Journal
PARTICLE AND FIBRE TOXICOLOGY
Volume 12, Issue -, Pages -Publisher
BIOMED CENTRAL LTD
DOI: 10.1186/s12989-015-0104-6
Keywords
A549 cells; Interleukin-8; Air-liquid interface; Submerged cultures; Acute pulmonary (pro-)inflammatory effects
Categories
Funding
- European Union (EU) [263147]
- EU FP7 Marie Curie Actions Network for Initial Training NanoTOES [PITN-GA-2010-264506]
- Swiss National Science Foundation [64 (NRP64)]
- Adolphe Merkle Foundation
- Austrian Science Fund (FWF) [W1213] Funding Source: Austrian Science Fund (FWF)
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Background: Stably transfected lung epithelial reporter cell lines pose an advantageous alternative to replace complex experimental techniques to monitor the pro-inflammatory response following nanoparticle (NP) exposure. Previously, reporter cell lines have been used under submerged culture conditions, however, their potential usefulness in combination with air-liquid interface (ALI) exposures is currently unknown. Therefore, the aim of the present study was to compare a panel of interleukin-8 promoter (pIL8)-reporter cell lines (i.e. green or red fluorescent protein (GFP, RFP), and luciferase (Luc)), originating from A549 lung epithelial type II-like cells cells, following NPs exposure under both submerged and ALI conditions. Methods: All cell lines were exposed to zinc oxide (ZnO) NPs at 0.6 and 6.2 mu g/cm(2) for 3 and 16 hours under both submerged and ALI conditions. Following physicochemical characterization, the cytotoxic profile of the ZnO-NPs was determined for each exposure scenario. Expression of IL-8 from all cell types was analyzed at the promoter level and compared to the mRNA (qRT-PCR) and protein level (ELISA). Results: In summary, each reporter cell line detected acute pro-inflammatory effects following ZnO exposure under each condition tested. The pIL8-Luc cell line was the most sensitive in terms of reporter signal strength and onset velocity following TNF-alpha treatment. Both pIL8-GFP and pIL8-RFP also showed a marked signal induction in response to TNF-alpha, although only after 16 hrs. In terms of ZnO-NP-induced cytotoxicity pIL8-RFP cells were the most affected, whilst the pIL8-Luc were found the least responsive. Conclusions: In conclusion, the use of fluorescence-based reporter cell lines can provide a useful tool in screening the pro-inflammatory response following NP exposure in both submerged and ALI cell cultures.
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