Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaMethods: B6LY5 mice aortic ring explants were cultured for 5 dwith the presence of 10ng/mL VEGF before TLR ligands 1-9 wereadded into culture media. Sprouting outgrowth was measured andcompared between TLR ligands treated groups and water treatedcontrol group after 5 further days of culture. Corneal angiogenesiswas induced by 3 Nylon corneal sutures on BALB/c mice beforewater, TLR3, TLR7 or TLR9 ligand was applied topically ondebrided cornea at day 0, day 2, and day 4. The corneal vesselingrowth was scored under surgical microscope after 7 days. Thevolumes of blood and lymphatic corneal vessels were quantified afterwhole mount immunofluorescence staining with CD31 and LYVE-1antibodies.Results: TLR7 and TLR9 activation suppressed vascular outgrowthfrom aortic ring explants compared to control. TLR9 engagementinhibited corneal blood vessel ingrowth after 7 d post cornealsuturing, while there was no difference between control and TLR3and TLR7 treatment groups. TLR9 activation also decreased thevolumes of blood and lymphatic corneal vessels compared to control.Conclusions: TLR9 activation could suppress aortic vessel sproutingin vitro as well as corneal hem- and lymph-angiogenesis in vivo.Commercial Relationships: Lei Liu, None; Jiahui Wu, None;Andrew D. Dick, Novartis (C), Novartis (F), GSK (F), Abbott (F)Support: Rosetrees TrustProgram Number: 2090 Poster Board Number: D0229Presentation Time: 11:00 AM - 12:45 PMSubconjunctival Cyclosporine A implants do not affect cornealneovascularisation after transplantation: results of a randomizedclinical trialClaus Cursiefen 1 , Thomas Reinhard 2 , Felix Bock 1 , Hans-UlrichProkosch 3 . 1 Dept of Ophthalmology, University of Cologne, Koln,Germany; 2 Dept. of Ophthalmology, University of Freiburg,Freiburg, Germany; 3 Medical Informatics, University of Erlangen,Erlangen, Germany.Purpose: To test whether subconjunctivally implanted CyclosporineA affects the incidence and degree of postkeratoplasty cornealneovascularisation.Methods: Prospective, randomized, controlled phase II/III clinicaltrial comprising trial sites in Germany, India and the USA. Overall,97 enrolled patients were randomized to receive either two dosages ofsubconjunctival Cyclosporine A implants (A: 36 and B: 40) orplacebo (n=21) at time of penetrating keratoplasty. The incidence anddegree of postkeratoplasty corneal neovascularisation were evaluated(LX201-01 study). A web-based image upload system wasdeveloped. Quantitative and objetive evaluation of standardizeddigital slit lamp pictures was performed using AnlysisB morphometrysofware.Results: No significant difference in the incidence and the degree ofcorneal neovascularisation was found between treatment groups andplacebo group. Mean neovascularisation was 3.2% ± 0.3 in treatmentA versus placebo (3.5% ± 0.27; p= 0.5) and 3.0% ± 0.4 and intreatment B versus placebo (3.5% ± 0.27; p= 0.3).Conclusions: Subconjunctival cyclosporine A does not affectpostkeratoplasty corneal neovascularisation.Commercial Relationships: Claus Cursiefen, Gene Signal (C),Alcon (R), Allergan (R), Bayer (R); Thomas Reinhard, None; FelixBock, None; Hans-Ulrich Prokosch, NoneClinical Trial: NCT00447187Program Number: 2091 Poster Board Number: D0230Presentation Time: 11:00 AM - 12:45 PMRole of Non-Endothelial Cells in Corneal AngiogenesisJin Zhao, Takayuki Nagasaki. Ophthalmology, Columbia University,New York, NY.Purpose: It has been suggested that growth of blood and lymphaticvessels in the cornea is assisted by non-endothelial cells in thestroma, in particular, during later stages when a vascular network isestablished by anastomosis. Supporting roles of stromal myeloid cellsand an existing nerve network have been suggested, but molecularand cellular mechanisms remain unresolved. The aim of this study isto examine evidence for the participation of stromal, non-endothelialcells during the vascular network formation.Methods: For tests of myeloid cells, four corneal angiogenesismodels in the mouse eye were used. 1) suture placement, 2) topicalapplication of ML9 (MLCK inhibitor), 3) Dstn corn1 mouse(spontaneous corneal vascularization model), 4) cornealconjunctivalization following total epithelial removal. Blood vesselgrowth was monitored in a live eye with fluorescence angiography.Cell distribution was determined by whole-mountimmunofluorescence with CD11b, CD45, CD90.2, F4/80, MHC-classII, CD31 (blood vessels) and LYVE1 (lymphatic vessels). Celldistribution was quantified in an area of vascular growth front wherea vascular network is formed by anastomosis. For a nerve guidancetest, suture was placed in the cornea of Thy1-CFP mice, and bothblood vessels (fluorescence angiography) and nerves (CFP) weretracked by time-lapse imaging in live animals.Results: Untouched, avascular corneas contained a small number ofF4/80, CD11b and MHC-class II positive cells, primarily in thelimbal area and sparsely throughout the entire cornea. CD90.2positive cell distribution had no limbal preference but was closelyassociated with sub-basal nerve plexus. After initiation ofexperimental angiogenesis, the number of cells with one or more ofmyeloid markers greatly increased within an area of vascular growth.There were frequent instances of a F4/80 positive cell being presentbetween two sprouting tips of growing vessels, appearing to bridgethem. Time-lapse imaging revealed no clear correlation betweenvessel growth and a CFP-positive nerve fiber network.Conclusions: Myeloid cells may play a supporting role in networkformation of pathological corneal vasculature either in a paracrinemanner or by means of fusion with endothelial tip cells, as reportedin developing brain. A guiding role of an existing nerve network, onthe other hand, is not supported by the data.Commercial Relationships: Jin Zhao, None; Takayuki Nagasaki,NoneSupport: Eye Surgery Fund, Research to Prevent BlindnessProgram Number: 2092 Poster Board Number: D0231Presentation Time: 11:00 AM - 12:45 PMImpaired angiogenic response in cornea by lacking TRPV1 inmiceKatsuo Tomoyose, Yuka Okada, Takayoshi Sumioka, Kumi Shirai,Shizuya Saika. Wakayama Medical University, Wakayama, Japan.Purpose: To investigate the effects of loss of Transient receptorpotential(TRP)V1 in the development of neovascularization (NV) in a cornealstroma inmice. TRP channels are polymodal receptors that are activated by ahost ofstimuli to mediate sensory transduction.Methods: Corneal NV from the limbal vessels were induced bycauterization ofthe central cornea of an eye by using disposable tool of Optemp. WTandTRPV1-null (KO) mice (n = 16 in each genotype) were used andkilled at day 3©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.