title: Improving intracellular doxorubicin delivery through nanoliposomes equipped with selective tumor cell membrane permeabilizing short-chain sphingolipids.
abstract: OBJECTIVE
  To improve nanoliposomal-doxorubicin (DoxNL) delivery in tumor cells using liposome membrane-incorporated short-chain sphingolipids (SCS) with selective membrane-permeabilizing properties. DoxNL bilayers contained synthetic short-chain derivatives of known membrane microdomain-forming sphingolipids; C₈-glucosylceramide (C₈-GluCer), C₈-galactosylceramide (C₈-GalCer) or C₈-lactosylceramide (C₈-LacCer).
  
  
  METHODS
  DoxNL enriched with C₈-GluCer or C₈-GalCer were developed, optimized and characterized with regard to size, stability and drug retention. In vitro cytotoxic activity was studied in a panel of human tumor cell lines and normal cells. Intracellular Dox delivery was measured by flow cytometry and visualized by fluorescence microscopy. For a further understanding of the involved drug delivery mechanism confocal microscopy studies addressed the cellular fate of the nanoliposomes, the SCS and Dox in living cells.
  
  
  RESULTS
  C₈-LacCer-DoxNL aggregated upon Dox loading. In tumor cell lines SCS-DoxNL with C₈-GluCer or C₈-GalCer demonstrated strongly increased Dox delivery and cytotoxicity compared to standard DoxNL. Surprisingly, this effect was much less pronounced in normal cells. Nanoliposomes were not internalized, SCS however transfered from the nanoliposomal bilayer to the cell membrane and preceded cellular uptake and subsequent nuclear localization of Dox.
  
  
  CONCLUSIONS
  C₈-GluCer or C₈-GalCer incorporated in DoxNL selectively improved intracellular drug delivery upon transfer to tumor cell membranes by local enhancement of cell membrane permeability.
authors.affiliation: Laboratory Experimental Surgical Oncology Section Surgical Oncology, Department of Surgery, Erasmus Medical Center, PO Box 2040, Rotterdam 3000 CA, The Netherlands.
authors.forename: Lília R Cordeiro
authors.initials: LR
authors.lastname: Pedrosa
authors.forename: Albert
authors.initials: A
authors.lastname: van Hell
authors.forename: Regine
authors.initials: R
authors.lastname: Süss
authors.forename: Wim J
authors.initials: WJ
authors.lastname: van Blitterswijk
authors.forename: Ann L B
authors.initials: AL
authors.lastname: Seynhaeve
authors.forename: Wiggert A
authors.initials: WA
authors.lastname: van Cappellen
authors.forename: Alexander M M
authors.initials: AM
authors.lastname: Eggermont
authors.forename: Timo L M
authors.initials: TL
authors.lastname: ten Hagen
authors.forename: Marcel
authors.initials: M
authors.lastname: Verheij
authors.forename: Gerben A
authors.initials: GA
authors.lastname: Koning
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage* / Q000493:pharmacokinetics; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects*; D002463:Cell Membrane Permeability / Q000187:drug effects; D004317:Doxorubicin / Q000008:administration & dosage* / Q000493:pharmacokinetics; D005699:Galactosylceramides / Q000737:chemistry / Q000378:metabolism; D005963:Glucosylceramides / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007790:Lactosylceramides / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D009369:Neoplasms / Q000188:drug therapy; D013107:Sphingolipids / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000903:Antibiotics, Antineoplastic; D005699:Galactosylceramides; D005963:Glucosylceramides; D007790:Lactosylceramides; D008081:Liposomes; D013107:Sphingolipids; D004317:Doxorubicin