Authors :
Presenting Author: Roa'a Hamed, PhD – University of Utah
Natalia Erenburg, PhD – Hebrew University of Jerusalem
Avital Granit, PhD – Hebrew University of Jerusalem
Odelia Tepper, PhD – Hebrew University of Jerusalem
Nino Tetro, PhD – Hebrew University of Jerusalem
Erez Berman, PhD – Hebrew University of Jerusalem
Mordekhay Medvedovsky, Dr. – Hadassah Medical Center
Dana Ekstein, Dr. – Hadassah Medical Center
Sara Eyal, Professor – Hebrew University of Jerusalem
Rationale:
Cannabidiol (CBD), the most lipophilic antiseizure medication, is sequestered in body tissues. This study aimed to assess whether cellular cannabidiol sequestration alters the uptake and transmembrane transfer of small molecules.
Methods:
CBD uptake into RAW 264.7 (macrophages) and SH-SY5Y (neuroblastoma) cells was assessed using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Membrane properties were evaluated using laurdan with fluorescence microscopy. The membranal and lysosomal markers CellMask and LysoTracker, respectively, fluorescein, and the d-glucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG) served as model molecules. Transcellular transfer was evaluated in transwell systems (MDCK-MDR1 and BeWo clone b30 cell monolayers) with fluorescein and boron-dipyrromethene (BODIPY) prazosin, respectively.
Results:
CBD was detectable in RAW 264.7 cells, which slowly released it back to the medium. CBD caused shifts in membrane properties and increased cellular permeability to the membranal marker CellMask (45% increase; 95% confidence interval [CI], 124-165%; p< 0.05), fluorescein (63%; 95% CI, 43-83%; p< 0.05), and LysoTracker (up to 246% compared with other treatments, p< 0.01). However, this did not translate to altered fluorescein and BODIPY-prazosin transfer across MDCK-MDR1 cells and BeWo clone b30 cells, respectively.