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RVG29-Functionalized Lipid Nanoparticles for Quercetin Brain Delivery and Alzheimer’s Disease

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Abstract

Purpose

Lipid nanoparticles (SLN and NLC) were functionalized with the RVG29 peptide in order to target the brain and increase the neuronal uptake through the nicotinic acetylcholine receptors. These nanosystems were loaded with quercetin to take advantage of its neuroprotective properties mainly for Alzheimer’s disease.

Methods

The functionalization of nanoparticles with RVG29 peptide was confirmed by NMR and FTIR. Their morphology was assessed by transmission electron microscopy and nanoparticles size, polydispersity and zeta potential were determined by dynamic light scattering. The in vitro validation tests were conducted in hCMEC/D3 cells, a human blood-brain barrier model and thioflavin T binding assay was conducted to assess the process of amyloid-beta peptide fibrillation typical of Alzheimer’s disease.

Results

RVG29-nanoparticles displayed spherical morphology and size below 250 nm, which is compatible with brain applications. Zeta potential values were between −20 and −25 mV. Quercetin entrapment efficiency was generally higher than 80% and NLC nanoparticles were able to encapsulate up to 90%. The LDH assay showed that there is no cytotoxicity in hCMEC/D3 cell line and RVG29-nanoparticles clearly increased in 1.5-fold the permeability across the in vitro model of blood-brain barrier after 4 h of incubation compared with non-functionalized nanoparticles. Finally, this nanosystem was capable of inhibiting amyloid-beta aggregation in thioflavin T binding assay, suggesting its great potential for neuroprotection.

Conclusions

RVG29-nanoparticles that simultaneously target the blood-brain barrier and induce neurons protection against amyloid-beta fibrillation proved to be an efficient way of quercetin delivery and a promising strategy for future approaches in Alzheimer’s disease.

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Data Availability

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ACKNOWLEDGMENTS AND DISCLOSURES

This work received financial support from the European Union (FEDER funds) and National Funds (FCT/MEC, Fundação para a Ciência e a Tecnologia and Ministério da Educação e Ciência) under the Partnership Agreement PT2020 UID/QUI/50006/2013 - POCI/01/0145/FEDER/007265. Also from the UID/EQU/00511/2019—Laboratory for Process Engineering, Environment, Biotechnology and Energy—LEPABE, funded by national funds through FCT/MCTES (PIDDAC); Project POCI-01-0145-FEDER-006939, funded by FEDER funds through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI) and by national funds (PIDDAC) through FCT/MCTES;—Project “LEPABE-2-ECO-INNOVATION”—NORTE-01-0145-FEDER-000005, funded by Norte Portugal Regional Operational Programme (NORTE 2020), under PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). ARN thanks her previous Post-Doc grant under the project NORTE-01-0145-FEDER-000011. ARN also acknowledges ARDITI for her current Post-Doc grant (ARDITI-CQM_2017_011-PDG) under the project M1420-01-0145-FEDER-000005-CQM+ and the CQM strategic program PEst-OE/QUI/UI0674/2019. Andreia Granja thanks FCT for the PhD grant (SFRH/BD/130147/2017). MP thanks FCT for funding through program DL 57/2016 –Norma transitória.

The authors thank Dr. Mariana Andrade (CEMUP, UP) for technical assistance with NMR experiments and Dr. Rui Fernandes (i3S, UP) for expert help with TEM. We are also thankful to Dr. Babette Weksler from Weill Cornell Medical College (New York, USA), Dr. Ignacio A. Romero from The Open University (Milton Keynes, UK) and Dr. Pierre-Olivier Couraud from INSERM (Paris, France) for technical assistance and support with the hCMEC/D3 cell culture.

The authors declare that they have no conflict of interests.

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Authors and Affiliations

  1. LAQV, REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, 4050-313, Porto, Portugal

    R.G.R. Pinheiro, A. Granja, M. Pinheiro, A.R. Neves & S. Reis

  2. LEPABE, Departamento de Ciências da Engenharia, Faculdade de Engenharia, Universidade do Porto, 4500-465, Porto, Portugal

    J.A. Loureiro & M.C. Pereira

  3. CQM - Centro de Química da Madeira, Universidade da Madeira, Campus da Penteada, 9020-105, Funchal, Portugal

    A.R. Neves

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  1. R.G.R. Pinheiro
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  2. A. Granja
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  3. J.A. Loureiro
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  4. M.C. Pereira
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  5. M. Pinheiro
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  7. S. Reis
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Correspondence to A.R. Neves.

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Highlights

• Quercetin-loaded lipid nanoparticles functionalized with RVG29 were developed;

• No cytotoxicity of nanoparticles was detected in hCMEC/D3 cell line;

• RVG29-nanoparticles clearly increased in 1.5-fold the permeability across the BBB;

• The nanoparticles confer protection against amyloid-beta fibrillation;

• Great potential for neuroprotection in Alzheimer’s disease.

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Pinheiro, R., Granja, A., Loureiro, J. et al. RVG29-Functionalized Lipid Nanoparticles for Quercetin Brain Delivery and Alzheimer’s Disease. Pharm Res 37, 139 (2020). https://doi.org/10.1007/s11095-020-02865-1

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