{"title":"双重修饰脂质体包裹核酸（pApoE2或pGFP）在氢化可的松增强的体外血脑屏障模型中的转运研究，用于中枢神经系统治疗筛选。","authors":"Chinenye Edith Muolokwu, Avinash Gothwal, Takahisa Kanekiyo, Jagdish Singh","doi":"10.1007/s11095-025-03900-9","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>The study assessed dual-modified liposomes for delivering pApoE2 and pGFP across an in vitro blood-brain barrier (BBB) model supplemented with hydrocortisone (HC), evaluating their transfection efficiency in neuronal cells across the BBB and the impact of hydrocortisone on BBB integrity.</p><p><strong>Methods: </strong>An in vitro BBB model was developed using brain endothelial cells (bEnd.3) co-cultured with primary astrocytes in a transwell system. Hydrocortisone's effect on BBB integrity was assessed via transepithelial electrical resistance (TEER), permeability and transport studies. Liposomes, modified with cell-penetrating peptide-RDP and Transferrin, encapsulating pApoE2 or pGFP-chitosan polyplex, were evaluated for neuronal cell transfection after crossing the BBB.</p><p><strong>Results: </strong>The BBB models supplemented with 150 nM HC showed a significant increase in TEER values compared to monolayers (p < 0.0001) and co-culture BBB models without HC supplementation (p < 0.01), indicating enhanced BBB integrity. Permeability assays demonstrated reduced sodium fluorescein translocation across the 150 nM hydrocortisone-supplemented BBB models compared to monolayers (p < 0.001) and co-culture models without HC supplementation (p < 0.05). Liposomes exhibited good characteristics and efficient encapsulation of pApoE2 or pGFP-chitosan polyplex, and successfully crossed the developed BBB model. Dual-modified liposomes (RDP-T<sub>f</sub>) achieved significantly greater transfection efficiency of pApoE2 and pGFP in neuronal cells (p < 0.0001) compared to single-modified (RDP or T<sub>f</sub>) and plain liposomes.</p><p><strong>Conclusions: </strong>Hydrocortisone enhanced the BBB properties of the in vitro model, making it more representative of the in vivo BBB. Dual-modified liposomes demonstrated superior efficacy in delivering genetic materials across the BBB, providing a promising approach for therapeutic interventions in neurodegenerative diseases like Alzheimer's.</p>","PeriodicalId":20027,"journal":{"name":"Pharmaceutical Research","volume":" ","pages":"1331-1345"},"PeriodicalIF":4.3000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Dual-modified Liposomes Encapsulating Nucleic Acids (pApoE2 or pGFP) for Transport Studies Across a Hydrocortisone-enhanced In Vitro Blood-brain Barrier Model for CNS Therapeutic Screening.\",\"authors\":\"Chinenye Edith Muolokwu, Avinash Gothwal, Takahisa Kanekiyo, Jagdish Singh\",\"doi\":\"10.1007/s11095-025-03900-9\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>The study assessed dual-modified liposomes for delivering pApoE2 and pGFP across an in vitro blood-brain barrier (BBB) model supplemented with hydrocortisone (HC), evaluating their transfection efficiency in neuronal cells across the BBB and the impact of hydrocortisone on BBB integrity.</p><p><strong>Methods: </strong>An in vitro BBB model was developed using brain endothelial cells (bEnd.3) co-cultured with primary astrocytes in a transwell system. Hydrocortisone's effect on BBB integrity was assessed via transepithelial electrical resistance (TEER), permeability and transport studies. Liposomes, modified with cell-penetrating peptide-RDP and Transferrin, encapsulating pApoE2 or pGFP-chitosan polyplex, were evaluated for neuronal cell transfection after crossing the BBB.</p><p><strong>Results: </strong>The BBB models supplemented with 150 nM HC showed a significant increase in TEER values compared to monolayers (p < 0.0001) and co-culture BBB models without HC supplementation (p < 0.01), indicating enhanced BBB integrity. Permeability assays demonstrated reduced sodium fluorescein translocation across the 150 nM hydrocortisone-supplemented BBB models compared to monolayers (p < 0.001) and co-culture models without HC supplementation (p < 0.05). Liposomes exhibited good characteristics and efficient encapsulation of pApoE2 or pGFP-chitosan polyplex, and successfully crossed the developed BBB model. Dual-modified liposomes (RDP-T<sub>f</sub>) achieved significantly greater transfection efficiency of pApoE2 and pGFP in neuronal cells (p < 0.0001) compared to single-modified (RDP or T<sub>f</sub>) and plain liposomes.</p><p><strong>Conclusions: </strong>Hydrocortisone enhanced the BBB properties of the in vitro model, making it more representative of the in vivo BBB. Dual-modified liposomes demonstrated superior efficacy in delivering genetic materials across the BBB, providing a promising approach for therapeutic interventions in neurodegenerative diseases like Alzheimer's.</p>\",\"PeriodicalId\":20027,\"journal\":{\"name\":\"Pharmaceutical Research\",\"volume\":\" \",\"pages\":\"1331-1345\"},\"PeriodicalIF\":4.3000,\"publicationDate\":\"2025-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Pharmaceutical Research\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1007/s11095-025-03900-9\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/7/28 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Pharmaceutical Research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s11095-025-03900-9","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/7/28 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
Dual-modified Liposomes Encapsulating Nucleic Acids (pApoE2 or pGFP) for Transport Studies Across a Hydrocortisone-enhanced In Vitro Blood-brain Barrier Model for CNS Therapeutic Screening.
Purpose: The study assessed dual-modified liposomes for delivering pApoE2 and pGFP across an in vitro blood-brain barrier (BBB) model supplemented with hydrocortisone (HC), evaluating their transfection efficiency in neuronal cells across the BBB and the impact of hydrocortisone on BBB integrity.
Methods: An in vitro BBB model was developed using brain endothelial cells (bEnd.3) co-cultured with primary astrocytes in a transwell system. Hydrocortisone's effect on BBB integrity was assessed via transepithelial electrical resistance (TEER), permeability and transport studies. Liposomes, modified with cell-penetrating peptide-RDP and Transferrin, encapsulating pApoE2 or pGFP-chitosan polyplex, were evaluated for neuronal cell transfection after crossing the BBB.
Results: The BBB models supplemented with 150 nM HC showed a significant increase in TEER values compared to monolayers (p < 0.0001) and co-culture BBB models without HC supplementation (p < 0.01), indicating enhanced BBB integrity. Permeability assays demonstrated reduced sodium fluorescein translocation across the 150 nM hydrocortisone-supplemented BBB models compared to monolayers (p < 0.001) and co-culture models without HC supplementation (p < 0.05). Liposomes exhibited good characteristics and efficient encapsulation of pApoE2 or pGFP-chitosan polyplex, and successfully crossed the developed BBB model. Dual-modified liposomes (RDP-Tf) achieved significantly greater transfection efficiency of pApoE2 and pGFP in neuronal cells (p < 0.0001) compared to single-modified (RDP or Tf) and plain liposomes.
Conclusions: Hydrocortisone enhanced the BBB properties of the in vitro model, making it more representative of the in vivo BBB. Dual-modified liposomes demonstrated superior efficacy in delivering genetic materials across the BBB, providing a promising approach for therapeutic interventions in neurodegenerative diseases like Alzheimer's.
期刊介绍：
Pharmaceutical Research, an official journal of the American Association of Pharmaceutical Scientists, is committed to publishing novel research that is mechanism-based, hypothesis-driven and addresses significant issues in drug discovery, development and regulation. Current areas of interest include, but are not limited to:
-(pre)formulation engineering and processing-
computational biopharmaceutics-
drug delivery and targeting-
molecular biopharmaceutics and drug disposition (including cellular and molecular pharmacology)-
pharmacokinetics, pharmacodynamics and pharmacogenetics.
Research may involve nonclinical and clinical studies, and utilize both in vitro and in vivo approaches. Studies on small drug molecules, pharmaceutical solid materials (including biomaterials, polymers and nanoparticles) biotechnology products (including genes, peptides, proteins and vaccines), and genetically engineered cells are welcome.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
