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首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >A Method for Determination and Simulation of Permeability and Diffusion in a 3D Tissue Model in a Membrane Insert System for Multi-well Plates
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A Method for Determination and Simulation of Permeability and Diffusion in a 3D Tissue Model in a Membrane Insert System for Multi-well Plates

机译：多孔板膜插入系统中3D组织模型中渗透率和扩散的确定和模拟方法

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摘要

In vitro cultivated skin models have become increasingly relevant for pharmaceutical and cosmetic applications, and are also used in drug development as well as substance testing. These models are mostly cultivated in membrane-insert systems, their permeability toward different substances being an essential factor. Typically, applied methods for determination of these parameters usually require large sample sizes (e.g., Franz diffusion cell) or laborious equipment (e.g., fluorescence recovery after photobleaching (FRAP)). This study presents a method for determining permeability coefficients directly in membrane-insert systems with diameter sizes of 4.26 mm and 12.2 mm (cultivation area). The method was validated with agarose and collagen gels as well as a collagen cell model representing skin models. The permeation processes of substances with different molecular sizes and permeation through different cell models (consisting of collagen gel, fibroblast, and HaCaT) were accurately described.Moreover, to support the above experimental method, a simulation was established. The simulation fits the experimental data well for substances with small molecular size, up to 14 x 10-10 m Stokes radius (4,000 MW), and is therefore a promising tool to describe the system. Furthermore, the simulation can considerably reduce experimental efforts and is robust enough to be extended or adapted to more complex setups.
机译：体外培养的皮肤模型与药物和化妆品应用越来越相关，并且还用于药物开发和物质测试。这些模型主要在膜插入系统中培养，它们对不同物质的渗透性是必不可少的因素。通常，用于确定这些参数的应用方法通常需要较大的样本量（例如Franz扩散池）或费力的设备（例如光漂白后的荧光恢复（FRAP））。这项研究提出了一种直接在直径为4.26mm和12.2mm（培养面积）的膜插入系统中测定渗透系数的方法。用琼脂糖和胶原蛋白凝胶以及代表皮肤模型的胶原细胞模型验证了该方法。准确地描述了不同分子大小的物质的渗透过程以及通过不同细胞模型（由胶原蛋白凝胶，成纤维细胞和HaCaT组成）的渗透过程。此外，为了支持上述实验方法，建立了一个模拟。该模拟非常适合小分子物质（最大冲程半径为14 x 10 -10 m斯托克斯（4,000 MW））的实验数据，因此是描述该系统的有前途的工具。此外，仿真可以大大减少实验工作，并且足够健壮，可以扩展或适用于更复杂的设置。

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