• Journal of Korean Ophthalmic Optics Society
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• v.10 no.1
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• pp.63-69
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• 2005

The effects of R products of B company and natural disinfectant naringin, well-known chemical disinfectant on Corneal endothelium and epithelium of rabbit were observed by scanning electron microscope. The main component of naringin is extract of grapefruit seed, which is one of the flavonoid widely recognized as natural antioxidants and used as preservative agent of food and cosmetics. The chemical disinfectants cytotoxicity in cell cultures were announced by MIT assay and LDH leakage assay. But there has been no study about that chemical disinfectants and natural disinfectants is inserted drops on rabbit's cornea directly. In this test R products B company and natural disinfectants has been dropped on the cornea of rabbits for one weeks and observed cytotoxiciy by light camera after rose bengal staining. Also, we analysed the damage of corneal epithelium and endothelium morphologically after enucleating the cornea by scanning electron microscope.

• Proceedings of the Korean Society of Veterinary Clinics Conference
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• 2008.10a
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• pp.174-174
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• 2008

• 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.

• Journal of Pharmaceutical Investigation
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• v.24 no.1
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• pp.1-9
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• 1994

In order to study systemic peptide delivery through the ocular route, the stabilities of methionine enkephalin (Met-Enk) and $[D-ala^2]-methionine$ enkephalinamide (YAGFM) in the corneal extracts of rabbits were investigated using reversed phase HPLC. Met-Enk was found to be hydrolyzed most rapidly in the corneal epithelium, but YAGFM was relatively stable. Aminopeptidases appeared to contribute over 60% to the degradation of Met-Enk and the degradation rate of Met-Enk followed the first order kinetics. The half-lives of Met-Enk in the extracts of the corneal epithelium and endothelium were 36 and 673 min, respectively. From the effects of enzyme inhibitors, it was found that the application of the mixture of amastatin, thimerosal and EDTA was very useful for the inhibition of peptide degradation.

• Polymer(Korea)
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• v.38 no.6
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• pp.702-707
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• 2014

Corneal endothelium is mono-inner cell layer of cornea and lay on Descmet's membrane which comprised of various proteins called extracellular matrix such as fibronectin, collagen, laminin, and proteoglycan, etc. In this study, we fabricated transparent poly(lactic-co-glycolic acid) (PLGA) film because PLGA is widely used for tissue engineering based on their properties. We investigated the behaviors of rabbit corneal endothelial cells (rCEnCs) on PLGA film surfaces coated with various cell-adhesive molecules like fibronectin, laminin, collagen type I and IV and FNC coating mix. The morphologic images, proliferation and adhesion assay, immunofluorescence for ZO-1 and $Na^+/K^+-ATPase$ and RT-PCR for expression of specific markers were conducted. These results showed that PLGA film plays a role as CEnC carriers in vitro and the cell-adhesive molecules give positive effects on the behaviors of rCEnC.

• 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.

• 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.

• Journal of Korean Ophthalmic Optics Society
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• v.12 no.3
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• pp.19-25
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• 2007

It was studied how using soft contact lens multi-purpose solution (MPS), often used for medical treatment, effects the inhibition on cell growth, and how using the MPS demages eye cells, on rabbit eye's corneal epithelium and endothelium tissue. $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea) were used as MPS. After culturing Clone 1-5C-4 cell lines (Human conjunctival cell lines), cell growth inhibition rate was measured by MIT assay. By making Hematoxylin and Eosin stain specimen, the morphology was observed by optical microscope. In the In vivo experiment, 9 white rabbit eyes (18 eyes) were classified into 3 groups. The experiment group is left eyes (9 eyes) of rabbit, and MPS were dropped; however the control group, the right eyes (9 eyes), were only used a saline solution without a preservatives. After the dropping within the period, the cornea surface of rabbit endothelium tissue was observed using scanning electron microscopy (SEM).

• Journal of Biomedical Engineering Research
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• v.27 no.3
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• pp.125-130
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• 2006

The number of the living endothelial cells and the shape of those are very import clinical parameters for the evaluation of the quality of cornea. In this paper, we developed the automated endothelial cell counting and shape analysis algorithm for a confocal microscope. Since, the endothelial images from the confocal microscope has a non-uniform illumination and low contrast between cell boundaries and cell bodies, it is very difficult to segment the cells from the endothelial images. To cope with these difficulties, we proposed the new two stage image processing algorithm. At first stage algorithm, we used a high-pass filter and histogram equalization to compensate the non-uniform brightness pattern and a morphological filter and a watershed method are applied to detect the boundary of cells. From this stage, we could count the number of cells in an endothelial image. At second stage algorithm, we used a Voronoi diagram method to classify the shape of cells. This cell shape analysis and the percent of hexagonal cells are very sensitive in detecting the early endothelium damage. To evaluate the performance of the proposed system, we p개cessed seven endothelial images obtained using a confocal microscope. The proposed system correctly counted 95.5% cells and classified 92.0% of hexagonal cell shapes. This result is better than any others in this research area.

• Applied Microscopy
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• v.36 no.3
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• pp.141-153
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• 2006

The purpose of this study is to find out how soft contact lens multi-purpose solution (MPS), often used for medical treatment, effects the inhibition on cell growth and research the result of using MPS, suspected to demage eye cells, on rabbit eye's corneal epithelium and endothelium tissue. In this treatise, $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea) had been selected among the MPS. After culturing L929 roil line, cell growth inhibition rate was measured by MTT assay, and by making Hematoxylin and Eosin stain specimen. the morphology was observed by optical microscope. In the In vivo experiment,9 white rabbit eyes (18 eyes) were classified into 3 groups. The experimental group is left eyes (9 eyes) of rabbit, and MPS were dropped; however. the control group, the right eyes (9 eyes), were only used a saline solution without preservatives. After the dropping within the period, the cornea surface of rabbit eyes were stained by Rose bengal and observed. To figure out the changes of the corneal epithelium and endothelium tissue scanning electron microscopy (SEM) has been used. As the result, the rate of cell growth inhibition was 54%. 73% and 36%, respectively. Morphological changes represented that the shape has been changed into oval or round shape and those are not considered as a common formation of L929 cell line. When it comes to staining Rose bengal, each experiment group was stained red which is not shown in controls. The polygonal mosaic pattern of a corneal epithelium was disturbed in the picture taken by SEM; furthermore, the shape of the corneal endothelium was irregular. In conclusion, as we consider antimicrobial effect and the safety on living cells, it is necessary that we should improve concentration of preservatives and study continuously to develop a new preservatives without a toxic effect on the cornea surface.