Al-Shehri Abdulghani, Favretto Marco E, Ioannou Panayiotis V, Romero Ignacio A, Couraud Pierre-Olivier, Weksler Babette Barbash, Parker Terry L, Kallinteri Paraskevi
School of Biomedical Sciences, University of Nottingham, Nottingham, NG7 2RD, UK.
Pharm Res. 2015 Mar;32(3):1072-83. doi: 10.1007/s11095-014-1519-8. Epub 2014 Sep 19.
Owing to restricted access of pharmacological agents into the brain due to blood brain barrier (BBB) there is a need: 1. to develop a more representative 3-D-co-culture model of tumor-BBB interaction to investigate drug and nanoparticle transport into the brain for diagnostic and therapeutic evaluation. 2. to address the lack of new alternative methods to animal testing according to replacement-reduction-refinement principles. In this work, in vitro BBB-medulloblastoma 3-D-co-culture models were established using immortalized human primary brain endothelial cells (hCMEC/D3).
hCMEC/D3 cells were cultured in presence and in absence of two human medulloblastoma cell lines on Transwell membranes. In vitro models were characterized for BBB formation, zonula occludens-1 expression and permeability to dextran. Transferrin receptors (Tfr) expressed on hCMEC/D3 were exploited to facilitate arsonoliposome (ARL) permeability through the BBB to the tumor by covalently attaching an antibody specific to human Tfr. The effect of anticancer ARLs on hCMEC/D3 was assessed.
In vitro BBB and BBB-tumor co-culture models were established successfully. BBB permeability was affected by the presence of tumor aggregates as suggested by increased permeability of ARLs. There was a 6-fold and 8-fold increase in anti-Tfr-ARL uptake into VC312R and BBB-DAOY co-culture models, respectively, compared to plain ARLs.
The three-dimensional models might be appropriate models to study the transport of various drugs and nanocarriers (liposomes and immunoarsonoliposomes) through the healthy and diseased BBB. The immunoarsonoliposomes can be potentially used as anticancer agents due to good tolerance of the in vitro BBB model to their toxic effect.
由于血脑屏障（BBB）限制了药物进入大脑，因此有必要：1. 开发一种更具代表性的肿瘤 - 血脑屏障相互作用的三维共培养模型，以研究药物和纳米颗粒进入大脑的转运情况，用于诊断和治疗评估。2. 根据替代 - 减少 - 优化原则，解决缺乏动物试验新替代方法的问题。在这项工作中，使用永生化的人原代脑内皮细胞（hCMEC/D3）建立了体外血脑屏障 - 髓母细胞瘤三维共培养模型。
hCMEC/D3细胞在Transwell膜上分别在有和没有两种人髓母细胞瘤细胞系的情况下进行培养。对体外模型进行血脑屏障形成、紧密连接蛋白 - 1表达和葡聚糖通透性的表征。利用hCMEC/D3上表达的转铁蛋白受体（Tfr），通过共价连接特异性针对人Tfr的抗体，促进砷脂质体（ARL）透过血脑屏障进入肿瘤。评估抗癌ARL对hCMEC/D3的作用。
成功建立了体外血脑屏障和血脑屏障 - 肿瘤共培养模型。如ARL通透性增加所示，肿瘤聚集体的存在影响了血脑屏障的通透性。与普通ARL相比，抗Tfr - ARL在VC312R和血脑屏障 - DAOY共培养模型中的摄取分别增加了6倍和8倍。
三维模型可能是研究各种药物和纳米载体（脂质体和免疫砷脂质体）通过健康和患病血脑屏障转运的合适模型。由于体外血脑屏障模型对其毒性作用具有良好耐受性，免疫砷脂质体有可能用作抗癌剂。
