Ethanol

Scanning electron microscopy was used to visualize MSSA due to its ability to potentially exist within the patterned surface. After bacterial immersion, two samples of each the MP and two smooth samples were retained for analysis without RODAC exposure, while another two samples of each surface were stamped with a RODAC plate. Each sample was subsequently fixed with osmium tetroxide gas (Electron Microscopy Sciences, 19150) for 45 m and then subjected to a dehydration series with 10 m incubations in 25, 50, 75, and finally 95% ethanol (Decon Labs Inc, King of Prussia, PA), then air-dried overnight at ambient conditions. Two 8 mm circles were taken near the center of each sample with a biopsy punch (Fisher, Waltham, MA), mounted, sputter-coated with gold, and analyzed via SEM. Two images were taken per smooth sample and four images were taken per the MP sample for qualitative image analysis.

The stock solution of ThT was prepared by dissolving ThT (Sigma–Aldrich) in ethanol (Decon Labs) and filtering through 0.22 mm polyethersulfone syringe filter. To monitor the fibril formation, Aβ40 peptides were diluted to 50 μM in fibril growth buffer in a black 96-well plate with clear bottom. For control experiment, ammonium acetate solution in a volume equivalent to the peptides volume was added to the growth buffer instead of Aβ40 peptides. To monitor the effect of bexarotene on Aβ40 fibrillization, bexarotene was added to diluted Aβ40 solution at final desired concentrations. ThT then was added to the Aβ40, Aβ40-bexarotene, and control solutions to a final concentration of 35 μM. The ThT fluorescence signal was measured at 37 ^oC (with mixing every 5 min) every 15 min with excitation and emission at 442 and 488 nm, respectively, for a period of 15 h using SpectraMax Gemini EM Microplate Reader (Molecular Devices). The sigmoid curves were obtained by subtracting the absorbance of the control and normalizing all the data points to their maximum value.

For the drinking studies, mice were offered 20% ethanol (v/v, in tap water; 200 proof, DeCon Labs, King of Prussia, PA, USA). For injections, 20% ethanol (v/v, in 0.9% saline) was administered interperitoneally (IP).