Cornea - ARVO

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ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaPurpose: To evaluate the corneal and anterior chamber condition inpatients with bullous keratopathy who underwent penetratingkeratoplaty (PK).Methods: A retrospective review of clinical records of allconsecutive patients with bullous keratopathy, referred for PK for theperiod January 2005 - December 2010, was performed. Preoperativestatus was analyzed with an emphasis on corneal condition andanterior segment abnormalities- presence and extent of cornealneovascularization, ocilar surface disorders, iris injuries withdistorted pupil, anterior synechia and subluxated intraocular lens.Results: Bullous keratophaty was present in 54 of all 141 patientswho underwent PK. Forty one of the patients had pseudophakickeratopathy, 6 pts were with aphakic and 3 with phakic keratopathy.One patient had graft failure, 1 pts- posttraumatic endothelialdecompensation and in 2 of them Fuchs endothelial decompensationwas found. Ocular surface abnormalities were diagnosed in 51 of thepatients (94%) . In 30 of the cases (56%) deep and superficial cornealneovasclarization was observed. Other pathological findingsincluded: painful epithelial bullae in 39cases (72%), subepithelial andstromal fibrosis in 18 patients (33%). In most patients all pathologicalsings were present but with different severity. Cornea was notneovascularizated and there was no fibrosis in only 5 cases. Theocular surface was not compromised in 3 of the patients. Anothermarker of advanced pathology was visual acuity: BCVA between0,05 and 0,1 in 1 patient (2%), light perception to 0,05 in 26 pts(47%) and hand movement to light perceptiont in the rest of the pts.Conclusions: The analysis of patients with bullous keratopathydemonstrated that advanced disease and severely affected corneal andanterior segment anatomy were present in all of the cases but withdifferent severity. In all patients from our studied group thepathological changes excluded the option for lamellar graftingtechniques, and PK remained the only treatment possibility. Mainreasons for the accumulation of such a pool of challenging patientswith delayed treatment are unreasonably prolonged conservativetherapy of postoperative complications and lack of donor tissue.Addressing these issues could allow the use of lamellar graftingtechniques, applicable only in the initial stages of bullouskeratophaty. At present PK remains the main therapeutic option forour patient pool.Commercial Relationships: Elena Adjievska, None; Petja I.Vassileva, None; Nikolay Surchev, None; Tatyana Hergeldzhieva-Fileva, NoneProgram Number: 2572 Poster Board Number: D0372Presentation Time: 2:45 PM - 4:30 PMHO-2 knockdown delays wound healing in Human CornealEpithelial (HCE) cells by altering the signaling of EGFR andFAK mediated pathwayAdna Halilovic, Daohong Lin, Gregory Joseph, Brian Shkolnik,Michal L. Schwartzman. Pharmacology, New York Medical College,Valhalla, NY.Purpose: Heme oxygenase (HO) represents an intrinsiccytoprotective and anti-inflammatory system. Inhibition of HOactivity significantly impairs wound healing in human cornealepithelial (HCE) cells. We have shown that HO-2 knockdown in vitroattenuates corneal wound healing and that HO-2 null mice display anaberrant corneal wound healing following injury. In this study, weinvestigated the mechanisms that may contribute to HO-2cytoprotective role in the corneal epithelium.Methods: HCE cells were stably transfected with lentivirus HO-2shRNA or non-target shRNA plasmids. Modified Boyden Chamberassay was used to study the migration of HO-2 knockdown cells.Flow cytometry cell cycle analysis was done using propidium iodidestaining. Western blot was performed to evaluate the expression ofproteins involved in migration and proliferation in HO-2 shRNA andnon-target shRNA cells.Results: HO-2 knockdown, using virus-mediated shRNA, impairedHCE wound healing by 35% and 25% at 24h and 48h after injury,respectively. Furthermore, HO-2 knockdown inhibited basal andEGF(10ng/mL)-stimulated HCE migration by 50% and 40%,respectively and was associated with a 35% decrease in p-Akt and45% decrease in p-EGFR levels. Flow cytometric analysis showedthat EGF-stimulated cell cycle progression was attenuated in HO-2knockdown cells. Immunofluorescence of HO-2shRNA and controltransfected cells revealed a cytoplasmic distribution of p-FAK. Uponinjury, control cells displayed typical p-FAK as focal adhesion pointsin the membrane, whereas in HO-2 knockdown cells p-FAK had nofocal distribution.Conclusions: Knockdown of HO-2 results in attenuated cornealepithelial wound healing. Deficiency of HO-2 impairs wound healingby a mechanism that involves alterations in ERK, Akt and FAKsignaling pathways, all of which have been shown to play animportant role in growth factor-mediated cell migration andproliferation following injury.Commercial Relationships: Adna Halilovic, None; Daohong Lin,None; Gregory Joseph, None; Brian Shkolnik, None; Michal L.Schwartzman, NoneSupport: NIH Grant EY06513 (MLS)Program Number: 2573 Poster Board Number: D0373Presentation Time: 2:45 PM - 4:30 PMDecreased Incidence of Perioperative Corneal Injuries Followingan Ophthalmology Educational Initiative For AnesthesiaProviders at an Academic Medical CenterAmanda L. Ely, Ingrid U. Scott, Tabassum F. Ali, Denise Kerchner,David Liang, Michael Wilkinson. Ophthalmology, Penn StateHershey Ophthalmology, Hershey, PA.Purpose: To investigate whether an ophthalmology educationalinitiative directed toward anesthesia providers can decrease theincidence of perioperative corneal injury at an academic medicalcenter.Methods: Prospective chart review of consults placed forperioperative eye pain resulting from corneal injury between July 1,2011 and January 4, 2012 (including patient and surgical risk factorsfor corneal injury as reported in the literature). An ophthalmologyeducational initiative, including a Grand Rounds lecture anddistribution of educational materials regarding perioperative ocularcare, was provided to anesthesia providers on January 5, 2012 basedon the Mayo Clinic Model (Martin et al., Anesthesiology 2009).Beginning on January 5, 2012, an email was sent to the attending andresident physician of any patient who experienced a perioperativecorneal injury; the email notified the physicians of the corneal injuryand provided information to the physicians on how such injuries maybe prevented in the future. Prospective chart review of consults(identical to the pre-initiative data collection) was then performedbetween January 6, 2012 through June 30, 2012. The rates ofperioperative corneal injury pre- versus post-initiative werecompared.Results: The rate of perioperative corneal injury was 0.35% beforethe educational initiative compared with 0.18% following theinitiative (p=0.04). The proportions of corneal injuries that wereassociated with risk factors reported in the literature for such injurieswere not significantly different pre- versus post-initiative except forlength of surgical case. Pre-initiative a higher proportion of cornealinjuries occurred after a surgery
ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaConclusions: An ophthalmology educational initiative directedtoward anesthesia providers was associated with a significantdecrease in the incidence of perioperative corneal injury. Thisinformation may be helpful for other medical centers in order toemphasize the potential impact of educational initiatives to preventperioperative corneal injury.Commercial Relationships: Amanda L. Ely, None; Ingrid U.Scott, None; Tabassum F. Ali, None; Denise Kerchner, None;David Liang, None; Michael Wilkinson, NoneSupport: n/aProgram Number: 2574 Poster Board Number: D0374Presentation Time: 2:45 PM - 4:30 PMMOLECULAR IDENTIFICATION AND FUNCTIONALCHARACTERIZATION OF THE VITAMIN CTRANSPORTERS EXPRESSED BY HUMAN CORNEALEPITHELIAL CELLS (HCEC)Varun Khurana, Aswani Dutt Vadlapudi, Ramya Krishna Vadlapatla,Dhananjay Pal, Ashim K. Mitra. Pharmaceutical Sciences, Universityof Missouri Kansas City, Kansas City, MO.Purpose: The objective of this research was to functionallycharacterize the Sodium Dependent Vitamin C Transporter (SVCT)in human corneal epithelial cells (HCEC). This nutrient transportercan be targeted to improve the delivery of drugs that get effluxed outby the efflux transporters (P-gp, MRP and BCRP).Methods: Uptake was performed with human corneal epithelial cells(HCEC). Uptake of [14C] ascorbic acid ([14C] AA) was studied inthe absence and presence of excess unlabelled AA, anion transporterinhibitors. Effect of pH, sodium and temperature on the uptake of[14C] AA was also studied. Concentration dependency studies(0.1µM-1000µM) were performed. Role of cellular protein kinasemediated pathways and Ca2+/CaM mediated pathways on theregulation of AA uptake has also been studied. Reverse transcriptionpolymerase chain reaction (RT-PCR) for SVCT was carried out onRNA isolated from HCEC.Results: Uptake of [14C] AA was found to be saturable with a K¬mof 144.87 µM and Vmax of 38.33 pmol/mg protein/min. Uptake of[14C] AA was pH, sodium, energy and temperature dependent. Themembrane transporter was also under the regulation of cellularprotein kinase C (PKC), protein kinase A (PKA), Ca2+/CaMmediated pathways and metabolic inhibitors. Uptake of [14C] AAwas significantly inhibited in the presence of different concentrationsof cold L- ascorbic acid and D-Iso ascorbic acid. The observedpermeability values for transepithelial transport were 2.29±0.56×10-4, 2.07±0.45×10-4, 1.49±0.38×10-4, 1.8±0.24×10-4 and1.57±0.66×10-4 cm/s for [14C] AA, L-AA (100µM), L-AA(500µM),D-Iso AA(100µM) and D-Iso AA (500µM) respectively. Ethidiumbromide staining of the gel showed a major 626-bp bandcorresponding with the amplified SVCT2 cDNA.Conclusions: Sodium Dependent Vitamin C Transporter (SVCT)which plays an important role in the uptake of ascorbic acid has beenidentified and functionally characterized in HCE cell line. Thisnutrient transporter can be targeted to improve the delivery of drugsthat acts as substrate for the efflux transporters (P-gp, MRP andBCRP). The study also provides useful information on the substratespecificity of this carrier system. This study suggests that HCE cellsexpress the hSVCT2 system and may serve as a useful in vitroexperimental model to study the permeability of ascorbic acidconjugated prodrugs.Commercial Relationships: Varun Khurana, None; Aswani DuttVadlapudi, None; Ramya Krishna Vadlapatla, None; DhananjayPal, None; Ashim K. Mitra, NoneSupport: NIH R01EY09171-16 and NIH R01EY010659-14Program Number: 2575 Poster Board Number: D0375Presentation Time: 2:45 PM - 4:30 PMAn animal model for epithelial downgrowthJessica E. Weinstein, Matthew J. Weiss, Jeffrey L. Goldberg.Ophthalmology, Bascom Palmer Eye Inst, Univ of Miami, Miami,FL.Purpose: Epithelial downgrowth is a rare but devastatingcomplication of penetrating trauma or surgery. It can result in visionloss, intractable glaucoma or corneal decompensation. Treatmentmethods are usually non-curative and are often followed byrecurrence. An animal model of epithelial downgrowth is essentialfor the development of future diagnostics and therapies, but to datethere are few options. Here we developed a rat model of epithelialdowngrowth.Methods: Mouse corneal epithelial cells (MCECs) were injected intothe anterior chamber of Sprague-Dawley rats and observed over 4weeks. Anterior chamber ocular coherence tomography (OCT) andintra-ocular pressure (IOP) measurements were done at baseline, 2and 4 weeks. In each rat, the contralateral eye was used as a control.OCT measurements of corneal thickness were analyzed usingMatLab software. Rat eyes were fixed by immersion for 2-3 hourswith 4% PFA and then embedded in OCT Tissue-Tek and frozen.Whole eyes were sectioned. Half the representative samples werestained for H&E and the rest were stained withimmunohistochemistry with corneal epithelial cell marker antibodies.Results: H&E staining showed presence of membranes andadditional cells in the anterior chamber. DAPI-stained nuclei wereevident in the membranes in the experimental eyes when compared tocontrol eyes. Anterior chamber OCT showed thicker corneas inexperimental eyes compared to control eyes. IOPs were notsignificantly elevated in experimental eyes compared to control eyes.Conclusions: A novel rat model was developed for epithelialdowngrowth which mimicked some features of the typical humandisease. Further experiments with longer time-points are planned toinvestigate whether IOP increases eventually. Development of thisepithelial downgrowth model may lead to new opportunities fordiagnostic and treatment development.Commercial Relationships: Jessica E. Weinstein, None; MatthewJ. Weiss, ASCRS Foundation Grant (F); Jeffrey L. Goldberg, NoneSupport: RPB Medical Student Fellowship; ASCRS GrantProgram Number: 2576 Poster Board Number: D0376Presentation Time: 2:45 PM - 4:30 PMDescriptive Study of Ocular Surface changes in Patients withCongenital GlaucomaSimone Finzi, Ruth M. Santo, Monique Matsuda, Ernst W. Oltrogge,Marcio H. Mendes, Fernando E. Naves, Rodrigo P. Azevedo,Frederico Lazar, Bruno C. Cardoso, Alberto Betinjane.Ophthalmology, University of Sao Paulo, Sao Paulo, Brazil.Purpose: Recent studies show a relationship between POAG andlacrimal dysfunction. Several factors may lead to alteration of theocular surface (OS) in patients with primary congenital glaucoma(PCG). The purpouse of this study is to elaborate a clinicaldescription of the ocular surface (OS) in patients with GCPMethods: After informed consent, 34 patients diagnosed with PCGwere included in the study. Ophthalmologic examination wasperformed, and clinical evaluation of OS. The patients weresubmitted to a questionnaire to evaluate symptoms of dry eyeResults: We examined 34 patients (60 eyes) diagnosed with PCG to17.97 ± 7.91 years. 17 patients without use of eyedrop for at least 1year and 17 patients using eye drops, these 9 patients (64%) using 3or more drops. The patients had chronic use of eye drops with a mean©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.
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