title: Expression of P-glycoprotein in excised human nasal mucosa and optimized models of RPMI 2650 cells.
abstract: To assess the transmucosal drug transport in the development of medications for intranasal administration, cellular in vitro models are preferred over the use of animal tissues due to inter-species variations and ethical concerns. With regard to the distribution of active agents and multidrug resistance, the ABC transporter P-glycoprotein plays a major role in several mammalian tissues. The present study compares the expression of this efflux pump in optimized in vitro models based on the human RPMI 2650 cell line with specimens of human turbinate mucosa. The presence of the ABCB1 gene was investigated at the mRNA and protein levels using RT-PCR and Western blot analysis in differently cultured RPMI 2650 cells and excised human nasal epithelium. Furthermore, the localization and activity of P-gp was examined by immunohistochemical staining and functionality assays using different substrates in both in vitro and ex vivo models. Both mRNA and protein expression of P-gp was found in all studied models. Furthermore, transporter functionality was detected in both RPMI 2650 cell culture models and excised human mucosa. The results demonstrated a highly promising comparability between RPMI 2650 models and explants of human nasal tissue concerning the influence of MDR1 on drug disposition. The RPMI 2650 cell line might become a useful tool in preclinical trials to improve reproducibility and achieve greater applicability to humans of experimental data regarding passive diffusion and active efflux of drug candidates.
authors.affiliation: Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Dolberg
authors.affiliation: Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany; Zentrum für Pharmaverfahrenstechnik, Technische Universität Braunschweig, Braunschweig, Germany. Electronic address: S.Reichl@tu-bs.de.
authors.forename: Stephan
authors.initials: S
authors.lastname: Reichl
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000096:biosynthesis; D002460:Cell Line; D004077:Digoxin / Q000493:pharmacokinetics; D017097:Electric Impedance; D006801:Humans; D009297:Nasal Mucosa / Q000166:cytology* / Q000378:metabolism*; D020112:Rhodamine 123 / Q000378:metabolism
chemical_list: C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020112:Rhodamine 123; D004077:Digoxin
keywords: ABC transporter; Active efflux; Drug delivery; Human nasal mucosa; In vitro model; P-glycoprotein; RPMI 2650 cell line