic cornea from normal healthy cornea. Analysis also showed significant changes in different severities of keratoconus. Understanding the corneal cell morphology changes in keratoconus may help in the long-term monitoring and management of keratoconus.

In vivo corneal confocal microscopic analysis in patients with keratoconus.

To quantify corneal ultrastructure using laser scanning in vivo confocal microscopy (IVCM) in patients with keratoconus and control subjects. Unscarred corneas of 78 keratoconic subjects without a history of contact lens use and 36 age-matched control subjects were evaluated with slit-lamp examination (SLE), corneal topography and laser scanning IVCM. One eye was randomly chosen for analysis. Anterior and posterior stromal keratocyte, endothelial cell and basal epithelial cell densities and sub-basal nerve structure were evaluated. IVCM qualitatively demonstrated enlarged basal epithelial cells, structural changes in sub-basal and stromal nerve fibers, abnormal stromal keratocytes and keratocyte nuclei, and pleomorphism and enlargement of endothelial cells. Compared with control subjects, significant reductions in basal epithelial cell density (5817±306 cells/mm(2) vs 4802±508 cells/mm(2), P<0.001), anterior stromal keratocyte density (800±111 cells/mm(2) vs 555±115 cells/mm(2), P<0.001), posterior stromal keratocyte density (333±34 cells/mm(2) vs 270±47 cells/mm(2), P<0.001), endothelial cell density (2875±223 cells/mm(2) vs 2686±265 cells/mm(2), P<0.001), sub-basal nerve fiber density (31.2±8.4 nerves/mm(2) vs 18.1±9.2 nerves/mm(2), P<0.001), sub-basal nerve fiber length (21.4±3.4 mm/mm(2) vs 16.1±5.1 mm/mm(2), P<0.001), and sub-basal nerve branch density (median 50.0 (first quartile 31.2 - third quartile 68.7) nerve branches/mm(2) vs median 25.0 (first quartile 6.2 - third quartile 45.3) nerve branches/mm(2), P<0.001) were observed in patients with keratoconus. Significant microstructural abnormalities were identified in all corneal layers in the eyes of subjects with keratoconus using IVCM. This non-invasive in vivo technique provides an important means to define and follow progress of microstructural changes in patients with keratoconus.

Evaluation of Corneal Microstructure in Pseudoexfoliation Syndrome and Glaucoma: In Vivo Scanning Laser Confocal Microscopic Study

Purpose: To quantitatively evaluate corneas of patients with pseudoexfoliation syndrome (PXS) and pseudoexfoliation glaucoma (PXG) using in vivo scanning laser confocal microscopy (IVCM).Materials and Methods: The study population comprised 30 patients with PXS, 30 patients with PXG, and 30 normal control subjects. IVCM of the cornea was performed on all participants using the Rostock Cornea Module of the Heidelberg Retinal Tomograph (HRT). Mean outcome measures included density of basal epithelial cells, endothelial cells, and anterior and posterior keratocytes; and tortuosity and density of subbasal plexus nerves.Results: Mean densities of basal epithelial cells, endothelial cells, anterior and posterior keratocytes, and subbasal nerves differed significantly among the three groups. Subbasal nerve densities were significantly diminished in PXS and PXG patients (12.36 ± 2.89 and 6.8 ± 3.42 mm/mm2, respectively) compared with that of control subjects (16.13 ± 3.42 mm/mm2) (p < 0.05). Mean densities of anterior and posterior stromal keratocytes were significantly lower in PXS and PXG patients compared with control subjects (p < 0.05). Endothelial cell densities were 3073.63 ± 654.49, 2592.60 ± 276.36, and 2110.20 ± 620.53 cells/mm2 for control, PXS, and PXG groups, respectively (p < 0.05). The percentages of endothelial cell polymegathism and pleomorphism were higher in PXS and PXG patients compared with control subjects. Endothelial cell polymegathism and pleomorphism were more frequently associated with PXG.Conclusions: Results of this study demonstrate the existence of alterations in the (i) density of cells in the various layers of the cornea, (ii) cellular configuration of corneal endothelial cells, and (iii) density/diameter of the subbasal nerve plexus in patients with PXS, and that such alterations are common in patients with PXG. It would be beneficial to employ IVCM to assess the severity of pseudoexfoliation keratopathy (PXK).

Characteristics of the cornea in patients with pseudoexfoliation syndrome.

To quantify the morphological alterations in corneal nerve fibers and cells in patients with pseudoexfoliation syndrome (PEX) and their relationship with the presence of hyperreflective endothelial deposits observed using in vivo confocal microscopy. One eye each of 37 patients with PEX and 20 age-matched healthy control subjects was evaluated by in vivo corneal confocal microscopy. Patients with PEX were further classified into two groups: those with and without hyperreflective endothelial deposits. We evaluated the densities of basal epithelial cells, anterior and posterior stromal keratocytes, and endothelial cells and structure of sub-basal nerve fibers. The mean anterior and posterior stromal keratocyte and endothelial cell densities and corneal sub-basal nerve plexus variables were significantly lower in patients with PEX compared with those in healthy control subjects. The mean basal epithelial cell density did not significantly differ. Eyes with PEX presented decreased corneal sub-basal nerve plexus variables and cell densities in all corneas, except for the mean basal epithelial cell density. Further, a trend of lower corneal sub-basal nerve plexus measurements in patients with hyperreflective endothelial deposits compared with those without endothelial deposits was observed.

Corneal nerve fibre damage precedes diabetic retinopathy in patients with Type 2 diabetes mellitus

To quantify the morphological alterations in corneal nerve fibres and cells in patients with type 2 diabetes mellitus in relation to the severity of diabetic retinopathy. One hundred and thirty-two eyes of 132 patients with type 2 diabetes and 32 eyes of 32 healthy control subjects were evaluated with in vivo corneal confocal microscopy. Patients with diabetes were classified into three groups: patients without diabetic retinopathy, patients with non-proliferative diabetic retinopathy and patients with proliferative diabetic retinopathy. Anterior and posterior stromal keratocyte, endothelial cell and basal epithelial cell densities and sub-basal nerve fibre structure were evaluated. Significant reductions in basal epithelial cell, anterior stromal keratocyte and endothelial cell densities were observed only in patients with diabetic retinopathy. However, nerve fibre density, nerve branch density and nerve fibre length were reduced in patients without diabetic retinopathy and worsened progressively with increasing severity of retinopathy. Corneal cell pathology occurs in patients with diabetic retinopathy, but corneal nerve fibre damage seems to precede the development of diabetic retinopathy.

In Vivo Confocal Microscopy Analyses of Corneal Microstructural Changes in a Prospective Study of Collagen Cross-linking in Keratoconus

To use in vivo confocal microscopy (IVCM) to quantitatively analyze microstructural changes over time, after corneal collagen cross-linking for keratoconus. Prospective cohort study. A total of 38 eyes of 38 patients undergoing collagen cross-linking for keratoconus. Prospective, clinical cohort study of corneal collagen cross-linking in progressive keratoconus. Laser scanning IVCM performed preoperatively and at 1, 3, 6, and 12 months postoperatively. Density of corneal sub-basal nerves, anterior and posterior keratocytes, and corneal endothelium. Compared with baseline values, the mean sub-basal nerve density decreased significantly at 1, 3, and 6 months postoperatively (P < 0.01); however, this returned to preoperative values at 12 months (P = 0.57). One month postoperatively, there was complete absence of keratocyte nuclei in 86% of corneas. Anterior stromal edema with hyper-reflective cytoplasm and extracellular lacunae in a honeycomb-like appearance was observed and persisted at 3 months postoperatively. Scattered, presumed fragmented keratocyte nuclei, were observed at 1 and 3 months, but by 6 months, keratocyte repopulation of the anterior stroma was apparent. Quantitative analysis confirmed a significant decrease in the mean anterior keratocyte density 1, 3, and 6 months postoperatively (P ≤ 0.01) with return to baseline values at 12 months postoperatively (P = 0.57). The demarcation between treated and untreated corneal stroma appeared as a region where normal keratocytes transitioned into elongated, hyper-reflective, needle-like structures and then into large hyper-reflective stromal bands. There was no significant change in posterior keratocyte density or endothelial density at any postoperative time point. This prospective IVCM study revealed complete loss of the sub-basal nerve plexus and loss of anterior stromal keratocytes in the early postoperative period, with complete regeneration of the sub-basal nerve plexus and keratocyte repopulation by 12 months postoperatively. The posterior stroma and corneal endothelium were unaffected.

Evaluation of Contact Lens–Induced Changes in Keratoconic Corneas Using In Vivo Confocal Microscopy

To quantitatively analyze laser scanning in vivo confocal microscopy (IVCM) images of all corneal layers in contact lens-wearing and noncontact lens-wearing keratoconus patients. The study population included rigid gas permeable (RGP) contact lens-wearing keratoconus patients (group 1; N = 29), keratoconus patients who did not wear contact lenses (group 2; N = 30), and subjects who neither had keratoconus nor wore contact lenses (group 3; N = 30), with groups 2 and 3 matched to group 1 by age and sex. The central cornea was examined with IVCM in all subjects. The mean duration of contact lens wear was 5.50 ± 3.68 years (range, 2-15 years). Eyes with keratoconus showed significantly lower basal epithelial cell and anterior and posterior stromal keratocyte densities, as well as subbasal nerve fiber density, nerve branch density, and nerve fiber length compared with healthy control subjects. Furthermore, compared with group 2, group 1 had significantly lower basal epithelial cell density (4920 ± 476 cells/mm(2) vs. 4503 ± 461 cells/mm(2), P = 0.001) and anterior stromal keratocyte density (561 ± 91 cells/mm(2) vs. 464 ± 55 cells/mm(2), P < 0.001), but there was no significant difference for posterior stromal keratocyte density (P = 0.808), endothelial cell density (P = 0.699), or subbasal nerve fiber density (P = 0.142), nerve branch density (P = 0.614), and nerve fiber length (P = 0.850). Significant corneal microstructural abnormalities were observed in eyes with keratoconus. RGP contact lens wear was associated with a further reduction in the basal epithelial cell and anterior stromal keratocyte densities, but with no effect on posterior stromal keratocyte density, endothelial cell density, or corneal nerve morphology.

Evaluation of Corneal Microstructure in Keratoconus: A Confocal Microscopy Study

To compare the corneal microstructure in patients with manifest keratoconus (KCN), subclinical KCN, and topographically normal relatives of patients with KCN and in healthy controls. Prospective and cross-sectional study. We enrolled 145 subjects in the study. The participants were divided into 4 groups, based on clinical and topographical evaluation: the manifest KCN group (n = 30), the subclinical KCN group (n = 32), the KCN relatives group (n = 53), and the control group (n = 30). Corneal microstructure was assessed by corneal in vivo confocal microscopy in all of the individuals. Mean outcome measures were basal epithelial cell density, endothelial cell density, anterior keratocyte density, posterior keratocyte density, sub-basal nerve density, sub-basal nerve diameter, and stromal nerve diameter. The mean basal epithelial cell density, endothelial cell density, and sub-basal nerve diameter were not significantly different among the 4 groups (P = 0.057, P = 0.592, and P = 0.393, respectively). The mean anterior and posterior stromal keratocyte densities were significantly lower in the manifest group, in the subclinical group, and in the relatives group when compared with the control group (for both parameters; P < 0.001, P < 0.001, and P< 0.001, respectively). The mean stromal nerve diameter in the manifest group, subclinical group, and relatives group was significantly higher than in the control group (P = 0.001, P = 0.049, and P = 0.004, respectively). The anterior and posterior stromal keratocyte densities were statistically lower and stromal nerve diameter was statistically higher in patients with manifest KCN, subclinical KCN, and topographically normal KCN relatives compared with controls. Confocal microscopy may be useful for the determination of early corneal microstructural changes before manifestation of typical or subtle topographic signs.

LASIK operations with the Ziemer FEMTO LDV femtosecond laser flap creation after previous photorefractive keratometry or laser-assisted sub-epithelial keratomileusis

Editor, Enhancement procedures may be needed after previous refractive surgery in the cases of under or over correction and regression. Enhancement procedures can be performed by photorefractive keratometry (PRK) or laser-assisted in situ keratomileusis (LASIK). LASIK is an attractive option for retreatment after previous surface ablations such as PRK or laser-assisted sub-epithelial keratomileusis (LASEK). Main reasons for choosing LASIK are a quick recovery and the absence of haze. Previously, in LASIK, corneal flaps have been recreated using various microkeratomes (Comaish et al. 2002; Pitkänen et al. 2010). In recent years, femtosecond lasers have emerged as alternatives to the traditional microkeratome-based LASIK. Femtosecond laser technology has some advantages, such as less variation in flap thickness and a more uniform flap thickness throughout the whole flap, compared to microkeratome-based flaps. However, reports published on reoperations performed with the femtosecond laser technology after previous refractive surgery are still scarce and limited to Intralase (Tran et al. 2008). This study presents the outcomes of the first 89 consecutive reoperated eyes of 71 patients, previously treated with PRK (87 eyes) or LASEK (two eyes), which now underwent LASIK flap creation with the FEMTO LDV (Ziemer Ophthalmic Systems, Port, Switzerland) femtosecond laser and excimer laser treatment with the Allegretto Wave Concerto 500 Hz excimer laser (Wavelight AG, Erlangen, Germany). The patients were operated in the two Finnish Mehiläinen Hospitals, in Tampere and in Helsinki. In all the eyes operated, a superior hinge was used and the intended flap thickness was 90 μm. The diameter of the suction ring was chosen according to the preoperative keratometric K1 value measured before the primary operation. The study included 64 myopic eyes (residual spherical equivalent of refraction, SE, range −6.63 D to −0.25 D, mean SE −1.45 ± 1.01 D) and 25 hyperopic eyes (range SE +0.00 D to +2.88 D, mean SE +1.31 ± 0.91 D). Flap dimensions and the best-corrected visual acuities were measured and intra- and postoperative complications recorded. Mean flap thickness was 90.5 ± 5.4 μm (range, 74–104; Fig. 1) and mean flap diameter was 9.1 ± 0.2 mm (range, 8.8–9.7). Flap thickness was positively correlated with corneal thickness both in myopic (r = 0.33, P < 0.01) and in hyperopic previously surface-ablated eyes (r = 0.51, P < 0.01). The distribution of flap thickness in 89 previously surface-ablated eyes now reoperated with the FEMTO LDV using the intended flap thickness of 90 μm. Complications occurred in six eyes (5.9%). They all were observed in the previously PRK-treated eyes. Most of the complications were very mild, and none of them prevented further treatment with excimer laser. There were no changes in the Snellen lines of BSCVA in 73 eyes (94.8%), and one line was gained in four eyes (5.2%). The mean flap thickness in the previously surface-ablated eyes was equivalent to the one we obtained in our previous study evaluating 787 primary operations with the FEMTO LDV (Pietilä et al. 2010). In this study, flap thickness was affected by corneal thickness just as in our previous study (Pietilä et al. 2010). Although the reason for this positive correlation is unknown, it is, if anything, a clinically advantageous phenomenon. Corneal changes in the previously PRK- or LASEK-treated eyes are mainly ultrastructural (changes in keratocytes in anterior stroma, absence of Bowman’s membrane, subclinical haze in wound healing process). LASIK revision in these eyes poses potentially more risks, and the previously operated corneal tissue is likely to behave in a different way than in the primary LASIK procedure. It is, however, evident that LASIK revisions with the femtosecond technology can be safely performed in most of these cases. Although in general retreatment of previously operated eyes offers greater risks in refractive surgery, the FEMTO LDV femtosecond laser can be successfully used for corneal flap creation in previously surface-ablated eyes.

Mechanical Versus Alcohol-assisted Epithelial Debridement During Photorefractive Keratectomy: A Confocal Microscopic Clinical Trial

To compare the effects of mechanical versus alcohol-assisted epithelial debridement on corneal cellular elements after photorefractive keratectomy (PRK) using confocal microscopy. This randomized, clinical trial included 66 eyes from 33 patients with spherical equivalent refraction <-4.00 diopters (D). Mechanical versus alcohol-assisted epithelial debridement was performed during PRK. The right eye of each patient was randomly assigned to one group (mechanical group or alcohol-assisted group) and the fellow eye to the alternate group. Confocal examination was performed preoperatively and at 3 and 6 months postoperatively. Main outcome measures were keratocyte density and maximum anterior stromal light reflectivity. Mean epithelial healing time was 3.2 ± 0.4 and 3.0 ± 0.3 days in the mechanical and alcohol-assisted groups, respectively (P=.001). Anterior retroablation stromal keratocyte density was 704.3 ± 119.9 cells/mm² and 734.3 ± 103.7 cells/mm² at 3 months (P=.05) and 643.8 ± 134.4 cells/mm² and 696.7 ± 129.6 cells/mm² at 6 months (P=.02) in the mechanical and alcohol-assisted groups, respectively. No significant difference was noted in midstromal and posterior keratocyte density between the two groups. Maximum reflectivity was 61.56 ± 12.64 international units (IU) and 56.93 ± 7.86 IU in the mechanical and alcohol-assisted groups, respectively, 3 months after surgery (P=.018). Corresponding values were 49.46 ± 4.97 IU and 48.98 ± 4.60 IU, respectively, 6 months after surgery (P=.628). Due to more adverse effects of mechanical epithelial debridement on anterior keratocyte density and anterior stromal reflectivity, alcohol-assisted epithelial debridement is recommended as the procedure of choice for epithelial removal during PRK in patients with mild myopia.

Morphological characteristics of cornea in patients with vernal keratoconjunctivitis by in vivo laser scanning confocal microscopy

To explore the morphological characteristics on cornea in patients with vernal keratoconjunctivitis (VKC) by the application of in vivo laser scanning confocal microscopy (LSCM). The experimental design was retrospective observation case series (case control study). Twenty-six patients, each diagnosed as bilateral VKC, were enrolled in the study, among which 13 were tarsal form, 5 were bulbar form and the rest were mixed form. Nine patients had the clinical course less than one year, eight subjects longer than three years, and the rest between them. Another twenty-six healthy volunteers with matching age and gender were selected as normal control. All participants had their right eyes examined with the in vivo confocal microscopy (HRT II/RCM). Central cornea and superior peripheral cornea were chosen as the examination points. The images were recorded automatically and cellular density of each layer was analyzed by installed software. Software ImageJ was utilized to analyze the density, diameter, branch number and tortuosity of subbasal nerve fiber in VKC patients. Independent t test was performed to assess the differences on cellular density between VKC patients and normal control, as well as those between central and peripheral cornea in VKC patients. Fisher chi-square test was used to compare the infiltration rate of Langerhans cells in corneal epithelium between VKC patients and controls. ANOVA was applied to assess the differences in cellular density among three subtypes, as well as among different duration of VKC. Independent t-test and chi-square test were applied to analyze the parameters of subbasal nerve fiber. The morphological changes in cornea included the absence of superficial hyperreflective polygonal epithelial cells, infiltration of Langerhans cells in and(or) underneath corneal epithelium and activation of keratocytes in anterior stroma. Corneal epithelium conjunctivalization and stromal neovascularization could be identified in patients with corneal neovascular epithelium. Longitudinal or oblique dark striae could be found in the posterior stroma in patients with complicated keratoconus. The density of epithelial cells at central and peripheral cornea in healthy controls were (6033.1 ± 998.7) cells/mm(2) and (6098.4 ± 298.3) cells/mm(2), while that in VKC patients were (5972.2 ± 1148.2) cells/mm(2) and (6178.5 ± 318.9) cells/mm(2) respectively, the differences being no statistical significant between them (t = 1.191, 1.011; P = 0.238, 0.318). However, it's found in VKC patients that cellular density at peripheral cornea was significantly higher than that at central area (t = 2.249, P = 0.03). The density of anterior stromal cells at central and peripheral cornea in healthy controls was (1001.4 ± 125.3) cells/mm(2) and (924.6 ± 201.4) cells/mm(2), while that in VKC patients was (1184.5 ± 115.3) cells/mm(2) and (1101.4 ± 151.1) cells/mm(2), the difference bearing no statistical significance (t = 6.617, 3.439; P = 0.001). The density of posterior stromal cells in normal subjects and VKC patients was (537.7 ± 42.6) cells/mm(2) and (548.7 ± 79.8) cells/mm(2), that of endothelial cells was (2985.7 ± 401.2) cells/mm(2) and (3021.5 ± 383.3) cells/mm(2), respectively, neither difference had statistical significance (t = 0.174, 1.112; P = 0.864, 0.282). Langerhans cell infiltration could be identified in 61.5% (16 cases) VKC patients, which was significantly higher than normal control (2 cases, 7.7%) (χ(2) = 12.49, P = 0.001). Furthermore, much intense Langerhans cells infiltration was found in bulbar form and mix form than tarsal form. (t = 6.617, P = 0.001). The density and diameter of subbasal nerve fiber in VKC patients decreased significantly than those in normal subjects, whereas the tortuosity increased significantly. The morphological changes of cornea in VKC patients mainly involve corneal epithelium, subbasal nerve fiber and anterior stroma. In vivo LSCM is helpful in discriminating the subtypes of VKC.

Corneal Involvement in Crohn's Disease: An In Vivo Confocal Microscopy Study

To evaluate the corneal changes of patients with Crohn's disease (CD) using confocal microscopy and to investigate the association among confocal parameters and CD activity and CD treatment. Thirty consecutive patients (age: 42 ± 12 years; 19 women and 11 men) affected by CD and 30 control eyes (age matched and gender matched) underwent an ophthalmic examination and, in 1 eye chosen at random, confocal microscopy of the central cornea using the cornea module of Heidelberg Retina Tomograph. The following confocal parameters were evaluated: density of basal epithelial cells, epithelial dendritic cells, anterior and posterior stromal keratocytes, and endothelial cells; the subbasal plexus was assessed for number and tortuosity of the nerve fibers. Routine ophthalmic evaluation was normal in the whole population. At confocal microscopy, 40% of patients with CD had hyperreflective dots in the basal epithelium, which were absent in the control group. Activation of keratocytes was found in 86.6% of patients with CD and was absent in the control group. Compared with controls, patients with CD had lower density of dendritic cells (12.2 ± 26.3 vs. 50.3 ± 37.6 cells per square millimeter; P = 0.001). The other confocal parameters were similar in the 2 groups. No correlation between CD activity index and confocal changes was found. Confocal microscopy can detect subtle corneal changes in patients with CD, which may be signs of subclinical inflammation.

Topical 1% 5-fluorouracil in ocular surface squamous neoplasia: a long-term safety study

Background/aimsThe aim of this study was to evaluate the long-term corneal toxicity of topical chemotherapy with 1% 5-fluorouracil (5-FU) as a sole or adjuvant treatment of ocular surface squamous neoplasia...

Corneal Molecular and Cellular Biology Update for the Refractive Surgeon

To review clinically relevant progress in understanding cellular and molecular interactions in the cornea that relate to refractive surgical outcomes in patients. Recent published literature focused on femtosecond LASIK and surface ablation procedures, such as photorefractive keratectomy, was reviewed and correlated with clinical results of surgery. The femtosecond laser has a direct necrotic effect on stromal keratocytes, resulting in the release of cellular components that are chemotactic to bone marrow-derived inflammatory cells. Developments of the femtosecond laser led to lower energy delivery to the stroma and altered laser ablation profiles that decrease epithelial damage during the side-cut, and have markedly improved femtosecond LASIK to the point that the overall early postoperative healing response is indistinguishable from microkeratome LASIK. New studies have directly demonstrated the importance of surface irregularity and resulting structural and functional defects in the epithelial basement membrane, in the generation and persistence of anterior stromal myofibroblasts and haze following surface ablation procedures. These defects augment penetration of epithelium-derived TGF-beta, which is a critical modulator of myofibroblast development in the stroma. Studies on the mechanism of action of mitomycin C treatment to prevent haze have confirmed that the most powerful effect is on stromal cell proliferation and, therefore, decreased population of the anterior stroma with myofibroblast progenitor cells. An undesirable long-term effect of mitomycin C is diminished anterior stromal keratocyte density due to diminished keratocyte re-population. This raises concerns regarding future corneal anomalies in treated corneas. Basic research studies of refractive procedures provide important insights into the effects of wound healing on surgical outcomes.

Galα(1‐3)Gal expression of the cornea in vitro, in vivo and in xenotransplantation

To investigate alpha Gal (Gal alpha1-3Gal beta 1-4GlcNAc-R) expression of the porcine cornea in vitro, in vivo and after xenotransplantation. Using the GS-IB4 lectin (Griffonia simplifolia I isolectin B4), the expression of alpha Gal was evaluated in the normal porcine cornea and in cultured corneal stromal and endothelial cells of the second passages. The distribution of alpha Gal epitopes was also evaluated in corneal grafts at 4, 7 and 10 days after pig-to-rat orthotopic corneal transplantation. The expression of alpha Gal was mostly confined to the anterior stromal keratocytes of the normal porcine cornea. However, reactivity for GS-IB4 was markedly increased during cell culture passage (P)-by 18.0% (P1) and 39.0% (P2) in cultivated keratocytes, and by 62.1% (P1) and 87.1% (P2) in cultured endothelial cells. The expression of alpha Gal epitopes was gradually enhanced in all corneal layers of the graft after transplantation. Alpha Gal expression in the cornea was gradually induced during in vitro culture and after xenotransplantation, suggesting a role of putative Gal-related acute humoral rejection in porcine corneal xenotransplantation.