We report a comparative investigation of the antibacterial activity of two water-soluble fullerene derivatives bearing protonated amine (AF) and deprotonated carboxylic (CF) groups appended to the fullerene cage via organic linkers. The negatively charged fullerene derivative CF showed no tendency to bind to the bacterial cells and, consequently, no significant antibacterial activity. In contrast, the compound AF loaded with cationic groups showed strong and partially irreversible binding to the negatively charged Escherichia coli K12 TG1 cells and to human erythrocytes, also possessing negative zeta potential. Adsorption of AF on the bacterial surface was visualized by atomic force microscopy revealing the formation of specific clusters (AF aggregates) surrounding the bacterial cell. Incubation of E. coli K12 TG1 with AF led to a dose-dependent bactericidal effect with ？μM. The presence of human erythrocytes in the test medium decreased the AF antibacterial activity. Thus we reveal that the water-soluble cationic fullerene derivative AF possesses promising antibacterial activity, which might be utilized in the development of novel types of chemical disinfectants. 1. Introduction Investigation of carbon-based nanomaterials (CBN) has made a great impact on the research in the fields of physics, material chemistry and technology, and also life sciences. Water-soluble forms of carbon have been studied intensively worldwide during recent decades. We would like to refer to [1], dedicated to the studies of biological activity of CBN [1]. Toxicity and biocompatibility of CBN were two major points addressed in this journal. Several review papers discussing the biological activities of CBN appeared later [2–4]. Fullerenes, the spherical carbon cages, and their functional derivatives attracted particular attention due to their unusual molecular structures and properties. Recent publications proved lower toxicity of fullerenes compared to other types of CBN, especially carbon nanotubes [5, 6]. It is known that pristine fullerenes such as C60 and C70 are very hydrophobic and possess virtually zero solubility in water. Classical fullerene derivatives bearing one organic addend appended to the fullerene cage typically have a strong tendency to aggregate in aqueous solutions [7]. Such aggregation lowers their activity significantly and hinders their practical applications. The problem can be solved by using chemically functionalized fullerene derivatives bearing a sufficient number of hydrophilic (or, even better, ionic) functional groups that significantly improve
References
[1]
R. H. Hurt, M. Monthioux, and A. Kane, “Toxicology of carbon nanomaterials: status, trends, and perspectives on the special issue,” Carbon, vol. 44, no. 6, pp. 1028–1033, 2006.
[2]
S. Bosi, T. D. Ros, G. Spalluto, and M. Prato, “Fullerene derivatives: an attractive tool for biological applications,” European Journal of Medicinal Chemistry, vol. 38, no. 11-12, pp. 913–923, 2003.
[3]
R. Brayner, “The toxicological impact of nanoparticles,” Nano Today, vol. 3, no. 1-2, pp. 48–55, 2008.
[4]
M. S. Mauter and M. Elimelech, “Environmental applications of carbon-based nanomaterials,” Environmental Science and Technology, vol. 42, no. 16, pp. 5843–5859, 2008.
[5]
C.-W. Lam, J. T. James, R. McCluskey, S. Arepalli, and R. L. Hunter, “A review of carbon nanotube toxicity and assessment of potential occupational and environmental health risks,” Critical Reviews in Toxicology, vol. 36, no. 3, pp. 189–217, 2006.
[6]
J. Kolosnjaj-Tabi, H. Szwarc, and F. Moussa, “In vivo toxicity studies of pristine carbon nanotubes: a review,” in The Delivery of Nanoparticles, A. A. Hashim, Ed., chapter 2, pp. 37–58, InTech, Vienna, Austeria, 2012.
[7]
S. Nath, H. Pal, A. V. Sapre, and J. P. Mittal, “Solvatochromism. Aggregation and photochemical properties of fullerenes, C60 and C70, in solution,” Journal of Photoscience, vol. 10, no. 1, p. 105, 2003.
[8]
F. Langa and J. -F. Nierengarten, Fullerenes: Principles and Applications, Nanoscience & Nanotechnology, Royal Society of Chemistry, Cambridge, UK, 2007.
[9]
P. A. Troshin, O. A. Troshina, R. N. Lyubovskaya, and V. F. Razumov, “Functional derivatives of fullerenes: synthesis and applications in the fields of organic electronics and biomedicine,” Ivanovo: IvSU, 2010.
[10]
S. C. Chueh, M. K. Lai, M. S. Lee, L. Y. Chiang, T. I. Ho, and S. C. Chen, “Decrease of free radical level in organ perfusate by a novel water-soluble carbon-sixty, hexa(sulfobutyl)fullerenes,” Transplantation Proceedings, vol. 31, no. 5, pp. 1976–1977, 1999.
[11]
R. Maeda-Mamiya, E. Noiri, H. Isobe et al., “In vivo gene delivery by cationic tetraamino fullerene,” Proceedings of the National Academy of Sciences of the United States of America, vol. 107, no. 12, pp. 5339–5344, 2010.
[12]
L. L. Dugan, D. M. Turetsky, C. Du et al., “Carboxyfullerenes as neuroprotective agents,” Proceedings of the National Academy of Sciences of the United States of America, vol. 94, no. 17, pp. 9434–9439, 1997.
[13]
Y. Tabata, Y. Murakami, and Y. Ikada, “Antitumor effect of poly(ethylene glycol)-modified fullerene,” Fullerene Science and Technology, vol. 5, no. 5, pp. 989–1007, 1997.
[14]
F. Kasermann and C. Kempf, “Photodynamic inactivation of enveloped viruses by buckminsterfullerene,” Antiviral Research, vol. 34, no. 1, pp. 65–70, 1997.
[15]
D. Y. Lyon, J. D. Fortner, C. M. Sayes, V. L. Colvin, and J. B. Hughes, “Bacterial cell association and antimicrobial activity of a C60 water suspension,” Environmental Toxicology and Chemistry, vol. 24, no. 11, pp. 2757–2762, 2005.
[16]
J. P. Kamat, T. P. A. Devasagayam, K. I. Priyadarsini, and H. Mohan, “Reactive oxygen species mediated membrane damage induced by fullerene derivatives and its possible biological implications,” Toxicology, vol. 155, no. 1–3, pp. 55–61, 2000.
[17]
T. Mashino, N. Usui, K. Okuda, T. Hirota, and M. Mochizuki, “Respiratory chain inhibition by fullerene derivatives: hydrogen peroxide production caused by fullerene derivatives and a respiratory chain system,” Bioorganic and Medicinal Chemistry, vol. 11, no. 7, pp. 1433–1438, 2003.
[18]
S. K. Sharma, L. Y. Chiang, and M. R. Hamblin, “Photodynamic therapy with fullerenes in vivo: reality or a dream?” Nanomedicine, vol. 6, no. 10, pp. 1813–1825, 2011.
[19]
T. Mashino, D. Nishikawa, K. Takahashi et al., “Antibacterial and antiproliferative activity of cationic fullerene derivatives,” Bioorganic and Medicinal Chemistry Letters, vol. 13, no. 24, pp. 4395–4397, 2003.
[20]
N. Tsao, P. P. Kanakamma, T.-Y. Luh, C.-K. Chou, and H.-Y. Lei, “Inhibition of Escherichia coli-induced meningitis by carboxyfullerence,” Antimicrobial Agents and Chemotherapy, vol. 43, no. 9, pp. 2273–2277, 1999.
[21]
A. A. Yurkova, E. A. Khakina, S. I. Troyanov et al., “Arbuzov chemistry with chlorofullerene C60Cl6: a powerful method for selective synthesis of highly functionalized [60]fullerene derivatives,” Chemical Communications, vol. 48, no. 71, pp. 8916–8918, 2012.
[22]
E. A. Khakina, A. A. Yurkova, A. A. Peregudov et al., “Highly selective reactions of C60Cl6 with thiols for the synthesis of functionalized [60]fullerene derivatives,” Chemical Communications, vol. 48, no. 57, pp. 7158–7160, 2012.
[23]
A. B. Kornev, E. A. Khakina, S. I. Troyanov et al., “Facile preparation of amine and amino acid adducts of [60]fullerene using chlorofullerene C60Cl6 as a precursor,” Chemical Communications, vol. 48, no. 44, pp. 5461–5463, 2012.
[24]
A. B. Kornev, A. S. Peregudov, V. M. Martynenko, J. Balzarini, B. Hoorelbeke, and P. A. Troshin, “Synthesis and antiviral activity of highly water-soluble polycarboxylic derivatives of [70]fullerene,” Chemical Communications, vol. 47, no. 29, pp. 8298–8300, 2011.
[25]
O. A. Troshina, P. A. Troshin, A. S. Peregudov, J. Kozlovskiy, and R. N. Balzarini, “Photoaddition of N-substituted piperazines to C60: an efficient approach to the synthesis of water-soluble fullerene derivatives,” Chemistry, vol. 12, no. 21, pp. 5569–5577, 2006.
[26]
O. A. Troshina, P. A. Troshin, A. S. Peregudov, V. I. Kozlovskiy, J. Balzarini, and R. N. Lyubovskaya, “Chlorofullerene C60Cl6: a precursor for straightforward preparation of highly water-soluble polycarboxylic fullerene derivatives active against HIV,” Organic and Biomolecular Chemistry, vol. 5, no. 17, pp. 2783–2791, 2007.
[27]
S. Kang, M. Herzberg, D. F. Rodrigues, and M. Elimelech, “Antibacterial effects of carbon nanotubes: size does matter!,” Langmuir, vol. 24, no. 13, pp. 6409–6413, 2008.
[28]
D. Bouchard, X. Ma, and C. Isaacson, “Colloidal properties of aqueous fullerenes: isoelectric points and aggregation kinetics of C60 and C60 derivatives,” Environmental Science & Technology, vol. 43, no. 17, pp. 6597–6603, 2009.
[29]
G. Wang, G. Zhao, and L. Yan, “Relationship between toxicity and molecular volume of dioxins, organic phosphorous compounds and n-alkanols,” Chinese Science Bulletin, vol. 49, no. 14, pp. 1437–1441, 2004.
[30]
S. S. Gayathri and A. Patnaik, “Aggregation of a C60-didodecyloxybenzene dyad: structure, dynamics, and mechanism of vesicle growth,” Langmuir, vol. 23, no. 9, pp. 4800–4808, 2007.
[31]
Y. Kanbur and Z. Kü？ükyavuz, “Synthesis and characterization of surface modified fullerene,” Fullerenes, Nanotubes and Carbon Nanostructures, vol. 20, no. 2, pp. 119–126.
[32]
X. Ma and D. Bouchard, “Formation of aqueous suspensions of fullerenes,” Environmental Science & Technology, vol. 43, no. 2, pp. 330–336, 2009.
[33]
E. Yifeng, L. Bai, L. Fan, M. Han, X. Zhang, and S. Yang, “Electrochemically generated fluorescent fullerene[60] nanoparticles as a new and viable bioimaging platform,” Journal of Materials Chemistry, vol. 21, pp. 819–823, 2011.
[34]
Y. Matsuo, K. Tahara, K. Morita, K. Matsuo, and E. Nakamura, “Regioselective eightfold and tenfold additions of a pyridine-modified organocopper reagent to [60]fullerene,” Angewandte Chemie, vol. 46, no. 16, pp. 2844–2847, 2007.
[35]
D. Y. Lyon and P. J. Alvarez, “Fullerene water suspension (nC60) exerts antibacterial effects via ROS-independent protein oxidation,” Environmental Science & Technology, vol. 42, no. 21, pp. 8127–8132, 2008.
[36]
M. HuiLi and X. J. Liang, “Fullerenes as unique nanopharmaceuticals for disease treatment,” Science China Chemistry, vol. 53, no. 11, pp. 2233–2240, 2010.
[37]
S. Liu, A. K. Ng, R. Xu et al., “Antibacterial action of dispersed single-walled carbon nanotubes on Escherichia coli and Bacillus subtilis investigated by atomic force microscopy,” Nanoscale, vol. 2, no. 12, pp. 2744–2750, 2010.
[38]
H. Schwegmann, A. J. Feitza, and F. H. Frimmela, “Influence of the zeta potential on the sorption and toxicity of iron oxide nanoparticles on S. cerevisiae and E. coli,” Journal of Colloid and Interface Science, vol. 347, no. 1, pp. 43–48, 2010.
[39]
N. Guzelsu, C. Wienstien, and S. P. Kotha, “A new streaming potential chamber for zeta potential measurements of particulates,” Review of Scientific Instruments, vol. 81, no. 1, Article ID 015106, 2010.
[40]
Y. J. Tang, J. M. Ashcroft, D. Chen et al., “Charge-associated effects of fullerene derivatives on microbial structural integrity and central metabolism,” Nano Letters, vol. 7, no. 3, pp. 754–760, 2007.
[41]
H. J. Johnston, G. R. Hutchison, F. M. Christensen, K. Aschberger, and V. Stone, “The biological mechanisms and physicochemical characteristics responsible for driving fullerene toxicity,” Toxicological Sciences, vol. 114, no. 2, pp. 162–182, 2009.
[42]
F. Rancan, S. Rosan, F. Boehm et al., “Cytotoxicity and photocytotoxicity of a dendritic C60 mono-adduct and a malonic acid C60 tris-adduct on Jurkat cells,” Journal of Photochemistry and Photobiology B, vol. 67, no. 3, pp. 157–162, 2002.
[43]
J. Wong-Ekkabut, S. Baoukina, W. Triampo, I.-M. Tang, D. P. Tieleman, and L. Monticelli, “Computer simulation study of fullerene translocation through lipid membranes,” Nature Nanotechnology, vol. 3, no. 6, pp. 363–368, 2008.
[44]
I. Nakanishi, S. Fukuzumi, T. Konishi et al., “Fullerenes for the new millennium,” in Proceedings of the International Symposium on Fullerenes, Nanotubes, and Carbon Nanoclusters, P. V. Kamat, D. M. Guldi, and K. M. Kadish, Eds., vol. 11, The Electrochemical Society, Pennington, NJ, USA, 2001.
[45]
M. Carini, L. Djordjevic, and T. D. Ros, “Fullerenes in biology and medicine,” in Handbook of Carbon Nano Materials, vol. 3, pp. 1–48, World Scientific Publishing, 2012.
[46]
A. Aquino, J. Chan, K. Giolma, and M. Loh, “The effect of a fullerene water suspension on the growth, cell viability, and membrane integrity of Escherichia coli B23,” Journal of Eexperimental Microbiology & Immunology, vol. 14, pp. 13–20, 2010.
[47]
T. Mashino, K. Okuda, T. Hirota, M. Hirobe, T. Nagano, and M. Mochizuki, “Inhibition of E. coli growth by fullerene derivatives and inhibition mechanism,” Bioorganic and Medicinal Chemistry Letters, vol. 9, no. 20, pp. 2959–2962, 1999.
[48]
G. P. Tegos, T. N. Demidova, D. Arcila-Lopez et al., “Cationic fullerenes are effective and selective antimicrobial photosensitizers,” Chemistry and Biology, vol. 12, no. 10, pp. 1127–1135, 2005.
