Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - Corneain HaCaT cells.The excipients did not interfere with effects of BMP-7 and also didnot affect cell viability.Conclusions: BMP-7 with excipients has the ability to reducefibrosis and also the excipients contributed to the stabilization ofBMP-7. As such, it seems possible that this technology can beapplied to eye drops and become a new innovative treatment inpharmaceutical markets.Commercial Relationships: Jin-Wook Jang, None; Hansoo Kim,None; Chan-Young Cho, None; Jinkuk Kim, None; Ju-WoongJang, None; Young-Sik Kim, None; Ynag-je Cho, NoneSupport: 10037842 (Ministry of Knowledge Economy, Repulic ofKorea)Program Number: 3881 Poster Board Number: D0125Presentation Time: 2:45 PM - 4:30 PMGalectin-3 Enhanced Epithelialization in Explanted MonkeyCorneas with Alkali BurnAtsuko Fujii 1, 2 , Thomas R. Shearer 2 , Mitsuyoshi Azuma 1, 2 .1 Laboratory of Ocular Sciences, Senju Pharmaceutical Co, Ltd.,Beaverton, OR; 2 Department of Integrative Biosciences, OregonHealth & Science University, Portland, OR.Purpose: Following an injury leading to the loss of the cornealepithelium, the remaining epithelial cells immediately attempt toclose the defect. Poor healing of epithelial wounds is a major clinicalproblem, leading to persistent epithelial defects and ulceration. Wepreviously showed that carbohydrate-binding protein galectin-3 (Gal-3) enhanced wound closure in explanted monkey corneas laceratedby n-heptanol. Alkali injuries of the eye often cause extensivedamage to the cornea due to rapid penetration and damage to deeperocular structures. The purpose of the present experiment was to studyGal-3 in the more severe alkali burn model, and to compare results tothe n-heptanol model.Methods: An alkali burn was created by a 60 sec application of a 7.5mm diameter filter disc soaked in 1N sodium hydroxide onto thecentral cornea of enucleated monkey eyes. The corneas were thenexcised, incubated for various times with or without recombinantGal-3, and stained with 1% sodium fluorescein. Corneal woundclosure was quantified by digital image analysis.Results: After an alkali burn, the corneal wound area became smallerin a time-dependent manner. Exogenous recombinant Gal-3 enhancedthis wound closure to the same extent as in corneas wounded with n-heptanol. However, data suggested that the rates of healing weredifferent in the two models.Conclusions: Exogenous Gal-3 showed a beneficial effect on closureof wounds caused by either alkali or n-heptanol. In both cases, thismay be because Gal-3 binds to ECMs such as laminin and collagenand promotes lamellipodia formation by cross-linking to α3 integrin.Although initial cytokines released from local cells may be differentin our two chemical models, the results indicate that Gal-3 may be acandidate drug to enhance epithelialization in human cornea damagedby variable causes.Dr. Shearer receives a research contract and consulting fees from, andDr. Azuma and Ms. Fujii are employees of, Senju Pharmaceutical Co.Ltd.Commercial Relationships: Atsuko Fujii, Senju PharmaceuticalCo., Ltd. (E); Thomas R. Shearer, Senju Pharmaceutical Co., Ltd.(F), Senju Pharmaceutical Co., Ltd. (C); Mitsuyoshi Azuma, SenjuPharmaceutical Co., Ltd. (E)Program Number: 3882 Poster Board Number: D0126Presentation Time: 2:45 PM - 4:30 PMMicroRNA-182 Inhibits Human Corneal Epithelial CellProliferation and MigrationDongsheng Yan 1, 2 , Xiaoyan Chen 1, 2 , Jiao Wang 1, 2 , Lili Tu 1, 2 .1 School of Optometry and Ophthalmology, Wenzhou MedicalCollege, Wenzhou, China; 2 State Key Laboratory Cultivation Baseand Key Laboratory of Vision Science, Ministry of Health of P. R.China, Zhejiang Provincial Key Laboratory of Ophthalmology andOptometry, Wenzhou, China.Purpose: MicroRNAs (miRNAs) are endogenous short (~22)nucleotide RNAs which inhibit protein translation through binding totarget mRNAs. Recent studies have demonstrated that miR-182 canregulate tumor cell proliferation and migration. The role of miR-182in corneal wound healing, however, remains unclear. In the presentstudy, we investigated the function of miR-182 in human cornealepithelial cells.Methods: Realtime RT-PCR was performed to detect the expressionof miR-182 in mouse corneal epithelium during wound healingprocess. Human corneal epithelial cells were transfected with miR-34a. MTS and wound-healing assay was carried out to evaluate theeffect of miR-182 on human corneal epithelial cell proliferation andmigration, respectively. The expression of c-Met protein wasdetermined by Western blotting.Results: miR-182 was downregulated during corneal wound healingprocess. Transfection of miR-182 into human corneal epithelial cellsled to a significant decrease in cell proliferation and migration. miR-182 downregulated the expression of c-Met by Western blot analysis.Conclusions: Our results demonstrated that miR-182 inhibitedhuman corneal epithelial cell proliferation and migration bydownregulation of c-Met. This indicates that miR-182 may play animportant role in corneal wound healing process.Commercial Relationships: Dongsheng Yan, None; XiaoyanChen, None; Jiao Wang, None; Lili Tu, NoneSupport: National Natural Science Foundation of China(81071682&81272286)Program Number: 3883 Poster Board Number: D0127Presentation Time: 2:45 PM - 4:30 PMThe effects of IL-6 receptor blockade on gene expressions inexperimental corneal alkali burnSatoshi Sugaya, Tohru Sakimoto, Ai Yamada, Takako Ohnishi, AkikoIshimori, Mitsuru Sawa. Nihon University school of medicine,Tokyo, Japan.Purpose: Using experimental corneal alkali burn model, wepreviously reported the suppressive effect of topical instillation ofanti-Interleukin-6 receptor (IL-6R) antibody on infiltrations ofinflammatory cells. Using this model, we investigated the geneexpressions of inflammation-related factors and chemotactic factors.Methods: Unilateral eye of corneal alkali burn was made using afilter paper dipped in 1 N NaOH solution using BALB/c mice. Threeeyes of 3 mice were treated by topical instillation of anti-IL-6Rantibody solution (MR16-1, 2 µg/µL) from day 5 to day 28 afterwounding. (MR16-1 group)(from day 5 to day 14; 3 times/day, fromday 15 to day 28; 2 times/day) The control group (3 eyes of 3 mice,PBS group) underwent topical 0.01M PBS (pH 7.4) solution. Themice were euthanatized on day 28 and 2-μm-thick sections weremade. Corneal stroma section was made using laser capturemicrodissection system, and total RNA in the specimens wasdetermined by quantitative PCR array method (Mouse Th17Response Array, QIAGEN).Results: In MR16-1 group, MMP (matrix metalloproteinase)-13(6.39±3.54; mean ± SD) expression was decreased by six timescompared with that in PBS group. The chemotactic factors ofmacrophage, MCP (monocyte chemotactic protein)-1 (5.83±1.95)©2013, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permissionto reproduce any abstract, contact the ARVO Office at arvo@arvo.org.