Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model

Abstract Background Targeting the blood–brain barrier (BBB) is a key step for effective brain delivery of nanocarriers. We have previously discovered that combinations of BBB nutrient transporter ligands alanine and glutathione (A-GSH), increase the permeability of vesicular and polypeptide nanocarr...

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Main Authors: Mária Mészáros, Thi Ha My Phan, Judit P. Vigh, Gergő Porkoláb, Anna Kocsis, Anikó Szecskó, Emese K. Páli, Nárcisz M. Cser, Tamás F. Polgár, Gábor Kecskeméti, Fruzsina R. Walter, Jens C. Schwamborn, Tamás Janáky, Jeng-Shiung Jan, Szilvia Veszelka, Mária A. Deli
Format: Article
Language:English
Published: BMC 2025-02-01
Series:Fluids and Barriers of the CNS
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Online Access:https://doi.org/10.1186/s12987-025-00623-2
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author Mária Mészáros
Thi Ha My Phan
Judit P. Vigh
Gergő Porkoláb
Anna Kocsis
Anikó Szecskó
Emese K. Páli
Nárcisz M. Cser
Tamás F. Polgár
Gábor Kecskeméti
Fruzsina R. Walter
Jens C. Schwamborn
Tamás Janáky
Jeng-Shiung Jan
Szilvia Veszelka
Mária A. Deli
author_facet Mária Mészáros
Thi Ha My Phan
Judit P. Vigh
Gergő Porkoláb
Anna Kocsis
Anikó Szecskó
Emese K. Páli
Nárcisz M. Cser
Tamás F. Polgár
Gábor Kecskeméti
Fruzsina R. Walter
Jens C. Schwamborn
Tamás Janáky
Jeng-Shiung Jan
Szilvia Veszelka
Mária A. Deli
author_sort Mária Mészáros
collection DOAJ
description Abstract Background Targeting the blood–brain barrier (BBB) is a key step for effective brain delivery of nanocarriers. We have previously discovered that combinations of BBB nutrient transporter ligands alanine and glutathione (A-GSH), increase the permeability of vesicular and polypeptide nanocarriers containing model cargo across the BBB. Our aim was to investigate dopamine- and ibuprofen-coupled 3-armed poly(l-glutamic acid) nanocarriers targeted by A-GSH for transfer across a novel human co-culture model with induced BBB properties. In addition, the protective effect of ibuprofen containing nanoparticles on cytokine-induced barrier damage was also measured. Method Drug-coupled nanocarriers were synthetized and characterized by dynamic light scattering and transmission electron microscopy. Cellular effects, uptake, and permeability of the nanoparticles were investigated on a human stem cell-based co-culture BBB model with improved barrier properties induced by a small molecular cocktail. The model was characterized by immunocytochemistry and permeability for marker molecules. Nanocarrier uptake in human brain endothelial cells and midbrain organoids was quantified by spectrofluorometry and visualized by confocal microscopy. The mechanisms of cellular uptake were explored by addition of free targeting ligands, endocytic and metabolic inhibitors, co-localization of nanocarriers with intracellular organs, and surface charge modification of cells. The protective effect of ibuprofen-coupled nanocarriers was investigated against cytokine-induced barrier damage by impedance and permeability measurements. Results Targeted nanoformulations of both drugs showed elevated cellular uptake in a time-dependent, active manner via endocytic mechanisms. Addition of free ligands inhibited the cellular internalization of targeted nanocarriers suggesting the crucial role of ligands in the uptake process. A higher permeability across the BBB model was measured for targeted nanocarriers. After crossing the BBB, targeted dopamine nanocarriers subsequently entered midbrain-like organoids derived from healthy and Parkinson’s disease patient-specific stem cells. The ibuprofen-coupled targeted nanocarriers showed protective effects against cytokine-induced barrier damage. Conclusion BBB-targeted polypeptide nanoparticles coupled to therapeutic molecules were effectively taken up by brain organoids or showing a BBB protective effect indicating potential applications in nervous system pathologies. Graphical Abstract
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spelling doaj-art-523645badf3a447186bcb29bed3fe0ec2025-08-20T02:15:18ZengBMCFluids and Barriers of the CNS2045-81182025-02-0122112310.1186/s12987-025-00623-2Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier modelMária Mészáros0Thi Ha My Phan1Judit P. Vigh2Gergő Porkoláb3Anna Kocsis4Anikó Szecskó5Emese K. Páli6Nárcisz M. Cser7Tamás F. Polgár8Gábor Kecskeméti9Fruzsina R. Walter10Jens C. Schwamborn11Tamás Janáky12Jeng-Shiung Jan13Szilvia Veszelka14Mária A. Deli15Institute of Biophysics, HUN-REN Biological Research CentreDepartment of Chemical Engineering, National Cheng Kung UniversityInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreDepartment of Medical Chemistry, Albert Szent-Györgyi Medical School, University of SzegedInstitute of Biophysics, HUN-REN Biological Research CentreLuxembourg Centre for Systems Biomedicine (LCSB), Developmental and Cellular Biology, University of LuxembourgDepartment of Medical Chemistry, Albert Szent-Györgyi Medical School, University of SzegedDepartment of Chemical Engineering, National Cheng Kung UniversityInstitute of Biophysics, HUN-REN Biological Research CentreInstitute of Biophysics, HUN-REN Biological Research CentreAbstract Background Targeting the blood–brain barrier (BBB) is a key step for effective brain delivery of nanocarriers. We have previously discovered that combinations of BBB nutrient transporter ligands alanine and glutathione (A-GSH), increase the permeability of vesicular and polypeptide nanocarriers containing model cargo across the BBB. Our aim was to investigate dopamine- and ibuprofen-coupled 3-armed poly(l-glutamic acid) nanocarriers targeted by A-GSH for transfer across a novel human co-culture model with induced BBB properties. In addition, the protective effect of ibuprofen containing nanoparticles on cytokine-induced barrier damage was also measured. Method Drug-coupled nanocarriers were synthetized and characterized by dynamic light scattering and transmission electron microscopy. Cellular effects, uptake, and permeability of the nanoparticles were investigated on a human stem cell-based co-culture BBB model with improved barrier properties induced by a small molecular cocktail. The model was characterized by immunocytochemistry and permeability for marker molecules. Nanocarrier uptake in human brain endothelial cells and midbrain organoids was quantified by spectrofluorometry and visualized by confocal microscopy. The mechanisms of cellular uptake were explored by addition of free targeting ligands, endocytic and metabolic inhibitors, co-localization of nanocarriers with intracellular organs, and surface charge modification of cells. The protective effect of ibuprofen-coupled nanocarriers was investigated against cytokine-induced barrier damage by impedance and permeability measurements. Results Targeted nanoformulations of both drugs showed elevated cellular uptake in a time-dependent, active manner via endocytic mechanisms. Addition of free ligands inhibited the cellular internalization of targeted nanocarriers suggesting the crucial role of ligands in the uptake process. A higher permeability across the BBB model was measured for targeted nanocarriers. After crossing the BBB, targeted dopamine nanocarriers subsequently entered midbrain-like organoids derived from healthy and Parkinson’s disease patient-specific stem cells. The ibuprofen-coupled targeted nanocarriers showed protective effects against cytokine-induced barrier damage. Conclusion BBB-targeted polypeptide nanoparticles coupled to therapeutic molecules were effectively taken up by brain organoids or showing a BBB protective effect indicating potential applications in nervous system pathologies. Graphical Abstracthttps://doi.org/10.1186/s12987-025-00623-2Blood–brain barrierIn vitro modelHuman stem cell derived endothelial cellPoly(l-glutamic acid)Dual-targeted nanocarriersAlanine
spellingShingle Mária Mészáros
Thi Ha My Phan
Judit P. Vigh
Gergő Porkoláb
Anna Kocsis
Anikó Szecskó
Emese K. Páli
Nárcisz M. Cser
Tamás F. Polgár
Gábor Kecskeméti
Fruzsina R. Walter
Jens C. Schwamborn
Tamás Janáky
Jeng-Shiung Jan
Szilvia Veszelka
Mária A. Deli
Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model
Fluids and Barriers of the CNS
Blood–brain barrier
In vitro model
Human stem cell derived endothelial cell
Poly(l-glutamic acid)
Dual-targeted nanocarriers
Alanine
title Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model
title_full Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model
title_fullStr Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model
title_full_unstemmed Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model
title_short Alanine and glutathione targeting of dopamine- or ibuprofen-coupled polypeptide nanocarriers increases both crossing and protective effects on a blood–brain barrier model
title_sort alanine and glutathione targeting of dopamine or ibuprofen coupled polypeptide nanocarriers increases both crossing and protective effects on a blood brain barrier model
topic Blood–brain barrier
In vitro model
Human stem cell derived endothelial cell
Poly(l-glutamic acid)
Dual-targeted nanocarriers
Alanine
url https://doi.org/10.1186/s12987-025-00623-2
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