• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.803-808
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• 2010

Poly-${\gamma}$-glutamic acid (${\gamma}$-PGA) is a natural edible polypeptide in which glutamate is polymerized via ${\gamma}$-amide linkages. First, we assessed the eye irritancy potential of ${\gamma}$-PGA in rabbits. Additionally, we studied the effects of ${\gamma}$-PGA on corneal wound healing, due to the anti-inflammatory properties and water retaining abilities of ${\gamma}$-PGA. In this study, the effects of ${\gamma}$-PGA on corneal wound healing after an alkali burn were evaluated. Thirty eyes wounded by alkali burning in 30 white rabbits were divided into three groups: group A was treated with 0.1% 5,000 kDa ${\gamma}$-PGA for 2 days; group B was treated with 0.1% hyaluronic acid; and group C was not treated, as a control. The area of corneal epithelial defect was examined at 12, 24, 30, 36, 42, and 48 h after corneal alkali wounding to determine initial wound healing. We found that ${\gamma}$-PGA promoted corneal wound healing, compared with controls, and showed similar effects to hyaluronic acid. These results indicate that ${\gamma}$-PGA stimulates corneal wound healing by an anti-inflammatory effect and enhancing cell migration and cell proliferation. ${\gamma}$-PGA is a promising biomaterial that may be a substitute for hyaluronic acid in corneal wound healing treatment.

• 한국임상수의학회지
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• 제18권4호
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• pp.418-423
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• 2001

각막상피의 창상치유에 중요한 역할을 하는 것으로 알려진 Nerve Growth Factor(NGF)의 내인성 변화와 국소적 투여효과를 알아보았다. 6마리의 Beagle 을 사용하여 비교군인 오른쪽 눈에만 6mm의 기계적인 각막 창상을 만들었고 왼쪽 눈은 대조군으로 사용하였다. 눈물은 수술후 1주일동안 매일, 각막 상피는 수술중 그리고 수술후 7일에 각각 채취하여 NGF 농도를 enzyme-linked immunosorbent assay (ELISA)를 사용하여 측정하였다. 2차 실험으로, 16마리의 Bealgle을 사용하여 오른쪽 눈에 위와 동일한 창상을 만든 후, 창상 부위가 회복될 때까지 recombinant human (rh) NGF (n=4), murine NGF (n=6) 그리고 anti-NGF blocking antibody (n=6)를 6시간마다 점안하였으며, 왼쪽 눈은 bovine serum albumin (BSA)를 점안하여 대조군으로 사용하였다. 창상 면적은 NIH image software를 이용하여 분석하였다. 대조군에 비하여, 비교군의 눈물내 NGF는 치유초기에 현저하게 증가하였으며, 각막 상피의 NGF도 유의적인 증가를 보였다. 그러나 rh NGF, murine NGF, anti-NGF blocking antibody처치군과 BSA처치군 간에는 각막창상 치유에 유의적인 차이가 인정되지 않았다. 각막창상후, rh NGF 혹은 murine NGF의 추가적인 국소점안은 정상개의 각막상피 치유에 효과가 없었으나, 창상후 초기 치유기동안 눈물과 손상받은 각막상피에서 내인성 NGF의 급격한 보상성 증가로 미루어 NGF는 각막 상피의 창상치유에 중요한 역할을 하는 것으로 사료된다.

• 한국광학회지
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• 제34권2호
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• pp.45-52
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• 2023

본 연구에서는 다양한 파장의 저출력 light-emitting diode (LED)를 이용한 photobiomodulation (PBM)이 각막 상처 치유에 미치는 영향을 분석하였다. 각막상피세포에 623 nm에서 940 nm 범위의 파장의 LED를 조사한 결과, 유의미한 세포독성 영향을 미치지 않는 것을 확인하였다. PBM의 세포이동 촉진 효과를 세포 이동능 평가 시험을 통해 분석한 결과 623 nm 파장의 광조사에 의한 PBM이 세포이동을 크게 증가시키고 상처 치유를 촉진하는 것으로 나타났다. 또한, 세포이동 및 상처 치유와 관련된 유전자의 발현을 분석한 결과, 623 nm 파장의 광조사에 의한 PBM이 세포 증식과 세포 외 기질 분해를 촉진하는 것으로 알려진 FGF-1과 MMP2 유전자의 발현을 상향 조절한다는 사실을 발견했다. 이러한 연구 결과는 특정 파장, 특히 623 nm 파장의 저출력 빛을 이용한 PBM이 각막 손상 치료에 활용될 수 있는 가능성을 시사한다.

• BMB Reports
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• 제42권12호
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• pp.800-805
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• 2009

This study investigated the effect of subconjunctival injections of bevacizumab, an anti-VEGF antibody, on processes involved in corneal wound healing after alkali burn injury. Mice were divided into three groups: Group 1 was the saline-treated control, group 2 received subconjunctival injection of bevacizumab 1hr after injury and group 3 received bevacizumab 1 hr and 4 days after injury. Cornea neovascularization and opacity were observed using a slit lamp microscope. Corneal repair was assessed through histological analysis and immunostaining for CD31, $\alpha$-SMA, collagen I, and TGF-$\beta$2 7 days post-injury. In group 3, injection of bevacizumab significantly lowered neovascularization and improved corneal transparency. Immunostaining analysis demonstrated a reduction in CD31, $\alpha$-SMA and TGF-$\beta$2 levels in stroma compared to group 1. These results indicate that bevacizumab may be useful in reducing neovascularization and improving corneal transparency following corneal alkali burn injury by accelerating regeneration of the basement membrane.

• Medical Lasers
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• 제10권1호
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• pp.22-30
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• 2021

Background and Objectives Ocular alkali burns cause severe damage to the ocular tissues and vision loss. Solcoseryl is a standardized calf blood extract that normalizes the metabolic disturbance and aids in maintaining the chemical and hormonal balance and has been used to treat burns in various tissues. This study examined the effects of Solcoseryl on a rat corneal alkali burn model. Materials and Methods Twenty rats were assigned randomly to four equal groups, including alkali burn, hyaluronic acid, Solcoseryl eyedrop, and Solcoseryl gel. A corneal alkali burn was induced by a NaOH-soaked paper disc. The treatments were given twice a day, every day. The wound area was measured after 24 and 48 hours, and the degree of neovascularization and corneal opacity were scored every week. The rats were sacrificed after three weeks for immunohistochemistry (IHC) to compare the level of inflammatory cytokines, IL-1β, IL-6, and TNF-α. The thickness of the retinal layers was compared to observe any changes in the retina. Results The use of Solcoseryl on corneal alkali burn accelerated wound healing with less neovascularization, greater opacity, and less cataract. IHC showed that the inflammation of the cornea was controlled by both the hyaluronic acid and Solcoseryl treatments. On the other hand, the inflammation had spread to the retina. When the dosage forms were compared, eyedrops were more effective on corneal inflammation, while the gel-type had a greater effect on retinal inflammation. Conclusion Solcoseryl was effective in accelerating the wound healing rate on a corneal alkali burn but could not prevent the spread of inflammation from the cornea to the retina. Eyedrops were more effective on inflammation in the cornea, and the gel was more effective in the retina.

• 한국임상수의학회지
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• 제37권4호
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• pp.198-203
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• 2020

Synovial fluid (SF) contains various factors which may be helpful for corneal stromal healing, such as cytokines, growth factors, hyaluronic acid, and proteins. Therefore, the purpose of this study was to determine the effect of SF on proliferation and migration in canine keratocytes. In order to evaluate the degree of proliferation and migration, canine keratocytes were cultured in DMEM containing 1%, 3%, 5%, or 10% SF. Real-time PCR was performed in a control group and the group treated with 5% SF, in order to measure the expression levels of factors associated with corneal wound healing. These factors included interleukin-1α (IL-1α), hepatocyte growth factor (HGF), transforming growth factor-β (TGF-β), and α-smooth muscle actin (SMA-α). Proliferation assays demonstrated that proliferation was significantly enhanced in groups treated with greater than 3% SF, as compared with that of the control group. In addition, migration in all SF-treated groups was significantly increased as compared with migration in the control group, as measured by migration assays. mRNA expression of IL-1α and HGF was significantly increased and mRNA expression of TGF-β and SMA-α was significantly decreased in the cells treated with 5% SF. These findings suggest that SF may promote corneal wound healing.

• BMB Reports
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• 제46권4호
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• pp.195-200
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• 2013

To understand the corneal regeneration induced by bevacizumab, we investigated the structure changes of stroma and basement membrane regeneration. A Stick soaked in 0.5 N NaOH onto the mouse cornea and 2.5 mg/ml of bevacizumab was delivered into an alkali-burned cornea (2 ${\mu}l$) by subconjunctival injections at 1 hour and 4 days after injury. At 7 days after injury, basement membrane regeneration was observed by transmission electron microscope. Uneven and thin epithelial basement membrane, light density of hemidesmosomes, and edematous collagen fibril bundles are shown in the alkali-burned cornea. Injured epithelial basement membrane and hemidesmosomes and edematous collagen fibril bundles resulting from alkali-burned mouse cornea was repaired by bevacizumab treatment. This study demonstrates that bevacizumab can play an important role in wound healing in the cornea by accelerating the reestablishment of basement membrane integrity that leads to barriers for scar formation.

• The Korean Journal of Physiology and Pharmacology
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• 제21권2호
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• pp.189-195
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• 2017

This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.

• 대한수의학회지
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• 제63권2호
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• pp.20.1-20.4
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• 2023

A 10-year-old, spayed female, Maltese dog presented with a 2-day history of severe left eye squint. Slit-lamp biomicroscopy showed a deep corneal defect stained into a doughnut shape together with hypopyon in the anterior chamber. Based on these results, a diagnosis of descemetocele and uveitis in the left eye was made. Deep keratotomy combined with a nictitating membrane flap effectively resolved the descemetocele without complications. The surgical procedures performed on this patient were easy and effective, and could be used as an alternative to graft surgery for descemetocele treatment.

• Ocean and Polar Research
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• 제30권4호
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• pp.467-472
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• 2008

Antarctic krill has a strong proteolytic enzyme system, which comes from a combination of several proteases. This powerful activity can be easily detected by krill's superior post mortem autolysis. Mammalian skin consists of epidermis and dermal connective tissue, and functions as a barrier against threatening environments. A clot in a wound site of the skin should be removed for successful skin regeneration. Epithelial cells secrete proteases to dissolve the clot. In previous studies Antarctic krill proteases were purified and characterized. The proteolytic enzymes from Antarctic krill showed higher activity than mammalian enzymes. It has been suggested that these krill clean up the necrotic skin wound to induce a natural healing ability. The enzymes exhibited additional possibilities for several other biomedical applications, including dental plaque controlling agent and healing agent for corneal alkali burn. Considering that these versatile activities come from a mixture of several enzymes, discovering other proteolytic enzymes could be another feasible way to enhance the activity if they can be used together with krill enzymes. Molecular cloning of the krill proteases should be carried out to study and develop the applications. This review introduces possible roles of the unique Antarctic krill proteases, with basic information and suggestion for the development of an application to skin regeneration.