A cell-based biosensor for nanomaterials cytotoxicity assessment in three dimensional cell culture.
Nanoparticles (NPs) are widely used in consumer and medicinal products. The high prevalence of nanoparticles in the environment raises concerns regarding their effects on human health, but there is limited knowledge about how NPs interact with cells or tissues. Because the European Union has called for a substantial reduction of animal experiments for scientific purposes (Directive 2010/63), increased efforts are required to develop in vitro models to evaluate potentially hazardous agents. Here, we describe a new cell-based biosensor for the evaluation of NPs cytotoxicity. The new biosensor is based on transgenic human hepatoblastoma cells (HepG2) that express a secreted form of alkaline phosphatase (SEAP) as a reporter protein whose expression is induced upon activation of a stress response pathway controlled by the transcription regulator nuclear factor-κB (NF-κB). The NF-κB_HepG2 sensor cells were cultured in a Matrigel-based three dimensional environment to simulate the in vivo situation. The new biosensor cells offer the advantage of generating fast and reproducible readout at lower concentrations and shorter incubation time than conventional viability assays, avoid possible interaction between nanomaterials and assay compounds, therefore, minimize generation of false positive or negative results and indicate mechanism of toxicity through NF-κB signaling.
Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany. Electronic address: Monika.Dubiak-Szepietowska@eah-jena.de.
Monika
M
Dubiak-Szepietowska
Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany.
Aleksandra
A
Karczmarczyk
Institute of Pharmacy, Friedrich Schiller University Jena, Semmelweissstrasse 10, 07743 Jena, Germany.
Thomas
T
Winckler
Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany.
Karl-Heinz
KH
Feller
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay; D015374:Biosensing Techniques; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D053758:Nanoparticles / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D015398:Signal Transduction; D018675:Toxicity Tests; D014162:Transfection
D016328:NF-kappa B; D000469:Alkaline Phosphatase
3D cell culture; Cell-based biosensor; NF-κB signaling; Nanoparticles