I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal; FEUP - Faculdade de Engenharia, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal.
I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Rua de Jorge Viterbo Ferreira, 4050-313, Porto, Portugal.
Biomaterials. 2015 Jul;56:36-45. doi: 10.1016/j.biomaterials.2015.03.054. Epub 2015 Apr 15.
Absorption evaluation plays an increasingly important role at the early stage of drug discovery due to its potential to scan the ADME (absorption, distribution, metabolism and excretion) properties of new drug candidates. Therefore, a new three-dimensional (3D) in vitro model replicating the intestinal functioning is herein proposed aiming to dissect the stromal-epithelial interactions and evaluate the permeation of a model drug, insulin. Inspired on the intestinal mucosal architecture, the present model comprises intestinal myofibroblasts (CCD18-Co cells) embedded in Matrigel, onto which epithelial enterocytes (Caco-2 cells) and mucus-producing cells (HT29-MTX cells) were seeded. CCD18-Co myofibroblasts showed to have a central role in the remodeling of the surrounding matrix confirmed by the production of fibronectin. Subsequently, this matrix revealed to be essential to the maintenance of the model architecture by supporting the overlying epithelial cells. In terms of functionality, this model allowed the efficient prediction of insulin permeability in which the presence of mucus, the less tight character between Caco-2 and HT29-MTX epithelial cells and the 3D assembly were critical factors. Concluding, this model constitutes a robust tool in the drug development field with potential to bridge the traditional 2D cell culture models and in vivo animal models.
吸收评价在药物发现的早期阶段发挥着越来越重要的作用，因为它有可能扫描新药物候选物的 ADME（吸收、分布、代谢和排泄）特性。因此，本文提出了一种新的三维（3D）体外模型，旨在模拟肠道功能，以剖析基质-上皮相互作用并评估模型药物胰岛素的渗透。受肠道黏膜结构的启发，本模型包括嵌入 Matrigel 的肠道成纤维细胞（CCD18-Co 细胞），其上接种了肠上皮细胞（Caco-2 细胞）和产生黏液的细胞（HT29-MTX 细胞）。CCD18-Co 成纤维细胞通过产生纤维连接蛋白，在周围基质的重塑中发挥了核心作用。随后，该基质通过支持覆盖的上皮细胞，证明对模型结构的维持至关重要。就功能而言，该模型能够有效地预测胰岛素的渗透性，其中黏液的存在、Caco-2 和 HT29-MTX 上皮细胞之间不紧密的特性以及 3D 组装是关键因素。总之，该模型是药物开发领域的一个强大工具，有可能弥合传统的 2D 细胞培养模型和体内动物模型之间的差距。
