Factors Affecting Enhanced Permeation of Amphotericin B Across Cell Membranes and Safety of Formulation.
The aim of this study was to determine amphotericin B (AmB) permeation across lipid bilayer membranes mounted on Transwell® and to observe the phagocytosis of the AmB and the AmB-lipid formulations by alveolar macrophage (AM) cell lines using a fluorescence microscope. The lipid bilayer membranes were prepared from phospholipid and ergosterol as well as phospholipid and cholesterol in a ratio (67:33 mol%). AmB-lipid formulations were prepared from AmB incorporated with four lipid derivatives during a lyophilization process. In vitro cytotoxicity studies were carried out on kidney cells by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The levels of nitric oxide production by AMs exposed to these AmB-lipid formulations were determined by the Griess reaction. Phagocytosis of the AmB-lipid formulations was carried out using AM cells. The lipid bilayer membranes and AmB-lipid formulations were successfully prepared. In vitro cytotoxicity results showed less toxicity to kidney cells than pure AmB, and a 1,000-fold less production of nitric oxide by NR8383 cell lines was obtained when compared to lipopolysaccharide. Permeation results were two- to fivefold higher than for pure AmB in the ergosterol containing lipid bilayer and two- to fourfold higher than AmB in the cholesterol containing compositions, both of which were enough to kill the fungi according to their MICs and MFCs. AM phagocytosed the AmB-lipid formulations. We suggest that these products especially the AmB-sodium deoxycholate sulfate are potential candidates for targeting AM cells for the treatment of invasive pulmonary aspergillosis.
Drug Delivery System Excellence Center, Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla University HatYai, Songkhla, 90112, Thailand.
Kajiram
K
Adhikari
Drug Delivery System Excellence Center, Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla University HatYai, Songkhla, 90112, Thailand.
Wilaiporn
W
Buatong
Drug Delivery System Excellence Center, Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla University HatYai, Songkhla, 90112, Thailand.
Ekawat
E
Thawithong
Nanotec-PSU Excellence Center on Drug Delivery System, Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla University HatYai, Songkhla, 90112, Thailand.
Tan
T
Suwandecha
Drug Delivery System Excellence Center, Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla University HatYai, Songkhla, 90112, Thailand. teerapol.s@psu.ac.th.|Nanotec-PSU Excellence Center on Drug Delivery System, Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Prince of Songkla University HatYai, Songkhla, 90112, Thailand. teerapol.s@psu.ac.th.
Teerapol
T
Srichana
D000666:Amphotericin B / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism; D003840:Deoxycholic Acid / Q000737:chemistry / Q000378:metabolism; D004338:Drug Combinations; D004875:Ergosterol / Q000737:chemistry / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D010539:Permeability; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism
D004338:Drug Combinations; D008051:Lipid Bilayers; D008055:Lipids; D010743:Phospholipids; D003840:Deoxycholic Acid; D000666:Amphotericin B; C059765:amphotericin B, deoxycholate drug combination; D002784:Cholesterol; D004875:Ergosterol
Ussing chamber; alveolar macrophage; amphotericin B; lipid bilayer; phagocytosis