Department of Applied Pharmaceutics and Pharmacokinetics, Graduate School of Pharmaceutical Sciences, Kyoto University.
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University.
Biol Pharm Bull. 2022;45(9):1246-1253. doi: 10.1248/bpb.b22-00092.
Microfluidic devices are attracting attention for their ability to provide a biomimetic microenvironment wherein cells are arranged in a particular pattern and provided fluidic and mechanical forces. In this study, we evaluated drug transport across Caco-2 cell layers in microfluidic devices and investigated the effects of fluid flow on drug transport and metabolism. We designed a microfluidic device that comprises two blocks of polydimethylsiloxane and a sandwiched polyethylene terephthalate membrane with pores 3.0 µm in diameter. When cultured in a dynamic fluid environment, Caco-2 cells were multilayered and developed microvilli on the surface as compared with a static environment. Drugs with higher lipophilicity exhibited higher permeability across the Caco-2 layers, as well as in the conventional method using Transwells, and the fluidic conditions had little effect on permeability. In the Caco-2 cell layers cultured in Transwells and microfluidic devices, the basal-to-apical transport of rhodamine 123, a substrate of P-glycoprotein, was greater than the apical-to-basal transport, and the presence of tariquidar, an inhibitor of P-glycoprotein, completely diminished asymmetric transport. Furthermore, fluidic conditions promoted the metabolism of temocapril by carboxylesterases. On the other hand, we showed that fluidic conditions have little effect on gene expression of several transporters and metabolic enzymes. These results provide useful information regarding the application of microfluidic devices in drug transport and metabolism studies.
微流控装置因其能够提供仿生微环境的能力而受到关注，在这种微环境中，细胞以特定的模式排列，并提供流体和机械力。在这项研究中，我们评估了微流控装置中 Caco-2 细胞层的药物转运，并研究了流体流动对药物转运和代谢的影响。我们设计了一种微流控装置，由两块聚二甲基硅氧烷和夹在中间的聚对苯二甲酸乙二醇酯膜组成，膜上有 3.0 µm 直径的孔。与静态环境相比，在动态流体环境中培养的 Caco-2 细胞是多层的，并在表面形成微绒毛。亲脂性较高的药物在 Caco-2 层中的渗透性更高，无论是在使用 Transwells 的传统方法中，还是在流体条件下，渗透性都没有太大影响。在 Transwells 和微流控装置中培养的 Caco-2 细胞层中，罗丹明 123（P-糖蛋白的底物）从基底到顶部分泌的转运大于从顶到底部的转运，而 P-糖蛋白抑制剂 tariquidar 完全消除了不对称转运。此外，流体条件促进了 temocapril 被羧基酯酶代谢。另一方面，我们表明流体条件对几种转运体和代谢酶的基因表达影响很小。这些结果为微流控装置在药物转运和代谢研究中的应用提供了有用的信息。
