PURPOSE: We tried to determine the feasibility and efficiency of foreign gene transfer into corneal keratocytes using a hybrid EBV/retroviral vector as an investigative trial for gene therapy in corneal diseases. METHODS: LZRSpBMN-Z, alac Z-transducing hybrid EBV/retroviral vector, was transfected into Phoenix(T M) amphotropic packaging cells based on a 293T cell line and then collected without/with puromycin selection (puro (-)/puro (+) vector respectively). Cultured human and rabbit keratocytes were transduced with lac-Z gene using the puro (-) or puro (+) vector solutions, then stained with 5-bromo-4-chloro-3-indolyl galactopyranoside (X-gal). FACS-Gal analysis of transduced corneal keratocytes was also performed for calculating gene transfer efficiency. In addition, as an in vivo trial, we tried to transduce rabbit keratocytes by topical application of the vector supernatants following PRK or lamellar dissection of rabbit corneas. RESULTS: In vitro, both cultred human and rabbit keratocytes were transduced successfully with lac - Z gene. Transduction efficiency was 22% and 16% for human and rabbit keratocytes respectively with puro (-) vector, and slightly increased to 24% and 22% with puro (+) vector. In vivo corneas, however, no keratocytes were stained with X-gal. CONCLUSIONS: A hybrid EBV/retroviral vector, LZRSpBMN-Z, successfully transduced corneal keratocytes in in vitro conditions but not in vivo corneas.
Cell Line ; Cornea ; Corneal Diseases ; Corneal Keratocytes* ; Galactose ; Genetic Therapy ; Humans ; Product Packaging ; Puromycin

To describe the first case of the treatment of a corneoscleral cyst by distilled water injection into a corneal cyst. The anterior wall of a cyst of the limbal communication was punctured with a surgical blade. Aspiration and irrigation of the contents of the cyst with a 27-gauge anterior chamber cannula were performed repeatedly, three times. Distilled water, instead of balanced salt solution, was injected into the collapsed cyst, and was then aspirated completely after 5 minutes. The injection and aspiration of distilled water was repeated once more. The scleral cyst was surgically excised. Twelve months after surgery, several small white granular opacities, presumably epithelial cell nests, were observed on the interface of the collapsed cyst cavity, but there was no recurrence of the cyst. The best spectacle-corrected visual acuity (BSCVA) was 1.0 with a correction of +1.25-2.00 X 45. No significant change in central corneal endothelial cell density was noted. We suggest that this simple technique may represent an alternative method for the management of corneal cysts, and may have less risk of developing a corneal opacity or causing other serious damage to surrounding tissues.
Adolescent ; Cornea/*pathology ; Corneal Diseases/pathology/*therapy ; Corneal Topography ; Cysts/pathology/*therapy ; Human ; Injections ; Irrigation ; Male ; Scleral Diseases/pathology/*therapy ; Visual Acuity ; Water/*administration&dosage

PURPOSE: To evaluate the therapeutic effect of umbilical cord serum in the treatment of persistent epithelial defect of the cornea. METHODS: Fourteen eyes of 14 patients with persistent epithelial defect that had persisted for at least 2 weeks despite conventional treatment were treated with 20% umbilical cord serum eyedrops six times a day. The images of the epithelial defects were captured using a camera attached to a slit lamp biomicroscope and the areas of the epithelial defects were calculated. Treatment was considered effective for epithelial defect healing within 2 weeks, partially effective for healing within 2 to 4 weeks, and ineffective for healing requiring either more than 1 month or additional measures. RESULTS: Mean duration of epithelial defect before treatment was 7.2+/-6.3 weeks, and mean area was 7.86+/-7.32 mm2. Umbilical cord serum therapy was effective in 6 eyes (42.9%), partially effective in 6 (42.9%), and ineffective in 2 (14.2%). Nevertheless, the epithelial defects in both the ineffective eyes were eventually healed within 8 weeks. Mean healing time in effective or partially effective cases was 2.75+/-1.06 weeks. CONCLUSIONS: The use of umbilical cord serum eyedrops for the treatment of persistent epithelial defect is effective.
Treatment Outcome ; *Ophthalmic Solutions ; Middle Aged ; Male ; Humans ; *Fetal Blood ; Female ; *Epithelium, Corneal ; Corneal Diseases/*therapy ; Aged ; Adult

Descemet's membrane detachment (DMD) is an uncommon condition with a wide range of etiologies. More than likely, the most common cause is a localized detachment occurring after cataract surgery. We report three cases of Descemet's membrane detachment that occurred after uncomplicated phacoemulsification cataract surgeries. The first patient was managed without surgical intervention, the second patient was treated using an intracameral air injection, and the last patient was treated with an intracameral perfluoropropane (C3F8) gas injection. All three patients recovered their vision following the reattachment of Descemet's membrane. The three patients were treated according to the extent of the detachment.
Visual Acuity ; Middle Aged ; Male ; Humans ; Female ; Descemet Membrane/*pathology ; Corneal Diseases/*therapy ; Cataract Extraction/*adverse effects ; Aged

Amiodarone ; adverse effects ; Anti-Arrhythmia Agents ; adverse effects ; Arrhythmias, Cardiac ; drug therapy ; Corneal Diseases ; chemically induced ; Humans ; Male ; Middle Aged

In this paper, a case of corneal squamous cell carcinoma is reported. Invasive squamous cell carcinoma of the cornea is a rare disorder and has not been previously described in the Korean literature. In this case, the invasive squamous cell carcinoma of the cornea was treated by complete excision and cryotherapy. No evidence of metastasis or recurrence has been found since the procedure. Complete excision and adjunctive cryotherapy has become the treatment of choice because of the higher recurrence rate following a simple excision.
Aged ; Carcinoma, Squamous Cell/*pathology/surgery/therapy ; Case Report ; Corneal Diseases/*pathology/surgery/therapy ; Cryotherapy ; Eye Neoplasms/*pathology/surgery/therapy ; Human ; Male ; Neoplasm Invasiveness

The purpose of this study is to characterize and compare the ultrastructural changes occurring during the in vivo cultivation of corneal epithelium on amniotic membrane (AM) at several different time points. Corneal burn patients (n=7) with a corneal epithelial defect and severe limbal damage were selected. Initially, AM transplantation with limbal autograft was performed at the acute stage of corneal burn to reconstruct the damaged ocular surface. One to six (mean interval; 3.3+/-1.2) months later, the central part of AM containing an in vivo expanded corneal epithelium was excised and retransplanted in adjacent lesions. The excised epithelium with AM was examined by electron microscopy and immunohistochemical study. By electron microscopy, one and two months after expansion, cultivated epithelium on AM showed an undifferentiated epithelium and an incomplete basement membrane (BM). But, after three months, the cultivated epithelium began to differentiate into a multilayered epithelium with a continuous BM with increased hemidesmosomes. These findings were further confirmed by immunohistochemical study, that cytokeratin K3 was expressed in the cultivated corneal epithelium and newly formed BM was partially positive of collagen IV at three months. At least 3 months may be needed for the proliferation and differentiation of in vivo cultivated corneal epithelium on AM.
Stem Cells/cytology ; Stem Cell Transplantation/*methods ; Middle Aged ; Microscopy, Electron ; Male ; Keratin-3/biosynthesis ; Immunohistochemistry ; Humans ; Epithelium, Corneal/cytology/*metabolism/*pathology/*transplantation ; Corneal Diseases/*therapy ; Burns/*surgery/therapy ; Biological Dressings ; Amnion/*ultrastructure ; Adult

Topical fibronectin, autologous and homologous, was used to treat nine patients (eleven eyes) with persistent corneal epithelial defects and corneal ulcers that failed to improve with standard therapy. The fibronectin was purified from autologous and homologous plasma by gelatin-Sepharose 4B affinity chromatography and administered topically, 500 micrograms/ml five times a day, for three weeks. Complete or nearly complete reepithelialization was achieved in all patients regardless of the source of fibronectin, autologous or homologous. But healing times varied. The average healing time was 41.7 +/- 14.7 days (35.7 +/- 12.4 days for autologous, 50.8 +/-14.4 days for homologous). Ocular symptoms were relieved significantly, and no side effects were observed. Over an average follow-up period of 5.2 months, no recurrences were noted. The results showed that homologous, as well as autologous, fibronectin was effective in patients with persistent corneal epithelial defects and corneal ulcers.
Administration, Topical ; Adult ; Aged ; Blood Proteins/isolation & purification ; Chromatography, Affinity ; Corneal Diseases/*drug therapy ; Corneal Ulcer/*drug therapy ; Epithelium/drug effects ; Female ; Fibronectins/administration & dosage/isolation & purification/*therapeutic use ; Humans ; Male ; Middle Aged ; Visual Acuity ; Wound Healing/drug effects

PURPOSE: In Asian countries, laser iridotomy for the treatment of angle-closure glaucoma is a common cause of bullous keratopathy, which may be associated with a shallow anterior chamber and dark iris pigmentation in Asians. Several cases of corneal decompensation after argon laser iridotomy have been reported. In the present study, we evaluated the harmful effects of argon laser iridotomy on the corneal endothelium. METHODS: Argon laser iridotomy was performed on the right eyes of pigmented rabbits. Changes in corneal thickness and endothelial cell density after laser iridotomy were evaluated. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was performed for assessment of corneal endothelial cell apoptosis. Combined staining with alizarin red and trypan blue, as well as a live/dead cell assay, were performed for evaluation of damage to the corneal endothelium induced by laser iridotomy. RESULTS: Corneal thickness did not change immediately after laser iridotomy; however, a significant increase was observed 24 hours after iridotomy (p = 0.001). The endothelial cell density of laser-treated eyes four days after laser iridotomy was significantly decreased compared with control eyes (p < 0.001). TUNEL staining showed many TUNEL-positive cells in the corneal endothelium and corneal stroma. No endothelial trypan blue-stained cell nuclei were observed after laser iridotomy; however, several large endothelial cells with damaged membrane integrity were observed. The live/dead cell assay clearly showed a large number of dead cells stained red in several areas throughout the entire corneal button 24 hours after iridotomy. CONCLUSIONS: Argon laser iridotomy induces corneal endothelial cell apoptosis in pigmented rabbit eyes, resulting in decreased endothelial cell density.
Animals ; Apoptosis ; Corneal Diseases/pathology/*surgery ; Disease Models, Animal ; Endothelium, Corneal/*pathology ; In Situ Nick-End Labeling ; Iris/*surgery ; Laser Therapy/*methods ; Lasers, Gas/*therapeutic use ; Ophthalmologic Surgical Procedures/*methods ; Rabbits

PURPOSE: In Asian countries, laser iridotomy for the treatment of angle-closure glaucoma is a common cause of bullous keratopathy, which may be associated with a shallow anterior chamber and dark iris pigmentation in Asians. Several cases of corneal decompensation after argon laser iridotomy have been reported. In the present study, we evaluated the harmful effects of argon laser iridotomy on the corneal endothelium. METHODS: Argon laser iridotomy was performed on the right eyes of pigmented rabbits. Changes in corneal thickness and endothelial cell density after laser iridotomy were evaluated. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was performed for assessment of corneal endothelial cell apoptosis. Combined staining with alizarin red and trypan blue, as well as a live/dead cell assay, were performed for evaluation of damage to the corneal endothelium induced by laser iridotomy. RESULTS: Corneal thickness did not change immediately after laser iridotomy; however, a significant increase was observed 24 hours after iridotomy (p = 0.001). The endothelial cell density of laser-treated eyes four days after laser iridotomy was significantly decreased compared with control eyes (p < 0.001). TUNEL staining showed many TUNEL-positive cells in the corneal endothelium and corneal stroma. No endothelial trypan blue-stained cell nuclei were observed after laser iridotomy; however, several large endothelial cells with damaged membrane integrity were observed. The live/dead cell assay clearly showed a large number of dead cells stained red in several areas throughout the entire corneal button 24 hours after iridotomy. CONCLUSIONS: Argon laser iridotomy induces corneal endothelial cell apoptosis in pigmented rabbit eyes, resulting in decreased endothelial cell density.
Animals ; Apoptosis ; Corneal Diseases/pathology/*surgery ; Disease Models, Animal ; Endothelium, Corneal/*pathology ; In Situ Nick-End Labeling ; Iris/*surgery ; Laser Therapy/*methods ; Lasers, Gas/*therapeutic use ; Ophthalmologic Surgical Procedures/*methods ; Rabbits