• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.262-269
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• 2005

Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

• Korean Journal of Veterinary Research
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• v.56 no.3
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• pp.155-160
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• 2016

This study was conducted to evaluate three different mixed formulations of sodium hyaluronate (SH) and carboxymethyl cellulose (CMC) using a low-humidity air flow-induced rat dry eye model and determine the most suitable mixture. The total thickness of the cornea, corneal epithelial thickness, corneal stroma thickness, damaged corneal epithelium percentage region, thickness of the bulbar conjunctiva epithelium, number of goblet cells, goblet cell occupation percentage region, and damaged bulbar conjunctiva epithelium percentage region were measured by histomorphological evaluation. After 5 h exposure to drying airflow, the thickness of the cornea and conjunctiva was decreased with desquamation of the corneal and conjunctiva epithelium. However, these dry eye symptoms were markedly inhibited by treatment with the reference and test formulations. More favorable effects on decreased thickness were detected in response to the CMC than the SH. However, SH had a greater protective effect against corneal and conjunctiva epithelial damage. The application of a mixture of 0.1% SH and 0.2% CMC showed more favorable effects on the corneal and conjunctival damage and the stabilization of the ocular surface than SH or CMC alone.

• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.358-366
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• 2007

This study was performed to investigate the application of bovine amniotic membrane for canine corneal ulcer and to compare bovine amniotic membrane flap to nictitating membrane flap. Corneas were treated with 8.0mm trephine and 100% ethanol was applied and corneal epithelium was removed. This experiment was divided into three groups; control group, nictitating membrane flap group and amniotic membrane + nictitating membrane flap group. To compare corneal healing, fluorescein dye test, histopathological examination and TUNEL assay were performed. The healing size of the corneal epithelium in was largest among groups and it was significantly different with the other groups (P<0.05). Also, inflammatory cell deposits and keratocyte apoptosis in amniotic membrane flap group were smallest, but the width of the corneal epithelium was thickest among groups. Therefore, we tentatively predicted that the amniotic membrane flap would be an alternative effective method for the treatment of superficial canine corneal ulcer.

• Journal of Veterinary Clinics
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• v.35 no.6
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• pp.311-314
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• 2018

Squamous cell carcinoma (SCC) is a form of neoplasm that origins from the epithelial surface of many organs. Ocular occurrence in small animals is rare, especially in the central cornea without the involvement of limbus or conjunctiva. In the current case, a 10-year-old, spayed female Shih-Tzu was presented with a central corneal mass. Through an ophthalmic examination the sign of corneal scarring around the mass was found. Pink fleshy lesion that protruded outward was removed through superficial keratectomy. The mass with the size of $8.5{\times}6.5$ millimeter was histopathologically diagnosed as corneal SCC infiltrating not only the epithelium, but also the superficial corneal stroma with surrounding superficial lymphoplasmacytic and neutrophilic chronic keratitis. Also, the adjacent, non-affected corneal epithelium was markedly hyperplastic and keratinized; the adjacent stroma was moderately vascularized and fibrotic. The pulse-dose therapy using 1% topical 5-flourouracil were applied for five cycles. The tumor has not recurred through nine months of follow-up time.

• KSBB Journal
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• v.20 no.4
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• pp.305-310
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• 2005

Reconstruction of rabbit corneal epithelium was performed through dynamic culture method using self-manufactured amniotic membrane supporter and lyophilized amniotic membrane. Rabbit corneal epithelial cells were cultured and cryopreserved after isolation from limbus, and the cells could be proliferated by passage number 10. The basal layer was well formed, and the epithelium layer was constructed tightly by the increase of cell proliferation and differentiation by dynamic culture method than static culture. Thus, the reconstruction of the corneal epithelium using lyophilized amniotic membrane is considered to be a good in vitro model for transplantation of corneal epithelium to patients with a severely damaged cornea.

• 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.

• Journal of Veterinary Clinics
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• v.18 no.4
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• pp.418-423
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• 2001

To investigate the modulation of nerve growth factor (NGF) during corneal epithelial wound healing and the effect of topical NGF on corneal epithelial wound healing in dogs. An axial epithelial defect was created in the right eye using 6mm axial corneal mechanical debridement while the left served as an unwounded control. The tears were collected from both eyes during 1 week and the corneal epithelium was processed for the measurement of NGF at day 0 and 7. The NGF content of tears and corneal epithelium was determined by enzyme-linked immunosorbent assay. In another experiment, the animals were divided into 3 groups. The right eyes in each group were treated every six hours with 200 ug/ml of recombinant human (rh) NGF, murine NGF, or 600 ug/ml of anti-NGF blocking antibody. The left eye of each animal was treated with bovine serum albumin (BSA) to serve as controls. Wound healing was analyzed using NIH image software. Tear NGF was markedly increased in the wounded eyes, relative to tears from control eyes during the early healing period. The NGF content of the corneal epithelium was elevated in the wounded eye (p=0.024). Time to wound closure and rate of epithelial migration were not significantly different between the NGF treated or the NGF antibody treated, and the control BSA treated eyes. Corneal epithelial wounding increased NGF content only on the wounded side during the early healing period. Neither topical recombinant human or murine NGF affected corneal epithelial wound healing in the normal dog.

• 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.

• Korean Journal of Veterinary Research
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• v.52 no.3
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• pp.157-162
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• 2012

Corneal injury is very common clinical condition in veterinary medicine and delayed or incomplete corneal healing has the potential of vision loss due to the loss of corneal transparency. For the reconstruction of corneal epithelium, tissue graft and cell transplantation have been prosperously investigated. The purpose of this study was to evaluate the clinical value and short-term safety of application of cultured allogenic mesenchymal stem cells (MSCs) in the treatment of canine experimental corneal defect. Corneal defects were surgically generated in the central corneas of healthy beagle dogs and cultured canine allogenic MSCs were transplanted via subconjunctival injection. Although mean healing time, the rate of epithelial regeneration, and the degree of corneal transparency were not significantly improved after MSC transplantation, significant immune reaction or incompatibility reaction was not detected except transient local irritation. These results propose the possibility of MSC application as a new regenerative medicine in canine ocular disorders.

• Journal of the korean veterinary medical association
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• v.47 no.1
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• pp.42-46
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• 2011

임상에 있어 눈에 적용하는 약물의 적용은 정상적인 조직을 손상시키지 않으면서 원하는 안구 조직에 목표로 하는 용량의 안약을 전달하는 것이 목적이라고 할 수 있다. 그러나 이러한 간단한 목적도 실제로 환자의 성격, 안구 조직의 민감성, 그리고 친지질성 각막 외피세포(lipophilic corneal epithelium), 친수성 각막 그리고 공막의 간질층(hydrophilic corneal and sclera stroma), 결막의 임파 조직, 맥락막의 혈관 등, Blood-ocular barrier등 안구의 조직 자체의 장벽들로 인해 어려움을 내포하고 있는 것 역시 사실이다. 평소에 처방하고 있는 안약들을 조금 더 효과적으로 전달하기 위해 필요한 눈에 대한 이해에 대해 다루어 보도록 한다.