733 TISSUE ENGINEERING OF AN ORAL MUCOSA EQUIVALENT BY EVALUATION OF DIFFERENT BIOMATRICES

Abstract

You have accessJournal of UrologyPediatrics: Basic Research1 Apr 2012733 TISSUE ENGINEERING OF AN ORAL MUCOSA EQUIVALENT BY EVALUATION OF DIFFERENT BIOMATRICES Daniela Flesch, Elena Frerick-Ochs, Celine Pfeifer, Ronald E. Unger, Annette Schröder, Raimund Stein, Joachim W. Thüroff, and Walburgis Brenner Daniela FleschDaniela Flesch Mainz, Germany More articles by this author , Elena Frerick-OchsElena Frerick-Ochs Mainz, Germany More articles by this author , Celine PfeiferCeline Pfeifer Mainz, Germany More articles by this author , Ronald E. UngerRonald E. Unger Mainz, Germany More articles by this author , Annette SchröderAnnette Schröder Mainz, Germany More articles by this author , Raimund SteinRaimund Stein Mainz, Germany More articles by this author , Joachim W. ThüroffJoachim W. Thüroff Mainz, Germany More articles by this author , and Walburgis BrennerWalburgis Brenner Mainz, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.818AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Oral mucosa has been used for urethral reconstruction in case of hypospadia for more than a decade. Harvesting large specimens of oral mucosa when reconstructing large sections of the urethra is technically challenging. Besides, it causes great discomfort, pain and secondary scarring to the patient. Therefore alternative techniques are required. One of these techniques is the construction of a tissue engineered oral mucosa, composed of a biomatrix seeded with primary cells. METHODS From human oral mucosa specimens, fibroblasts and epithelial cells were isolated and cultured separately. These primary cells were seeded on two collagen matrices (BioGide, BioGidePro) and on SIS (small intestinal mucosa). For coculture, fibroblasts were seeded on one side of the matrix and epithelial cells on the other side. After 21 days of cultivation the seeded matrices were analysed by fluorescent-vitality dyeing and immunohistochemically staining of Cytokeratin 13, Collagen IV and Tenascin. To achieve prevascularisation of the oral mucosa equivalent microvascular endothelial cells (HDMECs) were seeded together with fibroblasts on the porous side of the matrix. To analyse the triculture system a PECAM staining was performed and examined via confocal laser scanning microscope. Additionally the triculture was stained immunohistochemical. RESULTS Cocultures of the three matrices (BG, BGP, and SIS) showed typical characteristics of native oral mucosa. Staining with Cytokeratin-13 confirmed the formation of a multilayer epithelium on the cell occlusive site of the matrix. Between epithelium and matrix a basal membrane could be detected by immunohistochemical staining of collagen IV. Additionally fibroblast infiltration into the matrix could be observed. Fibroblasts produced new extracellular matrix, detected by Tenascin staining. In tricultures, a capillary-like network could be demonstrated. Fluorescent Laser analysis showed crosslinked capillary-like tubes on all tested matrices. An immunohistochemical staining of CD31 confirmed that the observed capillary-like structures contained a lumen. CONCLUSIONS Prevascularisation of a tissue engineered oral mucosa was successful, implicating advantages for the clinical use. All three matrices are applicable to construct an oral mucosa equivalent for hypospadia repair. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e301 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Daniela Flesch Mainz, Germany More articles by this author Elena Frerick-Ochs Mainz, Germany More articles by this author Celine Pfeifer Mainz, Germany More articles by this author Ronald E. Unger Mainz, Germany More articles by this author Annette Schröder Mainz, Germany More articles by this author Raimund Stein Mainz, Germany More articles by this author Joachim W. Thüroff Mainz, Germany More articles by this author Walburgis Brenner Mainz, Germany More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...