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. 2019 Jun 24;9:9183. doi: 10.1038/s41598-019-45137-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Barrier integrity and liposome uptake by and transport across BMEC monolayers. (A) To verify the barrier integrity of the BBB in vitro model during liposome incubation, Lipo and gH625-Lipo loaded at 5 µM PACAP (without rhodamine) were co-incubated with LY in the luminal compartment in contact with BMEC monolayers for 60 min at 37 °C. After this time, the medium of the abluminal compartment was collected and LY fluorescence was quantified by fluorimetric analysis with a spectrofluorimeter (λ_ex 430/485 nm; λ_em 535 nm). The results are expressed in permeability for LY or Pe_(LY) in 10⁻³ cm/min. Values represent means ± SD of 3 independent experiments. Note that the Pe_(LY) results show no significant impact of both liposome preparations on BMEC monolayers integrity compared to control (Ctrl: cells alone). (B) Amounts of PACAP-Rho in the luminal compartment after incubation with Lipo and gH625-Lipo loaded at 1 µM and 5 µM PACAP-Rho for 60 min at 37 °C; (C) PACAP-Rho in the BMEC lysate; (D) PACAP-Rho in the abluminal compartment. Values represent means ± SD of 5 independent experiments (**P < 0.005; ***P < 0.0005).