Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - CorneaCommercial Relationships: Ricardo Nose, None; Fabio B. Daga,None; Adriana S. Forseto, None; Gustavo Victor, None; Sidney J.Sousa, None; Walton Nose, None; Niro Kasahara, NoneProgram Number: 3892 Poster Board Number: D0136Presentation Time: 2:45 PM - 4:30 PMEffect of human albumin in combination of blood derivatives richin growth factors in the wound healing capability of cornealepithelial cellsNoelia Andollo 1 , Vanesa Freire 1, 4 , Arturo E. Grau 3 , JaimeEtxebarria 1, 5 , Juan A. Duran 2, 4 , Maria-Celia Morales 5 . 1 CellBiology And Histology, University of The Basque Country,BioCruces Health Research Institute, Leioa, Spain; 2 Ophthalmology,University of the Basque Country, BioCruces Health ResearchInstitute, Leioa, Spain; 3 Hospital Sótero del Rio, Santiago, Chile; 4 R& D Dept., Instituto Clínico-Quirúrgico de Oftalmología, Bilbao,Spain; 5 University Hospital of Cruces, BioCruces Health ResearchInstitute, Barakaldo, Spain.Purpose: Blood derivatives rich in growth factors are goodpromoters of corneal wound healing. Our aim is to test in vitro thecapability of human albumin as a synergic compound combined witha blood derivative rich in growth factors to improve the healing rateof corneal epithelial cells.Methods: In vitro proliferation and wound healing experiments wereperformed using the human corneal epithelial HCE cell line. Westudied the following treatments: 1) 20% Serum of Plasma Rich inGrowth Factors (s-PRGF) 2) 50% s-PRGF 3) 10% Human serumalbumin (albumin) 4) 20% s-PRGF+10% albumin 5) 50% s-PRGF+10% albumin 6) 10% FBS and 7) 1% bovine serum albumin(BSA) as control. To manufacture s-PRGF whole blood was collectedby venipuncture from healthy volunteers.Results: The results show that human albumin induces highlysignificant higher proliferation of HCE cells than any dose of s-PRGFafter 72 hours of treatment. Moreover, no significant differences arefound among human albumin, 20% s-PRGF+albumin and 50% s-PRGF+albumin. Conversely, 20% and 50% s-PRGF treatmentsimprove cell migration with respect to the rest of the treatments, withstatistically significant differences. Treatments of albumin, 20% s-PRGF+albumin and 50% s-PRGF+albumin do not show significantdifferences among them.Conclusions: Human albumin by itself induces higher proliferationof epithelial corneal cells but lower cell migration, compared with theeffect of the blood derivative s-PRGF. In addition, combination ofboth does not improve proliferation and affects negatively migration.Commercial Relationships: Noelia Andollo, None; Vanesa Freire,None; Arturo E. Grau, None; Jaime Etxebarria, bioftalmik (C);Juan A. Duran, None; Maria-Celia Morales, NoneProgram Number: 3893 Poster Board Number: D0137Presentation Time: 2:45 PM - 4:30 PMCryopreservation Preserves the Structural Integrity, BiochemicalComponents and Biologic Function of Amniotic MembraneTissueEk Kia Tan 1, 2 , Marissa T. Cooke 3 , Christian Mandrycky 3 , Hua He 1, 2 ,Julie O'Connell 4 , Todd C. McDevitt 3, 5 , Scheffer C. Tseng 1, 2 . 1 OcularSurface Research and Education Foundation, Miami, FL; 2 Researchand Development, TissueTech, Inc., Miami, FL; 3 Wallace H. CoulterDepartment of Biomedical Engineering, Georgia Institute ofTechnology and Emory University, Atlanta, GA; 4 Amniox Medical,Marietta, GA; 5 Parker H. Petit Institute for Bioengineering andBioscience, Georgia Institute of Technology, Atlanta, GA.Purpose: The therapeutic potential of amniotic membrane (AM) hasbeen examined in a variety of clinical indications, particularly inophthalmology where AM has been used extensively for indicationssuch as pterygium, conjuntivochalasis, and corneal defects. To date,there has been no comprehensive study comparing cryopreservedamniotic membrane processed with the CryoTek method and freshAM.Methods: Fresh AM, either thin or thick, and fresh amniochorionwere compared to CryoTek processed thin and thick AM andamniochorion. Histological properties were assessed by hematoxylinand eosin, Masson’s Trichrome, and Safranin O staining.Biochemical properties were measured by contents of protein,albumin, and hyaluronan (HA) with their molecular weight spectrum.Functional assays compared macrophage viability and proliferation,and human corneal fibroblast TGF-β1 induction.Results: Histochemical staining confirmed that the cryopreservationprocess did not alter the tissue architecture or collagen andglycosaminoglycan content. Biochemically, cryopreservation reducedtotal protein and human serum albumin contents, but retained highmolecular weight hyaluronan species including HC-HA complex,known to exert anti-inflammatory and anti-scarring effects. Bothfresh and cryopreserved AM water-soluble extracts similarlysuppressed viability and proliferation of RAW264.7 macrophages,and dose-dependently inhibited the TGF-β1 promoter activity inhuman corneal fibroblasts.Conclusions: These results collectively indicate thatcryopreservation by the CryoTek method effectively preserveshistological, biochemical and functional components of the AMtissue.Commercial Relationships: Ek Kia Tan, TissueTech, Inc. (E);Marissa T. Cooke, None; Christian Mandrycky, None; Hua He,TissueTech, Inc. (E); Julie O'Connell, None; Todd C. McDevitt,Amniox Medical (C); Scheffer C. Tseng, NIH, NEI (F), TissueTech,Inc. (F), TissueTech, Inc. (E), TissueTech, Inc. (P)Support: Research Alliance Venture Lab Grant #398, Atlanta, GAand a research grant from TissueTech, Inc., Miami, FL.Program Number: 3894 Poster Board Number: D0138Presentation Time: 2:45 PM - 4:30 PMBiological Differences between Cryopreserved and DehydratedAmniotic Membrane Tissue GraftsLorraine S. Chua 1, 2 , Marissa T. Cooke 3 , Christian Mandrycky 3 , EkKia Tan 1, 2 , Julie O'Connell 4 , Scheffer C. Tseng 1, 2 , Todd C.McDevitt 3, 5 . 1 Ocular Surface Research & Education Foundation,Miami, FL; 2 Research & Development, TissueTech, Inc., Miami, FL;3 Wallace H. Coulter Department of Biomedical Engineering, GeorgiaInstitute of Technology and Emory University, Atlanta, GA;4 Amniox Medical, Marietta, GA; 5 Parker H. Petit Institute forBioengineering and Bioscience, Georgia Institute of Technology,Atlanta, GA.Purpose: Amniotic membrane (AM) processing methods candramatically impair both the structural integrity, and biologicalactivity, of critical matrix cell signaling factors essential for theintended use of the product. To analyze the effect of different AMavailable commercially for the ophthalmology market on conservingthe therapeutic potential of the tissue, we compared cryopreserved(CryoTek) and dehydrated (Purion®) processed tissue grafts inphysical and biochemical assays.Methods: Cryopreserved thin (CT-Thin) and thick (CT-Thick)tissues were compared to dehydrated, (EF) and (AF) AM tissuegrafts. Structural properties of cryopreserved and dehydrated AMwere assessed by histological staining while biochemical propertieswere measured in soluble tissue extracts by comparing hyaluronan(HA) content and molecular weight (MW) spectrum; and criticalproteoglycan (HC-HA) and protein (PTX3) signaling factors.©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.