Ladel Simone, Schlossbauer Patrick, Flamm Johannes, Luksch Harald, Mizaikoff Boris, Schindowski Katharina
Institute of Applied Biotechnology, University of Applied Science Biberach, Hubertus-Liebrecht Straße 35, 88400 Biberach, Germany.
Institute of Analytical and Bioanalytical Chemistry, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
Pharmaceutics. 2019 Aug 1;11(8):367. doi: 10.3390/pharmaceutics11080367.
The epithelial layer of the nasal mucosa is the first barrier for drug permeation during intranasal drug delivery. With increasing interest for intranasal pathways, adequate in vitro models are required. Here, porcine olfactory (OEPC) and respiratory (REPC) primary cells were characterised against the nasal tumour cell line RPMI 2650.
Culture conditions for primary cells from porcine nasal mucosa were optimized and the cells characterised via light microscope, RT-PCR and immunofluorescence. Epithelial barrier function was analysed via transepithelial electrical resistance (TEER), and FITC-dextran was used as model substance for transepithelial permeation. Beating cilia necessary for mucociliary clearance were studied by immunoreactivity against acetylated tubulin.
OEPC and REPC barrier models differ in TEER, transepithelial permeation and MUC5AC levels. In contrast, RPMI 2650 displayed lower levels of MUC5AC, cilia markers and TEER, and higher FITC-dextran flux rates.
To screen pharmaceutical formulations for intranasal delivery in vitro, translational mucosal models are needed. Here, a novel and comprehensive characterisation of OEPC and REPC against RPMI 2650 is presented. The established primary models display an appropriate model for nasal mucosa with secreted MUC5AC, beating cilia and a functional epithelial barrier, which is suitable for long-term evaluation of sustained release dosage forms.
鼻黏膜上皮层是鼻腔给药时药物渗透的第一道屏障。随着对鼻腔给药途径的兴趣日益增加，需要合适的体外模型。在此，将猪嗅觉（OEPC）和呼吸（REPC）原代细胞与鼻肿瘤细胞系RPMI 2650进行了表征比较。
优化了猪鼻黏膜原代细胞的培养条件，并通过光学显微镜、逆转录聚合酶链反应（RT-PCR）和免疫荧光对细胞进行了表征。通过跨上皮电阻（TEER）分析上皮屏障功能，并使用异硫氰酸荧光素（FITC）-葡聚糖作为跨上皮渗透的模型物质。通过针对乙酰化微管蛋白的免疫反应性研究了黏液纤毛清除所必需的摆动纤毛。
OEPC和REPC屏障模型在TEER、跨上皮渗透和MUC5AC水平上存在差异。相比之下，RPMI 2650显示出较低的MUC5AC水平、纤毛标志物和TEER，以及较高的FITC-葡聚糖通量率。
为了在体外筛选鼻腔给药的药物制剂，需要转化性黏膜模型。在此，展示了针对RPMI 2650对OEPC和REPC进行的新颖而全面的表征。所建立的原代模型显示出一个适用于鼻黏膜的模型，具有分泌的MUC5AC、摆动的纤毛和功能性上皮屏障，适用于对缓释剂型进行长期评估。
