Functionalization of liposomes with ApoE-derived peptides at different density affects cellular uptake and drug transport across a blood-brain barrier model

Cambianica, Ilaria; Zona, Cristiano; Sesana, Silvia; Gregori, Maria; Rigolio, R; Ferla, Barbara La; Nicotra, Francesco; Forloni, Gianluigi; Cagnotto, Alfredo; Salmona, Mario; Masserini, Massimo; Sancini, Giulio

doi:10.1016/j.nano.2011.05.004

Cited by 155 publications

(118 citation statements)

References 43 publications

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“…Peptide-bound NL was separated from the unbound peptide using a PD-10 column. The yield of coupling and the amount of coupled peptide was assessed by tryptophan fluorescence intensity measurements, as reported [40].…”

Section: Functionalization Of Curc-nl With Tat-peptide (Tat-curc-nl)mentioning

confidence: 99%

“…Medium was replaced and NL (100 µM) suspended in cell culture medium were incubated at 37°C with the cells for up to 48 h, corresponding to 2-fold the cell doubling time. After treatment, the cell viability was assessed by MTT (3-(4, 5-dimetiltiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay, as described [40]. Each sample was analyzed at least in triplicate.…”

Section: Assessment Of Nl Cytotoxicity On Hcmec/d3 Cellsmentioning

confidence: 99%

“…Wells were used when TEER value was higher than 40 Ω•cm 2 . Transendothelial permeability coefficient (PE) was calculated as previously described [40]. All the permeability experiments were performed in serum free culture medium at 37°C, adding 100 μM (as lipids) NL incorporating about 20000 dpm/ml of [ ]-sucrose radioactivity was counted and PE was calculated as described [40].…”

Section: Nl Uptake and Permeability Across Hcmec/d3 Cell Monolayermentioning

confidence: 99%

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Functionalization with TAT-Peptide Enhances Blood-Brain Barrier Crossing In vitro of Nanoliposomes Carrying a Curcumin-Derivative to Bind Amyloid-Β Peptide

Sancini

Gregori

Salvati

et al. 2013

J Nanomedic Nanotechnol

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# These authors contributed equally to this work preventing high-molecular weight molecules from passing through [25]. Indeed, the design and engineering of NP with high specificity for brain capillary endothelial cells have been proposed as promising strategy for AD diagnosis and treatment [26][27][28][29].The aim of the present investigation was to design NP able to bind Aβ peptide and to cross the BBB. To reach this goal we developed nanoliposomes (NL) double-functionalized with a curcuminderivative and with a modified HIV Transactivating Transcriptional Activator (TAT) peptide. Preparation of NL covalently decorated with curcumin-derivative was previously carried out, starting from a curcumin alkyne-derivative showing a very high affinity for Aβ peptide [30], suitable for NL decoration by click chemistry and with improved features of stability with respect to curcumin itself [31]. On the other side, TAT-peptide could enhance NP BBB crossing [32,33] based on the evidence that its coupling to NP may facilitate their efficient translocation through the cell membrane, bypassing the endocytic pathway [34][35][36]. TAT-peptide was covalently attached to NL surface via a thiol-maleimide reaction. The ability of NL to bind Aβ after AbstractProduction of abnormally high amounts of amyloid-β peptide in the brain plays a central role in the onset and development of Alzheimer's disease, a neurodegenerative disorder affecting millions of individuals worldwide. Nanoparticles have been proposed as promising tools to treat the disease by delivering drugs and contrast agents to the brain. Here, nanoliposomes decorated with a curcumin-derivative, displaying high affinity for amyloid-β, were functionalized with a modified cell-penetrating TAT-peptide, with the aim of conferring on such nanoliposomes the ability to cross the blood-brain barrier. Functionalization with TAT-peptide did not modify the ability of curcumindecorated nanoliposomes to bind amyloid-β fibrils, as assessed by surface plasmon resonance. Confocal microscopy, mass spectrometry and radioactivity experiments with [3 H]-sphingomyelin showed about 3-fold increase in the uptake of nanoliposomes by human brain capillary endothelial cells (hCMEC/D3) after the functionalization with TATpeptide, with no alterations in cell viability. Moreover, TAT functionalization increased the permeability of curcuminnanoliposomes across a blood-brain barrier model made with the same cells. The similar permeabilities of curcuminderivative and [3 H]-sphingomyelin suggested that nanoliposomes were transported intact. Considering these results, nanoliposomes functionalized with the curcumin-derivative and TAT-peptide represent a promising tool for targeting amyloid-β directly in the brain parenchyma.

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Section: Functionalization Of Curc-nl With Tat-peptide (Tat-curc-nl)mentioning

confidence: 99%

Section: Assessment Of Nl Cytotoxicity On Hcmec/d3 Cellsmentioning

confidence: 99%

Section: Nl Uptake and Permeability Across Hcmec/d3 Cell Monolayermentioning

confidence: 99%

See 1 more Smart Citation

Functionalization with TAT-Peptide Enhances Blood-Brain Barrier Crossing In vitro of Nanoliposomes Carrying a Curcumin-Derivative to Bind Amyloid-Β Peptide

Sancini

Gregori

Salvati

et al. 2013

J Nanomedic Nanotechnol

Self Cite

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“…For instance, the transferrin receptor is abundant in brain capillary endothelium, and it has been reported that coupling nanoparticles to transferrin can aid nanoparticle penetration into the BBB through the transferrin receptor mediated pathway. 16,24 Apolipoprotein E is an endogenous protein involved in the trafficking of naturally occurring LDL particles and when grafted on nanoparticles was reported to enhance BBB penetration, [25][26][27][28][29][30] though not necessarily translocation. 9 Similarly, cationised serum albumin has been reported to cross the BBB by absorptive-mediated trancytosis 1,31 and could also be used to gain nanoparticle access to the brain.…”

Section: Introductionmentioning

confidence: 99%

“…Indeed initial evidence suggests that the capacity of nanoparticles to cross the BBB is connected to the nature of the proteins adsorbed on their surface, such as, for instance, apolipoproteins. 15,25,26 Hence, by changing nanoparticle design, the corona that adsorbs on the nanoparticles could be used to achieve transcytosis across the BBB, and increase targeting. 39 In order to investigate the capacity of nanoparticles to cross the BBB and to test the efficiency of targeted nanoparticles to reach the brain, it is essential to be able to follow nanoparticle uptake in the BBB and to find evidence of transcytosis.…”

Section: Introductionmentioning

confidence: 99%

Nanoparticle accumulation and transcytosis in brain endothelial cell layers

et al. 2013

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The Blood-Brain Barrier (BBB) is a selective barrier, which controls and limits access to the central nervous system (CNS). The selectivity of the BBB relies on specialized characteristics of the endothelial cells that line the microvasculature, including the expression of intercellular tight junctions, which limit paracellular permeability. Several reports suggest that nanoparticles have a unique capacity to cross the BBB. However, direct evidence of nanoparticle transcytosis is difficult to obtain, and we found that typical transport studies present several limitations when applied to nanoparticles. In order to investigate the capacity of nanoparticles to access and transport across the BBB, several different nanomaterials, including silica, titania and albumin-or transferrinconjugated gold nanoparticles of different sizes, were exposed to a human in vitro BBB model of endothelial hCMEC/D3 cells. Extensive transmission electron microscopy imaging was applied in order to describe nanoparticle endocytosis and typical intracellular localisation, as well as to look for evidence of eventual transcytosis. Our results show that all of the nanoparticles were internalised, to different extents, by the BBB model and accumulated along the endo-lysosomal pathway. Rare events suggestive of nanoparticle transcytosis were also observed for several of the tested materials.

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