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.
Part Fibre Toxicol
; 12: 29, 2015 Sep 29.
Article
en En
| MEDLINE
| ID: mdl-26415698
ABSTRACT
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 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).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-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.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.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Óxido de Zinc
/
Interleucina-8
/
Genes Reporteros
/
Nanopartículas del Metal
/
Inflamación
/
Pulmón
Límite:
Humans
Idioma:
En
Revista:
Part Fibre Toxicol
Asunto de la revista:
TOXICOLOGIA
Año:
2015
Tipo del documento:
Article
País de afiliación:
Austria