title: In-vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix-based human nasal primary culture systems for nasal drug delivery studies.
abstract: The aim of this study was to establish a collagen matrix-based nasal primary culture system for drug delivery studies. Nasal epithelial cells were cultured on derivatised (Cellagen membrane CD-24), polymerised (Vitrogen gel) and fibrillar (Vitrogen film) collagen substrata. Cell morphology was assessed by microscopy. The cells were further characterised by measurement of ciliary beat frequency (CBF), transepithelial resistance (TER), permeation of sodium fluorescein, mitochondrial dehydrogenase (MDH) activity and lactate dehydrogenase (LDH) release upon cell exposure to sodium tauro-24, 25 dihydrofusidate (STDHF). Among the three collagen substrata investigated, the best epithelial differentiated phenotype (monolayer with columnar/cuboidal morphology) occurred in cells grown on Cellagen membrane CD-24 between day 4 and day 11. Cell culture reproducibility was better with Cellagen membrane CD-24 (90%) in comparison with Vitrogen gel (70%) and Vitrogen film (< 10%). TER was higher in cells grown on Vitrogen gel than on Cellagen membrane CD-24 and Vitrogen film. The apparent permeability coefficient (Papp x 10(-7)cm s(-1)) of sodium fluorescein in these conditions was 0.45+/-0.08 (Vitrogen gel) and 1.91+/-0.00 (Cellagen membrane CD-24). Except for LDH release, CBF and cell viability were comparable for all the substrata. Based on MDH activity, LDH release, CBF, TER and permeation studies, Cellagen membrane CD-24- and Vitrogen gel-based cells were concluded to be functionally suitable for in-vitro nasal drug studies. Vitrogen film-based cultures may be limited to metabolism and cilio-toxicity studies.
authors.affiliation: Laboratorium voor Farmacotechnologie en Biofarmacie, K. U. Leuven, Belgium.
authors.forename: R U
authors.initials: RU
authors.lastname: Agu
authors.forename: M
authors.initials: M
authors.lastname: Jorissen
authors.forename: T
authors.initials: T
authors.lastname: Willems
authors.forename: P
authors.initials: P
authors.lastname: Augustijns
authors.forename: R
authors.initials: R
authors.lastname: Kinget
authors.forename: N
authors.initials: N
authors.lastname: Verbeke
mesh_terms: D000281:Administration, Intranasal; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002923:Cilia / Q000502:physiology / Q000648:ultrastructure; D003094:Collagen / Q000378:metabolism / Q000494:pharmacology*; D016503:Drug Delivery Systems; D019793:Fluorescein / Q000378:metabolism; D005672:Fusidic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000187:drug effects / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000187:drug effects / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D009297:Nasal Mucosa / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D005672:Fusidic Acid; C053824:tauro-24,25-dihydrofusidate; D003094:Collagen; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase; D019793:Fluorescein