OBJECTIVETo evaluate the feasibility of using cultured corneal endothelial cell (CECs) transplantation for cornea endothelium cell destruction with Gelatin membrane as the carrier in rabbits.

METHODSThe cultured CECs were labeled by Brdu and subcultured in vitro on glutaraldehyde-fixed Gelatin membranes and then the membranes were glued by alpha-cyanoacrylate alkyl to 7.00 mm autologous rabbit corneal bottons whose endothelium were mechanically removed previously. The buttons were sutured in place. With this method the right eyes of 21 rabbits were transplanted with CECs, and the right eyes of another 17 rabbits were transplanted with non-cells carrier as controls. The rabbits were bred and observed by slit microscopy and confocal microscopy at 1, 2, 4, 8, and 12 weeks after surgeries. Also, introcular pressure and corneal thickness were measured by Perkin's tonometer and ultrasonic pachymeter. After 12 weeks, all the animals were sacrificed and the grafts were examined by light microscopy and electronic microscopy.

RESULTSCECs grew well on the gelatine memberane, and formed confluent monolayers in 3-5 days; the cell density reached as high as 2700 cells/mm2. After 2 weeks of operation, all corneal buttons were edema and began to be opaque. The control eyes remained opaque throughout the observation period. In eyes with CECs transplanted, the grafts began to be clear and thin 4 weeks after operation. The cell density of grafts decreased along with time, and the mean cell density of CECs transplantation buttons was (2023.3 +/- 330.3) cells/mm2 12 weeks after operation. The transplanted cells were stained with the anti-Brdu monoclonal antibody.

CONCLUSIONIt is feasible to culture and translate CECs with the Gelatin membrane.

Animals ; Cells, Cultured ; Corneal Endothelial Cell Loss ; pathology ; therapy ; Endothelial Cells ; transplantation ; Endothelium, Corneal ; cytology ; Rabbits

The method of gene transfer into corneal endothelium was investigated to provide a foundation for the study of TGF-beta 1 gene transfer to inhibit corneal graft rejection. Two days after direct injection of pMAM TGF-beta 1 mediated by liposome into the anterior chamber of rabbits, one half of corneas were made into paraffin slides and the endothelial layer was carefully torn from the other half to make a single layer slide of endothelia. By means of immunohistochemical technique, the plasmid pMAM TGF-beta 1 expression product TGF-beta 1 in the endothelia was detected. Specific TGF-beta 1 expression was positive in the endothelia on both the paraffin slide and the single layer slide. The results showed that by direct injection into the anterior chamber, foreign plasmid DNA could be transferred into the endothelia and its expression was obtained. This may provide a foundation for further study on TGF-beta 1 participating in local induction of corneal immune tolerance.
Anterior Chamber ; Corneal Transplantation ; Endothelium, Corneal/*drug effects ; Endothelium, Corneal/pathology ; Gene Transfer Techniques ; Immune Tolerance ; Transforming Growth Factor beta/administration &

PURPOSE: This study was carried out to clarify the histological characteristics of the interface of the corneal stroma and Descemet's membrane of the human eye. METHODS: Nighteen donor eyes without corneal pathology were examined by scanning and transmission electron microscopy. The Descemet's membrane including the corneal endothelium was cheked for scanning electron microscopy. The junctional characteristics of the posterior corneal stroma and Descemet's membrane was examined by transmission electron microscopy. RESULTS: The scanning electron microscopy showed that collagen sheet faced each other at the right angle near the Descemet's membrane and penetrated the Descemet's membrane with the irregular arrangement. The transmission electron microscopy showed that the electron-dense collagen filaments extended to the posterior stroma from Descemet's membrane. The arrangement of electron-dense collagen filaments paralleled with the arrangement of the collagen fibrils of the posterior stroma. CONCLUSIONS: The interface of the corneal stroma and Descemet's membrane was composed of two-typed extracellular materials without the intercellular specificatons.
Collagen ; Corneal Stroma* ; Descemet Membrane* ; Endothelium, Corneal ; Humans ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Pathology ; Tissue Donors

To evaluate the inhibiting effect of Homoharringtonine (HHT) on the corneal haze after excimer laser photorefractive keratectomy (PRK) in rabbits. 18 healthy rabbits which underwent PRK were randomly divided into three groups (A, B and C). The refractive degree of ablation was - 10.0DS in each group. Group A was locally treated with a piece of filter paper soaked with 1 mg/mL HHT for 5 min, and then the entire cornea was repeatedly irrigated with balance solution; Group B was dropped with 0.1 mg/mL HHT after PRK for 3 months; Group C was the control group. Corneal haze, histopathology, response, ect. were investigated. The corneal haze was significantly less in group A, while the difference between group B and group C was insignificant. Keratocytes and fibrocytes in corneal stroma were more active up to 3 months in group B and group C. Intraoperative use of topical HHT can reduce corneal haze after PRK in rabbits.
Corneal Opacity/etiology ; Corneal Opacity/*prevention & control ; Endothelium, Corneal/pathology ; Harringtonines/*administration & dosage ; Myopia/*surgery ; Photorefractive Keratectomy/*adverse effects ; Random Allocation

We report the clinical features and the course of traumatic corneal endothelial rings by trauma. Fourteen eyes (of fourteen patients) with traumatic endothelial rings (twelve cases of BB shot injury), were enrolled in this study. With median follow-up interval of 50 weeks, initial and final best corrected visual acuity, presence of combined injuries such as gross hyphema, and time of disappearance of traumatic endothelial rings were recorded. And specular microscopic examination was performed. The duration of existence of traumatic endothelial rings was mean 4.6 days. On the specular microscopic examination, the count of corneal endothelial cells in the injured eye decreased by mean 16.8% (ranged from 1 to 56%) than that in the opposite unjnjured eye. The duration of existence of traumatic endothelial rings was 3.5 days in the group without combined angle recession and was 6.1 days in the group with combined angle recession. We suggest that the possibility of traumatic corneal endothelial rings and resultant endothelial cell loss and their clinical potential should be always kept in mind in ocular trauma, particularly BB shot injury.
Adolescent ; Adult ; Aged ; Child ; Cornea/*injuries ; Endothelium, Corneal/*pathology ; Eye Injuries/*pathology ; Human ; Male ; Retrospective Studies ; Wounds, Gunshot/pathology

Light microscopy and electron microscopic examination were carried out on the corneal buttons of two patients who required penetrating keratoplasty for treatment of corneal complication following the intraocular injection of silicone oil to repair recurrent retinal detachments in aphakic eyes. Light microscopic examination demonstrated increased cellularity and irregularity of collagen fibers of stromal layer, defect of endothelial cell layer and endothelial degeneration. Electron microscopy examination demonstrated marked decrease in endothelial cell population density, accompanied by flattening and thinning of the remaining cells and attenuation of cell borders. There were silicone droplets in the endothelial cell layer and collagenous layer posterior to endothelial layer. These findings are well correlated to clinical manifestation and are thought to be rather due to barrier effect of silicone oil than direct toxicity.
Adult ; Corneal Diseases/*chemically induced/pathology/surgery ; Descemet Membrane/ultrastructure ; Endothelium, Corneal/drug effects/surgery/ultrastructure ; Humans ; Keratoplasty, Penetrating ; Male ; Recurrence ; Retinal Detachment/surgery ; Silicone Oils/*adverse effects

The method of gene transfer into corneal endothelium was investigated to provide a foundation for the study of TGF-beta 1 gene transfer to inhibit corneal graft rejection. Two days after direct injection of pMAM TGF-beta 1 mediated by liposome into the anterior chamber of rabbits, one half of corneas were made into paraffin slides and the endothelial layer was carefully torn from the other half to make a single layer slide of endothelia. By means of immunohistochemical technique, the plasmid pMAM TGF-beta 1 expression product TGF-beta 1 in the endothelia was detected. Specific TGF-beta 1 expression was positive in the endothelia on both the paraffin slide and the single layer slide. The results showed that by direct injection into the anterior chamber, foreign plasmid DNA could be transferred into the endothelia and its expression was obtained. This may provide a foundation for further study on TGF-beta 1 participating in local induction of corneal immune tolerance.
Animals ; Anterior Chamber ; Corneal Transplantation ; Endothelium, Corneal ; drug effects ; pathology ; Gene Transfer Techniques ; Immune Tolerance ; Rabbits ; Transforming Growth Factor beta ; administration & dosage ; biosynthesis ; genetics

Normally the cornea has a water content varying between 76-78%, a state of relative dehydration maintained through its own metabolism by the active transport of water and ions across its limiting membrane, the epithelium and endothelium. If the metabolism is grossly disturbed or if the effectivity of the limiting membrane is impaired, the living cornea will swell by the absorption of the fluid. Corneal edema are developed due to trauma, inflammation, glaucoma, degeneration, and neuropathic and metabolic conditions. Essential corneal edema are encountered for which no cause can be found, the condition apparantly occuring without other ocular pathology. A 29 years old Korean lady has been found to have bilateral essential edema of the cornea.
Absorption ; Adult ; Biological Transport, Active ; Cornea ; Corneal Edema* ; Dehydration ; Edema ; Endothelium ; Epithelium ; Glaucoma ; Humans ; Inflammation ; Ions ; Membranes ; Metabolism ; Pathology ; Water

The aim of this study was to evaluate the cellular response and morphological changes of cells on the intraocular lens(IOL) implanted over a course of time and to identify the basic mechanism of IOL adaptation to tissue reaction in the implanted eye by comparing polymethylmethacrylate (PMMA) IOL with heparin surface modified PMMA IOL. ECCE using Healon was done in 36 eyes of 36 rabbits. A heparin surface modified IOL was implanted in 18 eyes (Group I), while PMMA IOL was implanted into another 18 eyes (Group II). Corneal thickness and endothelial cell density were measured for 3 months. Postoperatively, the eyes were enucleated, and a cytopathologic examination of the cells on the surface of the IOL and their ultrastructural changes were observed with light and scanning microscope at various points of time. The findings of this present study suggested that heparin surface modified PMMA IOL reduced the degree of endothelial cell damage, postoperative tissue reaction, and pigment deposits on the surface of the IOL. These were statistically significant. The most important cell was considered to be the macrophage for the adaptation of IOL in the eye which gradually changedinto a fibroblast-like cell, giant cell and finally disappeared after forming an acellular membrane on the IOL.
Animals ; Cataract Extraction ; Cell Count ; Cornea/pathology ; Endothelium, Corneal/*pathology/ultrastructure ; *Heparin ; *Lenses, Intraocular ; Macrophages/pathology ; Materials Testing ; *Methylmethacrylates ; Microscopy, Electron, Scanning ; Rabbits ; Uvea/pathology/ultrastructure

PURPOSE: To analyze the effects of soft contact lenses on central corneal thickness and morphologic characteristics of the corneal endothelium in diabetic patients. MATERIALS AND METHODS: Ultrasound pachymetry and noncontact specular microscopy were performed on 26 diabetic patients who regularly use soft contact lenses (group 1), 27 diabetic patients who do not use soft contact lenses (group 2) and 30 normal subjects (group 3). We compared the values in each group using the Mann-Whitney test. RESULTS: The central cornea was found to be thicker in diabetic patients, both those who use and do not use contact lenses, than in the normal control group. The central corneal thickness was significantly higher in group 1 (564.73 +/- 35.41 microm) and group 2 (555.76 +/- 45.96 microm) than in the control group (534.05 +/- 27.02 microm), but there was no statistically significant difference between groups 1 and 2. Endothelial cell density was significantly different between the groups, and was smallest in the group of diabetic patients using contact lenses. The coefficient of variation of cell size was significantly higher and the percentage of hexagonal cells was significantly lower in contact lens using diabetic patients than in non-contact lens using diabetic patients and in the control group. CONCLUSION: Central corneal thickness and endothelial cell density is more affected by diabetes mellitus, and corneal endothelial cell morphology is more affected by contact lens use, when compared with normal subjects.
Adolescent ; Adult ; Case-Control Studies ; Contact Lenses, Hydrophilic/*adverse effects ; Cornea/pathology ; Corneal Endothelial Cell Loss/*etiology/pathology ; Diabetes Complications/*etiology/pathology ; Endothelium, Corneal/pathology ; Female ; Humans ; Male ; Statistics, Nonparametric ; Young Adult