Inﬂuence of the fetal bovine serum proteins on the growth of human osteoblast cells on graphene Marie Kalbacova, 1 Antonin Broz, 1 Martin Kalbac 2,3 1 Institute of Inherited Metabolic Disorders, 1st Faculty of Medicine, Charles University in Prague, Ke Karlovu 2, 12852 Prague, Czech Republic 2 J. Heyrovsky Institute of Physical Chemistry, Academy of Sciences of CR, Dolejskova 3, 18223 Prague 8, Czech Republic 3 Department of Physical Chemistry, Palacky University, Olomouc 77146, Czech Republic Received 6 February 2012; revised 22 March 2012; accepted 23 April 2012 Published online in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.a.34231 Abstract: The inﬂuence of single-layer graphene produced by chemical vapor deposition on human osteoblast cells under different conditions was studied. Measurements probed the ability of cells to adhere and proliferate on graphene com- pared with SiO 2 /Si substrates and standard tissue culture plastic when cells were incubated for the ﬁrst 2 h in the pres- ence or the absence of fetal bovine serum (FBS), thus inﬂu- encing the initial, direct interaction of cells with the substrate. It was found that after 48 h of human osteoblast incubation on graphene ﬁlms, there were a comparable num- ber of cells of a similar size irrespective of the presence or the absence of serum proteins. On the other hand, a strong initial inﬂuence through the presence of FBS proteins on cell number and cell size was observed in the case of the SiO 2 /Si substrate and control plastic. Thus, our study showed that the initial presence/absence of FBS in the medium does not determine cell fate in the case of a graphene substrate, which is very unusual and different from the behavior of cells on other materials. V C 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 00A:000–000, 2012. Key Words: human osteoblast, graphene, fetal bovine serum How to cite this article: Kalbacova M, Broz A, Kalbac M. 2012. Inﬂuence of the fetal bovine serum proteins on the growth of human osteoblast cells on graphene. J Biomed Mater Res Part A 2012:00A:000–000. INTRODUCTION Typical sp 2 carbon nanomaterials, including fullerenes, car- bon nanotubes, and graphene, have many interesting physi- cal and chemical properties that are potentially useful in bio- logical and biomedical applications. 1 This is particularly the case for graphene since this material can be produced on a large scale using the chemical vapor deposition method. 2 Biocompatibility is a prerequisite for the use of gra- phene in biological or medical applications. It was recently shown that graphene has a positive effect on cell adhesion, proliferation, 3 and differentiation. 4 On the other hand, in its soluble form, graphene has also been studied for its toxicity and anticancer activity 5,6 and been found to be a good choice for such applications. As well as graphene itself, gra- phene oxide is also used as a substrate for studies of mes- enchymal stem cell differentiation, 7 showing that both sub- strates mediate different types of differentiation regarding their distinct properties. Other carbon nanostructures such as carbon nanotubes and nanocrystalline diamond have also been studied to a greater extent. 8,9 Studies on single-walled carbon nanotubes after oxygen plasma treatment showed that both surface wettability and the surface nanotopogra- phy are important factors that play a signiﬁcant role in cell cultivation. 8 It was also shown that nanocrystalline dia- monds with different termination have a strong inﬂuence on surface hydrophilic/hydrophobic characteristics and thus on cell behavior. 10 Contact between cells and substrate is mediated by focal adhesions, which are formed as multiprotein complexes. Vinculin, one of the focal adhesion proteins, is involved in the linkage between cell adhesion membranous molecules, integrins (cell receptors), and actin ﬁlaments (cell cytoskele- ton), and it plays a key role in initiating and establishing cell adhesion, the formation of cell shape, and cytoskeletal development. 11,12 The critical role of vinculin has been dem- onstrated by showing that the loss of vinculin prevents cell adhesion and spreading, stress ﬁber formation, and lamelli- podia and ﬁlopodia extension. 13 It is important to note that in most of the cell-material experiments reported so far the cells are not in direct Correspondence to: M. Kalbac; e-mail: mkalbac@hotmail.com Contract grant sponsor: Academy of Sciences of the Czech Republic; contract grant numbers: IAA400400911, KAN400100701, KAN200100801, KAN115600801. Contract grant sponsor: The Czech Grant Agency; contract grant numbers: P204/10/1677, P208/12/1062. Contract grant sponsor: Czech Ministry of Education, Youth and Sports; contract grant numbers: ME9060, MSM 0021620806. Contract grant sponsor: Charles University,1st Faculty of Medicine- project PRVOUK-P-24/LF1/3, project UNCE – 204011, Faculty of Science- project GAUK: 443211; contract grant number: SVV-2011-263206 V C 2012 WILEY PERIODICALS, INC. 1