Assessment of a panel of interleukin-8 reporter lung epithelial cell lines to monitor the pro-inflammatory response following zinc oxide nanoparticle exposure under different cell culture conditions

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. All cell lines were exposed to zinc oxide (ZnO) NPs at 0.6 and 6.2 μg/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). 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-α treatment. Both pIL8-GFP and pIL8-RFP also showed a marked signal induction in response to TNF-α, 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. 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. The online version of this article (doi:10.1186/s12989-015-0104-6) contains supplementary material, which is available to authorized users

Consequences of accelerated weathering in polylactide nanocomposites reinforced with halloysite nanotubes

The purpose of this study was to explore consequences of accelerated weathering on the behavior of polylactide/halloysite nanotube composites. Nanocomposites were compounded by melt mixing method via a twin-screw extruder while specimens for testing and analyses were shaped by injection molding. Behaviors of the specimens were compared before weathering and after exposure to UV irradiation and humidity steps according to Cycle-C of ISO 4892-3 standards for a total duration of 300h. IR studies revealed that photolysis and partly hydrolysis were the main degradation mechanisms leading to random main chain scission of the PLA macromolecular structure, which was reflected with drastic reductions both in the molecular weight of PLA and the mechanical properties of flexural strength and fracture toughness. On the other hand, no significant reductions were observed in the values of thermal degradation temperatures, storage modulus and flexural modulus of the specimens, which was due to the compensating effect of higher amounts of crystallinity formed during UV irradiation cycles at 70?