Models and methods to evaluate transport of drug delivery systems across cellular barriers.
Models and methods to evaluate transport of drug delivery systems across cellular barriers.
复制标题
DOI：
10.3791/50638
复制
发表时间：
2013-10-17
期刊：
影响因子：
--
通讯作者：
Muro S
中科院分区：
文献类型：
--
作者：
Ghaffarian R;Muro S
许多治疗应用需要药物载体及其货物安全有效地运输穿过体内的细胞屏障。本文描述了一种适应性的既定方法，以评估药物纳米载体（NC）穿过细胞屏障（如胃肠道（GI）上皮）的转运速率和机制。亚微米载体（纳米载体; NC）通过改善溶解度、稳定性、循环时间、靶向和释放来增强药物的功效。此外，穿过体内的细胞屏障对于将治疗性NC口服递送到循环中或从血液运输到需要干预的组织中都是至关重要的。通过以下途径实现NC跨细胞屏障的转运：（i）细胞旁途径，通过短暂破坏相邻细胞的连接，或（ii）跨细胞途径，其中物质通过内吞作用内化，跨细胞体转运，并在相对的细胞表面分泌（转胞吞）。可以通过将治疗剂或其载体与特异性结合参与转运的细胞表面标志物的靶向剂偶联来促进跨细胞屏障的递送。在这里，我们提供的方法来测量NC运输的程度和机制，通过模型细胞屏障，其中包括单层的胃肠道（GI）上皮细胞生长的多孔膜位于transwell插入。通过测量跨上皮电阻（TEER）、对照物质的跨上皮转运和紧密连接的免疫染色来确认渗透性屏障的形成。例如，使用约200 nm的聚合物NC，其携带治疗性货物并涂覆有靶向细胞表面决定簇的抗体。用125 I标记抗体或治疗性货物用于放射性同位素示踪，并将标记的NC添加到细胞单层上方的上室中不同的时间段。可以检测与细胞相关联和/或运输到下面的室的NC。游离125 I的测量允许减去降解部分。通过确定由NC转运至上述屏障参数引起的潜在变化来评估细胞旁途径。跨细胞转运是通过解决调节胞吞和胞吞途径的影响来确定的。
Many therapeutic applications require safe and efficient transport of drug carriers and their cargoes across cellular barriers in the body. This article describes an adaptation of established methods to evaluate the rate and mechanism of transport of drug nanocarriers (NCs) across cellular barriers, such as the gastrointestinal (GI) epithelium. Sub-micrometer carriers (nanocarriers; NCs) enhance efficacy of drugs by improving solubility, stability, circulation time, targeting, and release. Additionally, traversing cellular barriers in the body is crucial for both oral delivery of therapeutic NCs into the circulation or transport from the blood into tissues, where intervention is needed. NC transport across cellular barriers is achieved by: (i) the paracellular route, via transient disruption of the junctions that interlock adjacent cells, or (ii) the transcellular route, where materials are internalized by endocytosis, transported across the cell body, and secreted at the opposite cell surface (transyctosis). Delivery across cellular barriers can be facilitated by coupling therapeutics or their carriers with targeting agents that bind specifically to cell-surface markers involved in transport. Here, we provide methods to measure the extent and mechanism of NC transport across a model cell barrier, which consists of a monolayer of gastrointestinal (GI) epithelial cells grown on a porous membrane located in a transwell insert. Formation of a permeability barrier is confirmed by measuring transepithelial electrical resistance (TEER), transepithelial transport of a control substance, and immunostaining of tight junctions. As an example, ~200-nm polymer NCs are used, which carry a therapeutic cargo and are coated with an antibody that targets a cell-surface determinant. The antibody or therapeutic cargo is labeled with 125I for radioisotope tracing and labeled NCs are added to the upper chamber over the cell monolayer for varying periods of time. NCs associated to the cells and/or transported to the underlying chamber can be detected. Measurement of free 125I allows subtraction of the degraded fraction. The paracellular route is assessed by determining potential changes caused by NC transport to the barrier parameters described above. Transcellular transport is determined by addressing the effect of modulating endocytosis and transcytosis pathways.
登录
查看更多内容
用 PET 成像放射性同位素碘 124 稳定标记的聚合物纳米颗粒的内皮靶向。
影响因子：
14
作者：
Simone, Eric A.;Zern, Blaine J.;Chacko, Ann-Marie;Mikitsh, John L.;Blankemeyer, Eric R.;Muro, Silvia;Stan, Radu V.;Muzykantov, Vladimir R.
通讯作者：
Muzykantov, Vladimir R.
DOI：
10.1016/j.ejpb.2008.09.026
发表时间：
2009-03
期刊：
European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V
影响因子：
--
作者：
Torchilin V
通讯作者：
Torchilin V
DOI：
10.1016/j.jconrel.2010.10.031
发表时间：
2011-02-10
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Hsu J;Serrano D;Bhowmick T;Kumar K;Shen Y;Kuo YC;Garnacho C;Muro S
通讯作者：
Muro S
影响因子：
14.8
作者：
Hubatsch, Ina;Ragnarsson, Eva G. E.;Artursson, Per
通讯作者：
Artursson, Per
DOI：
10.1146/annurev-anchem-062011-143002
发表时间：
2012
期刊：
Annual review of analytical chemistry (Palo Alto, Calif.)
影响因子：
--
作者：
Kuhnline Sloan CD;Nandi P;Linz TH;Aldrich JV;Audus KL;Lunte SM
通讯作者：
Lunte SM