Improved In Vitro Model for Intranasal Mucosal Drug Delivery: Primary Olfactory and Respiratory Epithelial Cells Compared with the Permanent Nasal Cell Line RPMI 2650.

Boris Mizaikoff,Harald Luksch,Simone Ladel,Katharina Schindowski,Patrick Schlossbauer,Johannes Flamm

doi:10.3390/pharmaceutics11080367

Abstract

Background: 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. Methods: 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. Results: 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. Conclusion: 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.

Highlights

Drug delivery to the brain for the treatment of central nervous system (CNS)-related diseases became of great interest and one of the most challenging research areas in the last decade [1,2]
There are three relevant consecutive steps for nose-to-brain drug delivery (N2B): The first step is overcoming the epithelial barrier, the second step is the transport from the mucosa to sites of brain entry and the last step is the transport to other sites in the CNS [7]
We do not observe this effect as we found transepithelial electrical resistance (TEER) values around 850 Ω cm2 for respiratory epithelial primary cells (REPC) and 650 Ω cm2 for olfactory epithelial primary cells (OEPC) after 21 days of air–liquid interface (ALI) cultivation

Summary

Introduction

Drug delivery to the brain for the treatment of central nervous system (CNS)-related diseases became of great interest and one of the most challenging research areas in the last decade [1,2]. Further advantages of the intranasal application are the large surface area for absorption, rapid uptake and the avoidance of the hepatic first-pass elimination Disadvantages of this form of application are the limitation to potent drugs (only small volumes can be applied), the mucociliary clearance, and the physico-chemical features of the mucus layer (low pH, enzymatic degradation) [6,7]. The epithelial layer of the nasal mucosa is the first barrier for drug permeation during intranasal drug delivery. Porcine olfactory (OEPC) and respiratory (REPC) primary cells were characterised against the nasal tumour cell line RPMI 2650. 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

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