36 ENDOCYTOSIS UPTAKE OF LIPOSOMES IN UROTHELIUM CELLS DETECTED BY TRANSMISSION ELECTRON MICROSCOPY (2012)

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Basic Research I1 Apr 201236 ENDOCYTOSIS UPTAKE OF LIPOSOMES IN UROTHELIUM CELLS DETECTED BY TRANSMISSION ELECTRON MICROSCOPY Jayabalan Nirmal, Pradeep Tyagi, Loan Dang, Ann Hanna-Mitchell, Amanda Wolf-Johnston, Jonathan Kaufman, Lori Birder, and Michael Chancellor Jayabalan NirmalJayabalan Nirmal Royal Oak, MI More articles by this author , Pradeep TyagiPradeep Tyagi Royal Oak, MI More articles by this author , Loan DangLoan Dang Rochester, MI More articles by this author , Ann Hanna-MitchellAnn Hanna-Mitchell Pittsburgh, PA More articles by this author , Amanda Wolf-JohnstonAmanda Wolf-Johnston Pittsburgh, PA More articles by this author , Jonathan KaufmanJonathan Kaufman Pittsburgh, PA More articles by this author , Lori BirderLori Birder Pittsburgh, PA More articles by this author , and Michael ChancellorMichael Chancellor Royal Oak, MI More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.079AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Intravesical liposomes (phospholipid membrane vesicles) have been used as a drug delivery platform and more recently, empty liposomes has shown promise as a treatment of interstitial cystitis/painful bladder syndrome. The exact mechanism for liposome uptake in bladder cells remains to be investigated and in the present study, we investigated the role of endocytosis in uptake of liposomes by urothelium cells. METHODS Urothelial cell line (UROtsa) were left to adhere and grow for 48h in 100mm petridishes using Dulbecco modified Eagle medium (DMEM) containing 5% v/v fetal bovine serum with incubation at 37°C in a 5% CO2:95% air atmosphere. Liposomes encapsulating colloidal gold particles size 10nm were prepared by thin film hydration method and were incubated with serum free UROtsa cells for 2h either at 37°C or at 4°C. The morphology of liposome–cell interactions at two temperatures was assessed by transmission electron microscopy. RESULTS Transmission Electron Microscopy (TEM) images located endocytic vesicles containing liposomes inside the UROtsa cells incubated at 37°C [panel B of attached picture] as indicated by dark stain of colloidal gold particles 10nm. In contrast, only extracellular binding was noticed in cells incubated at 4°C[panel A of attached picture]. Thin section electron micrographs show gold–containing liposomes trapped in intracellular vesicles, and in secondary lysosomes incubated at 37°C with colloidal gold liposomes. Absence of liposome internalization at 4°C indicate the need of energy dependent endocytosis as the primary mechanism of entry of liposomes into urothelium. CONCLUSIONS Based on these observations, we propose that the main route of entry of liposomes into urothelium cells is by endocytosis and ultrastructural study provide evidence for a possible mechanism by which liposomes can act as drug delivery platform. These findings support the usefulness of liposomes in intravesical drug delivery. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetailsCited byKashyap M, Kawamorita N, Tyagi V, Sugino Y, Chancellor M, Yoshimura N and Tyagi P (2018) Down-Regulation of Nerve Growth Factor Expression in the Bladder by Antisense Oligonucleotides as New Treatment for Overactive BladderJournal of Urology, VOL. 190, NO. 2, (757-764), Online publication date: 1-Aug-2013. Volume 187Issue 4SApril 2012Page: e15 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jayabalan Nirmal Royal Oak, MI More articles by this author Pradeep Tyagi Royal Oak, MI More articles by this author Loan Dang Rochester, MI More articles by this author Ann Hanna-Mitchell Pittsburgh, PA More articles by this author Amanda Wolf-Johnston Pittsburgh, PA More articles by this author Jonathan Kaufman Pittsburgh, PA More articles by this author Lori Birder Pittsburgh, PA More articles by this author Michael Chancellor Royal Oak, MI More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...