Impact of graphene oxide on viability of Chinese hamster ovary and mouse hepatoma MH-22A cells

Significance Statement

Advances in nanoscience and nanotechnology, enabling the synthesis of new nanomaterials, have led to the development of a number of new drug delivery systems. Carbon materials, like graphene oxide (GO), are known to be more environmentally and biologically friendly than inorganic materials, and it is one of the most common elements in our ecosystem. In biomedical applications and nanomedicine, graphene oxide have been utilized over existing drug delivery vectors due to their ability to cross cell membranes easily and their high surface area, which provides multiple attachment sites for drug targeting. Indeed, with the expanding use of graphene oxide in biomedical applications, a broader understanding of its toxicology is of high importance. The molecular basis for translocation and toxicity of graphene oxide is still largely unclear. So, for the successful application of graphene oxide in nanomedicine the knowledge of the influence of graphene oxide on the viability of various cells and the search for the ways to reduce the toxicity of graphene oxide still remains an unsolved problem. In the present work we estimated the influence of graphene oxide and graphene oxide with 10% of bovine serum albumin on Chinese hamster ovary and mouse hepatoma MH-22A cells in vitro and the structural changes of the cells by atomic force microscopy (AFM), distribution of graphene oxide inside the cells by Raman spectroscopy, imaging. Healthy and cancer cells were chosen in order to determine if the impact of graphene oxide on cell viability is different in healthy and cancer cells.

About the author

Valentinas Snitka, Prof. dr. habil.

Valentinas Snitka internationally recognized expert in nanobiotechnology. He pioneered Scanning probe microscopy research in Lithuania in 1993; visiting researcher in Campinas University, Brasil, University of Pensilvania, USA, Twente University, The Netherlands, MIMOS Corporation, Kuala Lumpur etc. His scientific expertise include Scanning Probe Microscopy methods and instrumentation, cell imaging and manipulation by SPM, electroporation of cells, nanoRaman (SERS,TERS) spectromicroscopy, synthesis and physicochemical characterization of nanoparticles, nanotoxicity, – 100+ publication in international journals, international patents, numerous invited lectures. e was serving EC as FP6 Nanotechnology, Materials and Production committee member, a member of EU Scientific and Technical Research Committee (CREST). During 2007-2008 he was appointed as Chief Scientist at MIMOS Corp. Kuala Lumpur to develop nanotechnology roadmap and infrastructure. During last 3 years he is serving EC Directorate-General for Research, Directorate G: Industrial technologies as Project Technical Advisor for implementation of FP7 projects in Nanobiotechnologies. He participates in National SF project and is a leader of project funded by European Social Fund (2013-current).

About the author

Nora Grinceviciute, M.S.

Nora Grinceviciute completed a M.S. degree in the pharmaceutical sciences program from Lithuanian University of Life Sciences. Now she is Ph.D. student at Kaunas University of Technology (Research Center for Microsystems and Nanotechnology) in chemical engineering. She has experience in industrial biotechnology (2010-2012 Sicor Biotech UAB), working with atomic force microscopy, Raman spectroscopy, Supercritical angle fluorescence microscopy, chemical functionalization, lipid bilayer formation, studying impact of nanoparticles on cells and lipid membranes. Participated in research project as younger researcher, has an international collaboration experience working in University of Zurich (Switzerland).

About the author

Danute Batiuskaite, Assoc. Prof. Dr.

Danute Batiuskaite received her PhD degree from Vytautas Magnus University, Kaunas, Lithuania, in 2003. Now she is an associate professor at the same university, the Faculty of Natural Science. She has the experience in teaching, participation in projects, national and international collaboration, doing experiments in vitro and in vivo, working with electropermeabilization of cells and tissues, searching for the optimal electrochemoterapy conditions, cytotoxicity of vitamins C and K3, impact of nanoparticles on cells.

Figure legend.

a) Raman scattering image of GO distribution in CHO cell. Intensity maps of the ‘G’ band of GO nanoparticles. b) Viability of CHO and mouse hepatoma MH-22A cells after the treatment with BSA, GO and GO–BSA at different concentrations of GO. c) AFM image of CHO control cells.

Journal Reference

Toxicol In Vitro. 2015;29(5):1195-200.

Batiuskaite D¹, Grinceviciute N², Snitka V³.

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Research Center for Microsystems and Nanotechnology, Kaunas University of Technology, 65 Studentu str., Kaunas LT-51369, Lithuania; Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, 58 K. Donelaicio str., Kaunas LT-44248, Lithuania.
Research Center for Microsystems and Nanotechnology, Kaunas University of Technology, 65 Studentu str., Kaunas LT-51369, Lithuania. Electronic address: [email protected]
Research Center for Microsystems and Nanotechnology, Kaunas University of Technology, 65 Studentu str., Kaunas LT-51369, Lithuania. Electronic address: [email protected]

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Abstract

The evaluation of the cyto- and bio-compatibility is a critical step in the development of graphene oxide (GO) as a new promising material for in vivo biomedical applications. In this study, we report the impact of GO, with and without the addition of bovine serum albumin, on healthy (Chinese hamster ovary) and a cancer (mouse hepatoma MH-22A) cells viability and the estimation of the intracellular distribution of GO inside the cells in vitro. The viability tests were performed using a colony formation assay. The intracellular distribution of GO was estimated using Raman spectroscopy and imaging. The viability of both cell lines decreased with increasing concentration of graphene oxide (12.5-50.0 μg/ml): in the case of Chinese hamster ovary cells viability decreased from 44% to 11%, in the case of mouse hepatoma MH-22A cells–from 22% to 3%. These cell lines significantly differed in their response to GO and GO-BSA formulations. The results of viability tests correlate with results of atomic force microscopy and Raman spectroscopy and imaging findings. The GO influence on cell morphology changes, cell structure, cells colony growth dynamics and GO accumulation inside the cells was higher in the case of mouse hepatoma MH-22A cells.

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