Shuttle‐mediated nanoparticle transport across an in vitro brain endothelium under flow conditions

Abstract

The blood brain barrier (BBB) represents a challenge in the development of new nano-delivery systems able to reach the central nervous system (CNS). In order to test the efficacy of these nanocarriers, it is fundamental to use in vitro models that resemble the in vivo cell culture conditions. Here, we demonstrate for the first time the ability of a membranotropic peptide, namely gH625, to transport a cargo-acting as a shuttle-across the BBB layer under flow conditions that mimic the blood flow rate. To this aim, a BBB microfluidic device was designed based on a transparent polyester porous membrane sandwiched between a top and a bottom overlying channel made of poly(methyl methacrylate) (PMMA). Our data clearly indicate that this microfluidic system allows the growth of brain endothelial bEnd.3 cells and the formation of a confluent layer at 7 days of culture that hinders the passage of nanoparticles compared to porous membrane alone. The device was validated at a 5 μL/min working flow rate, where the capability of the model to remain intact after nanoparticle passage was shown. Very interestingly, the decoration with the gH625 peptide enhances the adhesion of nanoparticles to the endothelial layer and the BBB crossing in flow conditions, thus confirming the efficacy of the gH625 as a delivery platform to the brain. Biotechnol. Bioeng. 2017;114: 1087-1095. © 2016 Wiley Periodicals, Inc.

L-DOPA functionalized, multi-branched gold nanoparticles as brain-targeted nano-vehicles

“…In addition, confocal reflectance imaging and transmission electron microscopy studies, coupled with the low-temperature inhibition assays, demonstrated that the majority of cell-associated AuNF were internalized in an energy-dependent process. Crucially, the amount of L-DOPA-AuNFs transported across the BBB (1.22%ID/hr) is higher than what has been reported in the literature for in vitro BBB models (0.1-1%ID/hr) 5,[33][34][35] . Furthermore, the transportation rate of AuNF across the BBB monolayer was comparable to that seen in monolayers composed of peripheral human umbilical vein endothelial cells, indicating L-DOPA-AuNF are indeed able to overcome the impermeability of the BBB.…”

Cell Penetrating Peptides as Molecular Carriers for Anti-Cancer Agents

“…The peptide gH625 (HGLASTLTRWAHYNALIRAF) was previously identified as a membrane-perturbing domain in the glycoprotein H (gH) of the herpes simplex virus type 1; gH625 interacts with model membranes, contributing to their merging and is able to traverse the membrane bilayer and transport a cargo into the cytoplasm and across the blood–brain barrier. In particular, the gH625 peptide has been shown to transport quantum dots inside the cytoplasm in an efficient way and only partially involving endocytic pathways [ 45 ].…”

Design, Synthesis and Characterization of Novel Co-Polymers Decorated with Peptides for the Selective Nanoparticle Transport across the Cerebral Endothelium

“…Among these, the gH peptide, which is derived from the glycoprotein of the Herpes simplex virus type 1, merges with model membranes and is able to traverse the membrane bilayer and to transport a cargo by escaping the degradation pathway of lysosomes [ 6 , 10 , 11 ]. In particular, in previous works, we demonstrated that the gH membranotropic peptide facilitates the delivery of polystyrene NPs across the murine BBB endothelium, which leads to a significantly higher cellular uptake by crossing through a non-conventional path [ 11 ] both in static [ 12 ] and dynamic [ 13 ] conditions. Moreover, in the case of metallic platinum ultrasmall NPs, gH partially increases their cytosolic delivery and anti-oxidant properties in Hela cells [ 14 ].…”