824 SULPHATED GLYCOSAMINOGLYCANS (GAG'S) CONTRIBUTE TO THE BLADDER BARRIER (2012)

You have accessJournal of UrologyInfections/Inflammation of the Genitourinary Tract: Interstitial Cystitis1 Apr 2012824 SULPHATED GLYCOSAMINOGLYCANS (GAG'S) CONTRIBUTE TO THE BLADDER BARRIER Dick Janssen, Kees Jansen, Jack Schalken, and John Heesakkers Dick JanssenDick Janssen Nijmegen, Netherlands , Kees JansenKees Jansen Nijmegen, Netherlands , Jack SchalkenJack Schalken Nijmegen, Netherlands , and John HeesakkersJohn Heesakkers Nijmegen, Netherlands View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.914AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The bladder urothelium forms multiple structures that create a barrier against toxins and waste products from the urine. The glycosaminoglycan (GAG) layer is believed to be involved this barrier. The GAG-layer is located in/on the urothelium. The structure and negative charge of the GAG-layer enable it to bind H2O-molecules into a gel-like form through electrostatic entrapment. This gel forms a buffer between the urine and the urothelial cells. Examples of GAG are: chondroitin sulfate (CS), heparan sulfate (HS) and dermatan sulfate (DS). GAG-replenishing therapies are widely used for chronic bladder inflammation. However, there is a lack of scientific evidence that GAG's play an important role in the barrier of the bladder. The aim of this study is to investigate the distribution of different sulphated GAG's in the bladder and to evaluate the contribution to the bladder barrier. METHODS The location of different GAG's was investigated with GAG isolations on selective samples of the urothelium (urothelial lumen, whole urothelium) from porcine bladders (n=4) and full thickness bladder biopsies of human (n=2) and porcine bladders (n=4). After this, immunofluorescence assays on human and porcine biopsies were conducted using antibodies against HS and CS and specific GAG-digesting enzymes (chondroitinase ABC for CS and heparinase 3 for HS)and positive (protamine) and negative controls. To investigate the role of HS and CS on the urothelial barrier, the same enzymes were used on a monolayer of primary porcine urothelial cells cultured in Transwell inserts (Millipore) (HS n=8, CS n=8, control n=8). The barrier function was assessed with Trans Epithelial Electrical Resistance (TEER) on samples with TEER >500ω.cm2. RESULTS CS was detected on the luminal surface and the area just below the basal membrane of the urothelium (Fig. A, B). HS was located on the basal membrane of the urothelium (Fig. A B) and DS was located in the deeper layers of the bladder. Human and porcine bladders showed a similar distribution of GAG's. TEER measurements on the monolayer urothelial cells showed a significant decrease in barrier after CS digestion (Fig C). No effect was seen with HS digestion. CONCLUSIONS Chondroitin sulfate is localized on the luminal surface of the urothelium and plays an important role in the bladder barrier. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e336-e337 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.Metrics Author Information Dick Janssen Nijmegen, Netherlands More articles by this author Kees Jansen Nijmegen, Netherlands More articles by this author Jack Schalken Nijmegen, Netherlands More articles by this author John Heesakkers Nijmegen, Netherlands More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...