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.

IF 4.3 3区医学 Q2 CHEMISTRY, MULTIDISCIPLINARY

Pharmaceutical Research Pub Date : 2025-08-01 Epub Date: 2025-07-28 DOI:10.1007/s11095-025-03900-9

Chinenye Edith Muolokwu, Avinash Gothwal, Takahisa Kanekiyo, Jagdish Singh

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引用次数: 0

Abstract

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-T_f) achieved significantly greater transfection efficiency of pApoE2 and pGFP in neuronal cells (p < 0.0001) compared to single-modified (RDP or T_f) 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.

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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。

目的：本研究评估了双修饰脂质体在体外血脑屏障（BBB）模型中传递pApoE2和pGFP的效果，评估了它们在脑屏障神经元细胞中的转染效率，以及氢化可的松对血脑屏障完整性的影响。方法：采用脑内皮细胞（bEnd.3）与原代星形胶质细胞在transwell系统中共培养，建立体外血脑屏障模型。氢化可的松对血脑屏障完整性的影响通过经上皮电阻（TEER）、渗透性和运输研究来评估。用细胞穿透肽- rdp和转铁蛋白修饰脂质体，包封pApoE2或pgfp -壳聚糖复合物，在穿过血脑屏障后对神经元细胞进行转染。结果：与单层（p f）相比，添加150 nM HC的血脑屏障模型的TEER值显著增加，神经元细胞（p f）和普通脂质体中pApoE2和pGFP的转染效率显著提高。结论：氢化可的松增强了体外模型血脑屏障特性，使其更能代表体内血脑屏障。双修饰脂质体在血脑屏障传递遗传物质方面表现出卓越的功效，为阿尔茨海默病等神经退行性疾病的治疗干预提供了一种有希望的方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Pharmaceutical Research 医学-化学综合

CiteScore

6.60

自引率

5.40%

发文量

276

审稿时长

3.4 months

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.