Membrane permeation of giant unilamellar vesicles and corneal epithelial cells with lipophilic vitamin nanoemulsions.

Nanoemulsions of a lipophilic vitamin, retinol palmitate (vitamin A; VA), have a therapeutic effect on corneal damage. The nanoemulsion based on a triblock-type polymer surfactant with polyoxyethylene and polypropylene, EO100PO70EO100 (EOPO) showed superior efficacy, as compared with a nanoemulsion based on polyoxyethylene (60) hydrogenated castor oil (HCO). We studied the mechanism of VA nanoemulsions related to efficacy from the viewpoint of the interaction with plasma membrane-mimicking giant unilamellar vesicles (GUVs) and the plasma membrane permeation in corneal epithelial cells. When nanoemulsions and GUVs doped with fluorescent compounds were mixed each other, and observed by confocal laser microscopy, EOPO nanoemulsions induced endocytic morphological changes like strings and vesicles of the bilayer drawn inside a GUV by budding. Judging by isothermal titration calorimetry and ζ potential measurements, the EOPO nanoemulsions seemed to have stronger hydrophobic interactions with the lipid bilayer because of lower coverage of the core interface. Next, when the nanoemulsions prepared with a pyrene derivative of retinol (VApyr) were applied to corneal epithelial cells, the EOPO nanoemulsions greatly permeated the cells and gathered around the cell nucleus, as compared with HCO nanoemulsions. Furthermore, according to the three-dimensional images of the cell, it was found that the vesicles that absorbed nanoemulsions formed from the plasma membrane as real endocytosis, and were transported to the area around the nucleus. Consequently, it is likely that EOPO nanoemulsions entered the cell by membrane-mediated transport, delivering VA to the cell nucleus effectively and enhancing the effects of VA.

Xylarianaphthol-1, a novel dinaphthofuran derivative, activates p21 promoter in a p53-independent manner.

Xylarianaphthol-1, a novel dinaphthofuran derivative, was isolated from a marine sponge-derived fungus of order Xylariales on the guidance of a bioassay using the transfected human osteosarcoma MG63 cells (MG63(luc+)). The chemical structure of xylarianaphthol-1 was determined from the (1)H and (13)C NMR analysis and was further confirmed by the total synthesis. Xylarianaphthol-1 activated p21 promoter stably transfected in MG63 cells dose-dependently. Expression of p21 protein in the wild-type MG63 cells was also increased by xylarianaphthol-1 treatment.