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• W4236786214 abstract "You have accessJournal of UrologyTrauma/Reconstruction: Trauma & Reconstructive Surgery III1 Apr 2014MP9-20 PRIMARY HUMAN UROTHELIAL CELL CULTURE ON A MATRIX OF SPIDER SILK; A NOVEL TECHNIQUE IN BLADDER RECONSTRUCTION Anne Steins, Pieter Dik, Aart van Apeldoorn, Kerstin Reimers, Aart Klijn, Tom de Jong, Paul Coffer, and Koen Schepers Anne SteinsAnne Steins More articles by this author , Pieter DikPieter Dik More articles by this author , Aart van ApeldoornAart van Apeldoorn More articles by this author , Kerstin ReimersKerstin Reimers More articles by this author , Aart KlijnAart Klijn More articles by this author , Tom de JongTom de Jong More articles by this author , Paul CofferPaul Coffer More articles by this author , and Koen SchepersKoen Schepers More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.508AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Various diseases of the genitourinary tract can cause damage to the bladder which requires reconstruction. However, the ideal (bio)material for reconstruction of the bladder has not yet been discovered due to mechanical failure in vivo and other serious adverse events. A promising natural biomaterial is spider silk, which is outstanding in its slow degradation rate and mechanical properties. In this study, we assess the in vitro potential of spider silk as a suitable biomaterial for urinary tract reconstruction by cultivating primary human urothelial cells (HUCs) on native spider silk matrices. METHODS Dental wire frames were manufactured and woven with the dragline silk of female Nephila Clavipes spiders under standardized conditions. Upon sterilization of spider silk frames, HUCs were cultivated on spider silk matrices. Analysis of cell morphology and filamentous actin in HUCs showed adhesion. Expansion of HUCs was determined over time using Presto Blue and DAPI cell count in fields of vision. Viability of HUCs was determined by means of live/dead staining of HUCs cultivated on spider silk matrices and MTT assay of HUCs cultured in spider silk extract. Expression of epithelial-mesenchymal transition, adhesion, fibrosis and cellular differentiation markers was determined in HUCs at mRNA level by qRT-PCR and at protein level using flowcytometry. RESULTS Results demonstrated that spider silk supports adhesion and expansion of HUCs. In addition, spider silk had no cytotoxic effects and HUCs remained viable. QRT-PCR-based analysis revealed no major alterations in gene expression at mRNA level of HUCs cultured on spider silk. Flowcytometry-based analysis demonstrated a small subpopulation of HUCs lower in CD44 expression, indicating a slightly faster maturation of HUCs cultured on spider silk. CONCLUSIONS Together, results describe a novel biomaterial which supports adhesion, expansion and viability of HUCs. Our study gives new understanding of cellular mechanisms occurring in HUCs when cultured on spider silk and demonstrates that spider silk is a promising biomaterial for urinary tract reconstruction. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e129 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Anne Steins More articles by this author Pieter Dik More articles by this author Aart van Apeldoorn More articles by this author Kerstin Reimers More articles by this author Aart Klijn More articles by this author Tom de Jong More articles by this author Paul Coffer More articles by this author Koen Schepers More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ..." @default.
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• W4236786214 title "MP9-20 PRIMARY HUMAN UROTHELIAL CELL CULTURE ON A MATRIX OF SPIDER SILK; A NOVEL TECHNIQUE IN BLADDER RECONSTRUCTION" @default.
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