title: Applicability of rat precision-cut lung slices in evaluating nanomaterial cytotoxicity, apoptosis, oxidative stress, and inflammation.
abstract: The applicability of rat precision-cut lung slices (PCLuS) in detecting nanomaterial (NM) toxicity to the respiratory tract was investigated evaluating sixteen OECD reference NMs (TiO₂, ZnO, CeO₂, SiO₂, Ag, multi-walled carbon nanotubes (MWCNTs)). Upon 24-hour test substance exposure, the PCLuS system was able to detect early events of NM toxicity: total protein, reduction in mitochondrial activity, caspase-3/-7 activation, glutathione depletion/increase, cytokine induction, and histopathological evaluation. Ion shedding NMS (ZnO and Ag) induced severe tissue destruction detected by the loss of total protein. Two anatase TiO₂ NMs, CeO₂ NMs, and two MWCNT caused significant (determined by trend analysis) cytotoxicity in the WST-1 assay. At non-cytotoxic concentrations, different TiO₂ NMs and one MWCNT increased GSH levels, presumably a defense response to reactive oxygen species, and these substances further induced a variety of cytokines. One of the SiO₂ NMs increased caspase-3/-7 activities at non-cytotoxic levels, and one rutile TiO₂ only induced cytokines. Investigating these effects is, however, not sufficient to predict apical effects found in vivo. Reproducibility of test substance measurements was not fully satisfactory, especially in the GSH and cytokine assays. Effects were frequently observed in negative controls pointing to tissue slice vulnerability even though prepared and handled with utmost care. Comparisons of the effects observed in the PCLuS to in vivo effects reveal some concordances for the metal oxide NMs, but less so for the MWCNT. The highest effective dosages, however, exceeded those reported for rat short-term inhalation studies. To become applicable for NM testing, the PCLuS system requires test protocol optimization.
authors.affiliation: Scientific Consultancy - Animal Welfare, Neubiberg, Germany.
authors.forename: Ursula G
authors.initials: UG
authors.lastname: Sauer
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany; Product Stewardship Water Solutions, BASF SE, Ludwigshafen, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Vogel
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Aumann
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Annemarie
authors.initials: A
authors.lastname: Hess
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Susanne N
authors.initials: SN
authors.lastname: Kolle
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Lan
authors.initials: L
authors.lastname: Ma-Hock
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany; Material Physics, BASF SE, Ludwigshafen, Germany.
authors.forename: Wendel
authors.initials: W
authors.lastname: Wohlleben
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Dammann
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Strauss
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Treumann
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Sibylle
authors.initials: S
authors.lastname: Gröters
authors.affiliation: Product Safety, BASF SE, Ludwigshafen, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Wiench
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
authors.forename: Bennard
authors.initials: B
authors.lastname: van Ravenzwaay
authors.affiliation: Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany. Electronic address: robert.landsiedel@basf.com.
authors.forename: Robert
authors.initials: R
authors.lastname: Landsiedel
mesh_terms: D023401:Animal Use Alternatives; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival; D055598:Chemical Phenomena; D003433:Crosses, Genetic; D016207:Cytokines / Q000378:metabolism; D045703:Emulsifying Agents / Q000737:chemistry; D005260:Female; D005978:Glutathione / Q000819:agonists / Q000378:metabolism; D066298:In Vitro Techniques; D008168:Lung / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008422:Materials Testing / Q000379:methods; D053768:Metal Nanoparticles / Q000737:chemistry / Q000633:toxicity / Q000648:ultrastructure; D043942:Nanotubes / Q000737:chemistry / Q000633:toxicity* / Q000648:ultrastructure; D037742:Nanotubes, Carbon / Q000737:chemistry / Q000633:toxicity / Q000648:ultrastructure; D018384:Oxidative Stress / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013010:Sonication; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D016207:Cytokines; D045703:Emulsifying Agents; D037742:Nanotubes, Carbon; D012710:Serum Albumin, Bovine; D005978:Glutathione
keywords: Cytokines; Cytotoxicity; Histopathology; Nanomaterial (NM); Oxidative stress; Rat precision-cut lung slices (PCLuS)