Cumulative genotoxic and apoptotic effects of xenobiotics in a mini organ culture model of human nasal mucosa as detected by the alkaline single cell microgel electrophoresis assay and the annexin V-affinity assay
作者： Michael Buehrlen , Ulrich A. Harréus , Fernando Gamarra , Rudolf Hagen , Norbert H. Kleinsasser
DOI: 10.1016/J.TOXLET.2007.01.005
关键词:
摘要: Abstract Three-dimensional mini organ cultures of human inferior nasal turbinate epithelia have proved to be a useful tool in genotoxicology studies. They allow repetitive or chronic exposure cells xenobiotics well-preserved organ-specific mucosal architecture for an extended period time. It is the aim present study concurrently monitor cumulative genotoxic and apoptotic effects sodium dichromate, N -nitrosodiethylamine (NDEA) -methyl- -nitro- -nitroso-guanidine (MNNG). Mini organs were raised by separating fresh specimens turbinates ( n = 11) into 1 mm 3 sized pieces culturing them on multiwell plates with bronchial epithelial basal medium 6 days. Aliquots subsequently exposed dichromate (1.0 mM, 1 h), NDEA (50 mM, 1 h) MNNG (0.07 mM, days 7, 9 11 versus single day only. DNA fragmentation events assessed using alkaline cell microgel electrophoresis assay (comet assay) annexin V-affinity assay. Significant could demonstrated after dichromate. Following three subsequent incubations, there was further increase genetic damage observed, accompanied rate cells. In contrast, triple incubation neither nor fraction detectable. Repetitive resulted accumulation without observable apoptosis. The results verify need assess apoptosis research investigate xenobiotics. upper aerodigestive tract shown well-suited improving ability distinguish between effects.
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