title: Development of complex-shaped liver multicellular spheroids as a human-based model for nanoparticle toxicity assessment in vitro.
abstract: The emergence of human-based models is incontestably required for the study of complex physiological pathways and validation of reliable in vitro methods as alternative for in vivo studies in experimental animals for toxicity assessment. With this objective, we have developed and tested three dimensional environments for cells using different types of hydrogels including transglutaminase-cross-linked gelatin, collagen type I, and growth-factor depleted Matrigel. Cells grown in Matrigel exhibited the greatest cell proliferation and spheroid diameter. Moreover, analysis of urea and albumin biosynthesis revealed that the created system allowed the immortalized liver cell line HepG2 to re-establish normal hepatocyte-like properties which were not observed under the conditions of conventional cell cultures. This study presents a scalable technology for production of complex-shaped liver multicellular spheroids as a system which improves the predictive value of cell-based assays for safety and risk assessment. The time- and dose-dependent toxicity of nanoparticles demonstrates a higher cytotoxic effect when HepG2 cells grown as monolayer than embedded in hydrogels. The experimental setup provided evidence that the cell environment has significant influence on cell sensitivity and that liver spheroid is a useful and novel tool to examine nanoparticle dosing effect even at the level of in vitro studies. Therefore, this system can be applied to a wide variety of potentially hostile compounds in basic screening to provide initial warning of adverse effects and trigger subsequent analysis and remedial actions.
authors.affiliation: 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@fh-jena.de.
authors.forename: Monika
authors.initials: M
authors.lastname: Dubiak-Szepietowska
authors.affiliation: Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Karczmarczyk
authors.affiliation: Institute of Physical Chemistry, Polish Academy of Sciences, ul. Kasprzaka 44/52, 01-224 Warszawa, Poland.
authors.forename: Martin
authors.initials: M
authors.lastname: Jönsson-Niedziółka
authors.affiliation: Institute of Pharmacy, Friedrich Schiller University Jena, Semmelweissstraße 10, 07743 Jena, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Winckler
authors.affiliation: Department of Medical Engineering and Biotechnology, Ernst-Abbe-University of Applied Sciences Jena, Carl-Zeiss Promenade 2, 07745 Jena, Germany.
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Feller
mesh_terms: D000418:Albumins / Q000096:biosynthesis; D049109:Cell Proliferation; D003094:Collagen; D004338:Drug Combinations; D056945:Hep G2 Cells; D022781:Hepatocytes; D006801:Humans; D007797:Laminin; D008027:Light; D008099:Liver / Q000166:cytology* / Q000473:pathology; D008954:Models, Biological; D053758:Nanoparticles / Q000633:toxicity*; D011509:Proteoglycans; D012542:Scattering, Radiation; D018874:Spheroids, Cellular / Q000648:ultrastructure*; D014508:Urea / Q000378:metabolism
chemical_list: D000418:Albumins; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D014508:Urea; D003094:Collagen
keywords: 3D cell culture; hepatocytes; hydrogels; nanoparticles