In this immunohistochemical study we applied a monoclonal antibody(mAb) to evaluate the expression pattern of lymphocyte functionassociated antigen 3(LFA-3) in rabbit`s corneas before and after intracorneal injection of Candida albicans. Ten right eyes were induced to get immunocompromized cornea with subconjunctival injection of 2mg of dexamethasone once a day for 3 days(group I), and 10 left eyes had normal cornea without subconjunctival injection of dexamethazone(group II). Each 2 corneas in both group I and II were resected at 3, 12, 24 and 72 hours after intracorneal injection of C. albicans. Each 2 corneas without intracorneal injection of C. albicans in both groups were used as a control. The results were as follows: LFA-3 was expressed weakly on corneal epithlium in control of group I and group II. Expression of LFA-3 on vascular endothelium of group II was somewhat stronger than that of group I, LFA-3 was expressed moderately on vascular endothelium, and was detected on corneal stroma at 3 hors after intracorneal injection in both groups. Expression of LFA-3 on corneal stroma was slightly increased in both group II, and markedly increased in group I at 12 hours after intracorneal injection. Group II showed slightly increased LFA-3 expression on corneal and II to be expressed on corneal endothelium and inflammatory cells at 24 hours after injection. Its expression on corneal epithelium, stroma and endothelium was more increased in group II than in group I at that time. Group I showed moderate LFA-3 expression on corneal epithelium, corneal endothelium and inflammatory cells, and strong expression on corneal stroma and vascular endothelium at 72 hours after infection. Otherwise, LFA-3 expression in group II was weak to moderate n corneal epithelium, corneal endothelium and inflammatory cell, and moderate on corneal stroma and vascular endothelium. In this study, it was found that expression of LFA-3 in group I was weaker than that in group II in control and at 3 hours after intracorneal injection of C. albicans, but group I showed more strong LFA-3 expression than group II after 12 hours of intracorneal injection.
Antigens, CD58 ; Candida albicans* ; Candida* ; Cornea ; Corneal Stroma ; Dexamethasone ; Endothelium ; Endothelium, Corneal ; Endothelium, Vascular ; Epithelium, Corneal ; Lymphocytes ; Rabbits*

In order to help define the possible role of adhesion molecules in corneal inflammation, we investigated the distribution of adhesion molecules in normal and Aspergillus fumigatus keratitis-induced corneas of rabbits in process if time. Each 4 corneas were resected at 3, 12, 24, and 72 hours after intracorneal injection with A. fumigatus. Normal corneas (4 eyes) were used as a control. Monoclonal antibodies to beta 1 subunit of VLAs, alpha 1 subunit of VLA-1, LFA-1, ICAM-1,and ELAM-1 were used for immunohistochemical staining of 20 corneas.The results were as follows: In normal cornea, beta 1 subunit of VLAs was expressed on all parts of the cornea, and ICAM-1 was expressed on corneal stroma and endothelium. Vascular endothelium showed expression of beta 1 subunit of VLAs and ICAM-1 after 12 hours of intracorneal injection, and alpha1 subunit of VLA-1 and ELAM-1 at 72 hours after injection. In inflamed cornea, beta 1 subunit of VLAs was expressed strongly at 72 hours after injection. Alpha1 subunit of VLA-1 was detected on corneal stroma after 12 hours of injection, and on corneal endothelium at 72hours after injection. Expression of beta 2 subunit of LFA-1 was weak on corneal stroma after 3 hours injection, and on corneal endothelium at 72 hours after injection. ICAM-1 expression was detected weakly on corneal epithelium, and increased on corneal stroma and endothelium at 72 hours after injection. ELAM-1 was expressed weakly on corneal stroma after 3 hours of injection, and on corneal endothelium at 72 hours after injection.In this study, it was found that the invasion of A. fumigatusinto the cornea causes localized inflammatory reaction that results in activation of corneal cells (keratocytes, corneal endothelial cells and epithelial cells), and subsequent expression of adhesion molecules in the cornea. Expression of adhesion molecules facilitates the inflammatory cells to be migrated into the cornea with inflammmation.
Antibodies, Monoclonal ; Aspergillus fumigatus* ; Aspergillus* ; Cornea* ; Corneal Stroma ; E-Selectin ; Endothelial Cells ; Endothelium ; Endothelium, Corneal ; Endothelium, Vascular ; Epithelium, Corneal ; Inflammation ; Integrin alpha1beta1 ; Intercellular Adhesion Molecule-1 ; Keratitis* ; Lymphocyte Function-Associated Antigen-1 ; Rabbits

PURPOSE: To analyze the features of corneal tissue in patients with posterior polymorphous corneal dystrophy (PPMD) using in vivo confocal microscopy (IVCM). CASE SUMMARY: Three patients with clinically diagnosed PPMD were examined using IVCM. Cross-sectioned corneal images of the corneal epithelium, Bowman's layer, stromal layer, Descemet's membrane, and endothelium were evaluated. IVCM demonstrated a depressed crater-like lesion, hyper-dense streak-like lesion, and surface irregularity of the corneal endothelium. Endothelial hypo-reflective vesicular and band-like lesions were also found. Pleomorphism and polymegathism were present with guttae and hyper-reflective endothelial nuclei. CONCLUSIONS: IVCM is a non-invasive and effective tool to diagnose PPMD.
Corneal Dystrophies, Hereditary ; Descemet Membrane ; Endothelium ; Endothelium, Corneal ; Epithelium, Corneal ; Humans ; Microscopy, Confocal

PURPOSE: Although a confocal microscopy has been largely used, there has been no domestic report concerning the normal structure of rabbit and human cornea by confocal microscope. In this study, we investigated confocal microscopic morphology of normal cornea of rabbit and human. MEHTODS: Using confocal microscope (ConfoScan 2.0, Fortune Technology, Italy), cross-sectioned corneal images of the corneal epithelium, Bowman's layer, stromal layer (anterior, middle, and posterior keratocyte), Descemet 's membrane, and endothelium were examined in vivo. RESULTS: In normal rabbit corneas, the superficial epithelium was polygonal and Bowman's layer was wrinkled. The stromal keratocyte nuclei was elliptical anteriorly and was with less cellularity posteriorly. The endothelium of rabbit was uniform, hexagonal in shape and had highly reflective cell borders. In human corneas, the superficial epithelium had highly reflective nuclei and regular wing cell or subepithelial nerve plexus was visible. Bowman's layer was fold-like structure. The shape of anterior and middle keratocyte nuclei were polygonal, but posterior keratocyte was oval shape. Also the nerve plexus of middle stromal layer was visible and the nucleus of endothelial cell was reflective hexagonal shape. CONCLUSIONS: The normal corneal morphological findings were studied by confocal microscopy. These data might provide the useful informations for the diagnosis of corneal disease.
Cornea ; Corneal Diseases ; Diagnosis ; Endothelial Cells ; Endothelium ; Epithelium ; Epithelium, Corneal ; Humans* ; Membranes ; Microscopy, Confocal

PURPOSE: To analyze the morphology and density of corneal tissue in patients with early stage Fuchs' corneal endothelial dystrophy (FCED) by in vivo confocal microscopy (IVCM). CASE SUMMARY: Each layer of the cornea in 2 patients with early-stage FCED was examined with IVCM (ConfoScan 4.0, NIDEK, Co. Ltd., albignasego, Italy). Cross-sectioned corneal images of the corneal epithelium, Bowman's layer, stromal layer, Descemet's membrane, and endothelium were evaluated. Corneal epithelium, Bowman's layer, and anterior stroma of both patients showed no abnormalities. Case 1 was diagnosed as Stage 1 FCED, demonstrating typical changes including pleomorphism, polymegathism, and the presence of guttae in the corneal endothelial layer. Case 2 was diagnosed as Stage 2 FCED, showing several hyper-reflective whitish dots in the posterior stroma, hypo-reflective vertical strands in the stroma adjacent to Descemet's membrane, and pleomorphism, polymegathism, and guttae in the corneal endothelial layer. CONCLUSIONS: IVCM is a non-invasive and effective tool to diagnose early-stage FCED.
Cornea ; Descemet Membrane ; Endothelium ; Epithelium, Corneal ; Humans ; Microscopy, Confocal

PURPOSE: To analyze the morphology and density of corneal tissue in patients with bullous keratopathy using in vivo confocal microscopy (IVCM). CASE SUMMARY: Four eyes of 4 patients with clinically diagnosed bullous keratopathy were examined using IVCM. Cross-sectioned corneal images of the corneal epithelium, Bowman's layer, stromal layer, Descemet's membrane, and endothelium were evaluated. In bullous keratopathy patients, the outline of corneal epithelial wing cells was blurred. Several hyper-reflective whitish dots were found at the corneal epithelial level and a reduction in the density and thickness of the corneal nerves was observed. There was also a reduction in the density of the stromal keratocytes. The outline of the stromal keratocytes was blurred. Some keratocytes resembled myofibroblast. The hypo-reflective vertical strands were observed in the stroma adjacent to the Descemet's membrane. Endothelium showed hyper-reflectivity with associated increased endothelial pleomorphism and polymegathism. CONCLUSIONS: In patients with bullous keratopathy, IVCM showed tissue edema in the corneal epithelium, stroma, and endothelium. Hyper-reflectivity appearances were noted due to the increase in the metabolic rate of the cells or extracellular matrix accumulation. Corneal endothelium showed morphological changes.
Descemet Membrane ; Edema ; Endothelium ; Endothelium, Corneal ; Epithelium, Corneal ; Extracellular Matrix ; Humans ; Microscopy, Confocal ; Myofibroblasts

PURPOSE: To evaluate the effect of reverse geometry lens (RGL) on correction of moderate degree myopia and cornea in patients of moderate-degree myopia. METHODS: 48 eyes of 82 eyes were chosen to wear reverse geometry lens and the remaining 34 eyes were fitted with rigid gas permeable (RGP) lens. The subjects were tested three days, one, two weeks, one, two and three months after fitting the lens. Uncorrected visual acuity, manifested refractive error, corneal thickness and corneal endothelial cell count were evaluated. corneal topography were performed. RESULTS: The uncorrected visual acuity was increased at 0.05 LogMAR after two weeks. and manifested refractive error decreased during the first months, thereafter stabilizing to -0.2 D (diopters). Corneal thickness was 540.4 micrometer and 583.0 micrometer in the center and mid-periphery, decreased to 505.2 micrometer and 572.7 micrometer, respectively, three months after RGL fitting (p<0.05). Endothelial cell count was 2663.5 cells/mm2 and 2541.5 cells/mm2 in the center and mid-periphery, 3 months after RGL fitting and endothelial cell count was not significantly different compared with that of pre-fitting. There was no statistical difference between RGL fitting group and RGP fitting group in the change of corneal endothelial cell count. CONCLUSIONS: RGL was found to be effective in correction of moderate-degree myopia. It had minimal effect on corneal endothelium, in short-term usage, however, the change of corneal thickness suggests that RGL fitting may effect on corneal epithelium and stroma.
Cornea* ; Corneal Topography ; Endothelial Cells ; Endothelium, Corneal ; Epithelium, Corneal ; Humans ; Myopia* ; Refractive Errors ; Visual Acuity

Laser-assisted in situ keratomileusis(LASIK) theoretically provide less corneal haze and more accurate correction. In the LASIK. however, photo refractive keratectomy(PRK) will be done on the stromal surface exposed after removal of the 130 micrometer thick lenticule. Deep stromal ablation after removal of 130 micrometer thick lenticule may result in endothelial damage. We observed endothelial cytoskeletal changes after deep PRK to evaluate the endothelial safety in rabbit. PRK was performed on the rabbit cornea without removing the epithelium with VISX 20/20 (160mj/cm2/pulse, 5Hz) to correct 6, 9, 12, and 15 diopters of myopia by single or multiple zone. Seven days after ablation, the rabbit was euthanized, corneas excised and endothelial F-actin was stained with NBD-phallacidin to demonstrate any cytoskeletal changes. No F-actin changes were observed in corneas after either single zone, 6 or 9 diopter correction, and multiple zone ablation. However, 12 diopters or higher correction in a single zone resulted in pleomorphism and polymegethism on focal areas. In the LASIK procedure, the endothelium might be damaged and further investigation for the endothelial preservation during the procedure is needed in human.
Actins* ; Cornea ; Endothelium ; Endothelium, Corneal ; Epithelium ; Humans ; Keratomileusis, Laser In Situ ; Myopia ; Photorefractive Keratectomy

PURPOSE: To analyze the morphology of corneal tissue in patients with Amiodarone-induced vortex keratopathy by in vivo confocal microscopy (IVCM). CASE SUMMARY: Four eyes of 2 patients with clinically diagnosed Amiodarone-induced vortex keratopathy were examined using corneal topography and IVCM. Cross-sectioned corneal images of the corneal epithelium, Bowman's layer, stromal layer, Descemet's membrane, and endothelium were evaluated. Location of corneal deposits examined by conventional slit-lamp microscopy was correlated with findings of corneal topography. The curvature map of corneal topography revealed an unusual irregular astigmatism with generalized mild steepening consistent with the location of the corneal deposits and the elevation map showed the change of corneal elevation according to the corneal deposits. Multiple hyper-reflective whitish dots were found at the corneal epithelial level and some were found at the anterior stromal level. Regarding the corneal endothelial layer, case 1 demonstrated normal corneal endothelial tissue, but case 2 showed several hyper-reflective whitish dots in the endothelium. CONCLUSIONS: In patients with Amiodarone-induced vortex keratopathy, IVCM showed corneal deposits in the corneal epithelium, stroma, and endothelium. Distribution of microdeposits in the corneal tissue caused an irregular astigmatism.
Astigmatism ; Corneal Topography ; Descemet Membrane ; Endothelium ; Epithelium, Corneal ; Humans ; Microscopy ; Microscopy, Confocal

PURPOSE: To report the confocal morphological changes of keratoconus in comparison with that of normal cornea. METHODS: Confocal microscopy (ConfoScan 2.0, Fortune Technology, Italy) was used to obtain data from healthy volunteers and keratoconus. We evaluated corneal morphological images of the corneal epithelium, Bowman's layer, stromal layer (anterior, middle, and posterior keratocyte), Descemet's membrane, and endothelium. RESULTS: In keratoconus cornea, the wing cells of epithelium had somewhat irregular margin, and the subepithelial nerve bundle was slightly enlarged. Near Bowman's membrane, highly reflective changes and tear like structure were visible. While the anterior stromal keratocyte had nuclei with a highly reflective density and indefinite cell border, posterior keratocyte had a more distinct shape with less cellularity. Near descemet's membrane, vertical fold (Vogt's striae) was observed and keratocytes with long processes arranged nearly in parallel. Morphological change was not observed in the endothelium. CONCLUSIONS: Our confocal microscopic findings of keratoconus may help early diagnosis and assessment of prognosis in the management of keratoconus.
Bowman Membrane ; Cornea ; Descemet Membrane ; Early Diagnosis ; Endothelium ; Epithelium ; Epithelium, Corneal ; Healthy Volunteers ; Keratoconus* ; Microscopy, Confocal ; Prognosis