Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate

<b>Background/Objectives</b>: The purpose of this study was to create bile acid-containing liposomes to improve methotrexate blood-brain barrier penetration and to assess the liposome transportation mechanism across the blood–brain barrier. <b>Methods</b>: The improvement of...

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Main Authors: Natthan Charernsriwilaiwat, Rattanan Thaitrong, Samarwadee Plianwong, Praneet Opanasopit, Pucharee Songprakhon, Thirapit Subongkot
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Pharmaceutics
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Online Access:https://www.mdpi.com/1999-4923/17/2/269
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author Natthan Charernsriwilaiwat
Rattanan Thaitrong
Samarwadee Plianwong
Praneet Opanasopit
Pucharee Songprakhon
Thirapit Subongkot
author_facet Natthan Charernsriwilaiwat
Rattanan Thaitrong
Samarwadee Plianwong
Praneet Opanasopit
Pucharee Songprakhon
Thirapit Subongkot
author_sort Natthan Charernsriwilaiwat
collection DOAJ
description <b>Background/Objectives</b>: The purpose of this study was to create bile acid-containing liposomes to improve methotrexate blood-brain barrier penetration and to assess the liposome transportation mechanism across the blood–brain barrier. <b>Methods</b>: The improvement of liposome penetration was investigated utilizing human brain microvascular endothelial cells in an in vitro blood-brain barrier model. Using confocal laser scanning microscopy (CLSM) and flow cytometry, liposomes were labeled with fluorescent phospholipids to facilitate their passage across the blood–brain barrier. <b>Results</b>: The produced liposomes with bile acid exhibited a negative surface charge and an average particle size of between 30 and 148 nm. According to an in vitro blood-brain barrier penetration study, the methotrexate penetration was increased by liposomes containing 1% glycocholic acid but not by liposomes containing taurocholic acid. For transport pathway evaluation across the blood-brain barrier of these liposomes, CLSM revealed that fluorescent liposomes were present inside cells treated with specific endocytosis inhibitors, indicating that the cellular internalization of the particles was not involved in endocytosis. <b>Conclusions</b>: Liposomes supplemented with 1% glycocholic acid could enhance the penetration of methotrexate across the blood-brain barrier, while taurocholic acid could not. The transport of liposomes with 1% glycocholic acid across the blood-brain barrier occurs via the transcellular pathway through which it penetrates cells. In contrast, the paracellular pathway was a minor pathway.
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spelling doaj-art-454c7fcd0df24591a3a5504ee44f9e9c2025-08-20T02:01:24ZengMDPI AGPharmaceutics1999-49232025-02-0117226910.3390/pharmaceutics17020269Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of MethotrexateNatthan Charernsriwilaiwat0Rattanan Thaitrong1Samarwadee Plianwong2Praneet Opanasopit3Pucharee Songprakhon4Thirapit Subongkot5Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Burapha University, Chonburi 20131, ThailandDepartment of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Burapha University, Chonburi 20131, ThailandDepartment of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Burapha University, Chonburi 20131, ThailandDepartment of Industrial Pharmacy, Faculty of Pharmacy, Silpakorn University, Nakhon Pathom 73000, ThailandDivision of Molecular Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandDepartment of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Burapha University, Chonburi 20131, Thailand<b>Background/Objectives</b>: The purpose of this study was to create bile acid-containing liposomes to improve methotrexate blood-brain barrier penetration and to assess the liposome transportation mechanism across the blood–brain barrier. <b>Methods</b>: The improvement of liposome penetration was investigated utilizing human brain microvascular endothelial cells in an in vitro blood-brain barrier model. Using confocal laser scanning microscopy (CLSM) and flow cytometry, liposomes were labeled with fluorescent phospholipids to facilitate their passage across the blood–brain barrier. <b>Results</b>: The produced liposomes with bile acid exhibited a negative surface charge and an average particle size of between 30 and 148 nm. According to an in vitro blood-brain barrier penetration study, the methotrexate penetration was increased by liposomes containing 1% glycocholic acid but not by liposomes containing taurocholic acid. For transport pathway evaluation across the blood-brain barrier of these liposomes, CLSM revealed that fluorescent liposomes were present inside cells treated with specific endocytosis inhibitors, indicating that the cellular internalization of the particles was not involved in endocytosis. <b>Conclusions</b>: Liposomes supplemented with 1% glycocholic acid could enhance the penetration of methotrexate across the blood-brain barrier, while taurocholic acid could not. The transport of liposomes with 1% glycocholic acid across the blood-brain barrier occurs via the transcellular pathway through which it penetrates cells. In contrast, the paracellular pathway was a minor pathway.https://www.mdpi.com/1999-4923/17/2/269methotrexateliposomesbile acidsblood-brain barrier penetrationtransportation pathways
spellingShingle Natthan Charernsriwilaiwat
Rattanan Thaitrong
Samarwadee Plianwong
Praneet Opanasopit
Pucharee Songprakhon
Thirapit Subongkot
Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate
Pharmaceutics
methotrexate
liposomes
bile acids
blood-brain barrier penetration
transportation pathways
title Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate
title_full Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate
title_fullStr Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate
title_full_unstemmed Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate
title_short Development and Transportation Pathway Evaluation of Liposomes with Bile Acids for Enhancing the Blood-Brain Barrier Penetration of Methotrexate
title_sort development and transportation pathway evaluation of liposomes with bile acids for enhancing the blood brain barrier penetration of methotrexate
topic methotrexate
liposomes
bile acids
blood-brain barrier penetration
transportation pathways
url https://www.mdpi.com/1999-4923/17/2/269
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