• Journal of Veterinary Science
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• v.23 no.4
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• pp.62.1-62.10
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• 2022

Background: The corneal and limbal morphology relevant to corneal epithelial maintenance in ten different species was examined using histological methods. Objectives: The presence of a Bowman's layer, limbal epithelial cell, and superficial stromal morphology was examined in the following species to evaluate the differences in corneal thickness and epithelium: Java sparrows, frogs, macaws, spoonbills, red pandas, penguins, horses, Dobermans, orangutans, and humans. Methods: Corneal sections (4 ㎛) were obtained from ten ocular globes from three different animal classes: Aves, Amphibia, and Mammalia. All sections were stained with hematoxylin and eosin and periodic acid-Schiff reaction. After microscopy, all stained slides were photographed and analyzed. Results: Significant morphological differences in the corneal and limbal epithelia and their underlying stroma between species were observed. The number of corneal epithelial cell layers and the overall corneal epithelial thickness varied significantly among the species. The presence of a Bowman's layer was only observed in primates (orangutans and humans). Presumed supranuclear melanin caps were noted in four species (orangutans, macaws, red pandas, and horses) in the limbal basal epithelial layer (putative site of corneal epithelial stem cells). The melanin granules covered the apex of the cell nucleus. Conclusions: Supranuclear melanin capping has been described as a process within the epidermis to reduce the concentration of ultraviolet-induced DNA photoproducts. Similarly, there may be a relationship between limbal stem cell melanin capping as a protective mechanism against ultra-violet radiation.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.289-292
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• 2002

The corneal tissue consists of three layers : epithelium, stroma, and endothelium. Central cornea is a highly differentiated tissue whereas the limbus contains the epithelial stem cell. In the present study. we report the engineering of the three-dimensional reconstructed cornea derived from rabbit limbal epithelial and stromal cells. The differentiation degree of corneal stem cells were assessed in serum concentration and inoculation density of stromal cells. Optimal condition differentiation of corneal stem cells is achieved when 5% FBS was supplemented to culture medium and $1-2{\times}10^5$ cells/ml inoculation density of stromal cells.

• Applied Microscopy
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• v.28 no.4
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• pp.491-502
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• 1998

The present study was conducted to investigate the structural changes in rat cornea. Sixty eyes from one-day-old uneyed rats, fourty eyes from 4-weeks-old rats, and foully eyes from 10-weeks-old adult rats were used. With the increase of age, the epithelial layer was thickened by the addition of new successive cellular layers. Then, the new-born rat's epithelial cells formed a pentagonal shape, and the quality of decidual cells showed a high electron-density, although the boundary between cells was distinctive. The newly produced cells showed a low electron-density so that there was the distinctive difference between light and darkness. In Bowman's layer, collagen fibrils demonstrate a regularly arranged structure along with the age. In stroma's layer, the density of keratocytes was decreased and thereby Progressively flattened during the development. The collagenous layer of the adult rats was more distinctive than that of the new-born rats in a form of parallel alignment running vertically and horizontally.

• Journal of Korean Ophthalmic Optics Society
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• v.4 no.1
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• pp.1-6
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• 1999

The corneal epithelium is constantly shed and apoptosis may play an important role in this turn-over. We sought to define that serum-free medium was able to induce apoptosis of corneal epithelial cells. SV-40 transfected human corneal epithelial(HCE) cells were grown to 70% confluency in culture. Serum-free medium was added to cells and the cells incubated for 1, 2, 3, or 6 days. Apoptosis of cells at different times was assessed by staining cells with Giemsa or Hoechst 33342 and measuring DNA fragmentation using the TUNEL assay. HCE cells exposed to serum-free medium demonstrated a high incidence of apoptosis, which increased over time to $50{\pm}4%$ after 3 days. They also stained positively with TUNEL assay. Serum-free medium caused time dependent apoptosis of HCE cells. Thus, serum-like nutrient might be important in corneal epithelial cell homeostasis.

• Parasites, Hosts and Diseases
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• v.54 no.6
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• pp.697-702
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• 2016

Acanthamoeba keratitis has been increasing in recent years. Main risk factors are contact lens wear and their cleaning solutions. Most contact lens wearers use multipurpose disinfecting solutions (MPDS) for cleansing and disinfecting microorganisms because of its convenience. We determined amoebicidal effects of MPDS made in Korea and their cytotoxicity on human corneal epithelium cells. Fifteen commercial MPDS (A to O) were tested for their amoebicidal effects on Acanthamoeba castellanii trophozoites and cysts by using a most probable number (MPN) technique. Among them, 7 kinds of MPDS showed little or no amoebicidal effects for 24 hr exposure. Solutions A, B, G, H, L, and O showed positive amoebicidal effects, and solutions M and N killed almost all trophozoites and cysts after 24 hr exposure. However, 50%-N solution showed 56% cytotoxicity on human corneal epithelial cells within 4 hr exposure, and 50%-O solution also showed 62% cytotoxicity on human cells within 4 hr exposure. Solution A did not show any cytotoxicity on human cells. These results revealed that most MPDS made in Korea were ineffective to kill Acanthamoeba. The solutions having amoebicidal activity also showed high levels of cytotoxicity on human corneal epithelial cells. New formulations for improved MPDS that are amoebicidal but safe for host cells are needed to prevent Acanthamoeba keratitis.

• Journal of Korean Ophthalmic Optics Society
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• v.10 no.4
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• pp.283-292
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• 2005

This study investigated the effects of naringin, and chitosan in rabbits' corneas. Naringin, a glycone of naringenin, is a widely distributed bioflavonoid in the grapefruit and citrus peel, and it has already been reported as an antioxidant, antimicrobial, and anticancer agent. It has been used as a food preservatives and cosmetics. One of the natural preservatives, chitosan has also used in food preservatives, health drinks, and teas. Chitosan is distributed in the epithelium of crustacea, insects, and fungi. Naringin and chitosan have no harmful effects of cytotoxicity in the human body and they are recognized as an antibacterial for various forms of bacteria. The purpose of this study is to search for the ideal percentage of natural products to substitute the chemical preservatives occuring within the cornea and conjunctiva cytotoxicity and inflammations as wearing on soft contact lens. The present study compared the morphology of corneal epithelium and endothelium observed by light microscopy (LM) and transmission electron microscopy (TEM). In vivo methods, We investigated the effects of natural preservatives on soft contact lens. We inserted 3-4 drops of the naringin and chitosan, directly on rabbits' corneas 4 times per a day during one week. After enucleation of cornea, morphorgical damages of the epithelium and endothelium were observed by LM and TEM. In view of ultrastructure, chitosan caused siginficant damage on the epithelium and endothelium of cornea. The damage of cells was higher in chitosan treated cornea than 0.01, 0.1, and 1% of naringin. The 1% of naringin also expressed cell damage seriously. The results suggest that the most important thing is to use the reasonable percentage of preservatives for contact lens solutions.

• BMB Reports
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• v.46 no.2
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• pp.124-129
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• 2013

FK506 binding protein 12 (FK506BP) belongs to a family of immunophilins, and is involved in multiple biological processes. However, the function of FK506BP in corneal disease remains unclear. In this study, we examined the protective effects on dry eye disease in a Botulinum toxin A (BTX-A) induced mouse model, using a cell-permeable PEP-1-FK506BP protein. PEP-1-FK506BP efficiently transduced into human corneal epithelial cells in a time- and dose-dependent manner, and remained stable in the cells for 48 h. In addition, we demonstrated that topical application of PEP-1-FK506BP was transduced into mouse cornea and conjunctiva by immunohistochemistry. Furthermore, topical application of PEP-1-FK506BP to BTX-A-induced mouse model markedly inhibited expression levels of pro-inflammatory cytokines such as interleukin-$1{\beta}$ (IL-$1{\beta}$), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and macrophage inhibitory factor (MIF) in corneal and conjunctival epithelium. These results suggest PEP-1-FK506BP as a potential therapeutic agent for dry eye diseases.

• Journal of Veterinary Clinics
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• v.28 no.6
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• pp.562-565
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• 2011

A total of 29 eyes (25 dogs: one eye, 2 dogs: both eyes) with indolent corneal ulcer were treated with grid keratotomy from January 2008 to March 2010. The corneal lesions were reevaluated at 7-14 day intervals. The treatments had been repeated until fluorescein dye was not retained on the cornea and the epithelium did not appear to be loosely attached to the stromal layer. The healing rate of the corneal ulcers was 86.2%. The mean healing time ($mean{\pm}SD$) was $15.92{\pm}9.19$ days, ranged from 7 to 39 days. The lesions of remaining 4 eyes had deteriorated or not improved for more than 6 weeks. In those cases, $3^{rd}$ eyelid flap following grid keratotomy was applied. After 2 weeks, all of the eyes healed by the treatment. The results in this study suggest that grid keratotomy could be an excellent choice as an initial treatment for superficial corneal ulcers in dogs. In the cases of recurrence or to promote healing of the lesions, however, $3^{rd}$ eyelid flap following grid keratotomy is recommended.

• Journal of Korean Ophthalmic Optics Society
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• v.8 no.1
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• pp.65-71
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• 2003

The corneal epithelium is constantly being shed. The mechanism of corneal desquamation is not fully understood. Apoptosis, programmed cell death, may play a role. Apoptosis can be induced by a number of factors and different mechanisms. The study was performed to examine the apoptotic index induced in human corneal epithelial cells maintained in tissue culture by various apoptotic inducers. Various inducers, recombinant human cytokines($INF{\gamma}$, $TNF{\alpha}$, FASAb), actinomycin D. camptothecin, cycloheximide, dexamethasone and etoposide, were purchased from commercial suppliers. Inducers at manufacturer-recommended concentration were added to the corneal epithelial cells for 48 hours. Cell viability was measured using MTT assay. The cells were then assessed for the level of apoptosis. Morphologic changes and quantification of apoptotic cells were determined and counted under fluorescence microscope after inducers-treated human corneal epithelial (HCE) cells for 48 hours with Hoechst 33342 staining. Annexin V-FITC/PI staining and DePsipher assay. The expression of Fas protein was studied by immunocytochemistry. All inducers induced apoptosis in HCE cells in a dose dependent manner. Actinomycin D. camptothecin and etoposide induced apoptosis at lower than manufacturer-recommended concentration, while cytokines, cycloheximide and dexamethasone induced apoptosis at higher concentrations at the end of 48 hours. All inducers elicited typical apoptotic morphologic changes (chromatin condensation, nucleus fragmentations non-orange-red colored mitochondria) and expresses Fas protein highly. Apoptotic index of HCE cells by these inducers was different from the other cell lines. RNA synthesis inhibitor and topoisomerase inhibitors induced apoptosis at lower concentration than manufacturer-recommended concentration. Cytokines, cycloheximide and dexamethasone were able to produce apoptosis at 10 times higher concentrations. RNA synthesis inhibitor and topoisomerase inhibitors are more sensitive than intracellular receptor-activators in apoptotic induction of HCE cells.

• Journal of Korean Ophthalmic Optics Society
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• v.1 no.2
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• pp.19-35
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• 1996

This study was performed on the mouse to estimate the effect of UV radiation on the cornea in UV clean bench by LM & SEM. The results are as follows In the control groups, The cornea tissue have relatively compact and each layer have well identify, and the thick of cornea have constant. In the increasing experimental time, the experimental result have very different. The early experimental groups results have not severely degeneration. But, some substrate layer have a swelling and some epithelial tissue have not normal shape. The middle experimental groups results have very swelling of the stroma, the vacoule of some region the condensation of the epithelium, and the irregular arrangement of the endothelium. The last experimental groups results have shirinking of cornea tissue, the swelling and vacoule of the end endothelium, the partially disruption of epithelium, the irregular thick of the corneal tissue.