Abstract
Fullerene is a compound formed during carbon burst that has been produced synthetically starting from the 1990s. The spherical shape and the characteristic carbon bonds of this allotrope (C60) have made it a suitable molecule for many applications. During the last decade, the low aqueous solubility of this molecule has been improved by chemical functionalization allowing the use of fullerene derivatives in biological fluids. The characterization of the toxicity potential of fullerenes is therefore of growing interest for any biomedical application. Intravenous injection is one of the possible routes of their administrations and therefore red blood cells are among the first targets of fullerene cytotoxicity. Human red blood cells are easily available and separated from plasma. Membrane disruption by toxic compounds is easily detected in red blood cells as release of hemoglobin in the cell medium, which can be assayed spectrophotometrically at λ = 415 nm. Due to the high molar extinction coefficient of hemoglobin, the assay can be performed on a small amount of both red blood cells and the test compounds, which might be available only in small quantities. So, the hemolysis assay is a simple screening test, whose results can guide further investigations on cytotoxicity in more complex experimental models.
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References
Prato M (1997) [60]Fullerene chemistry for materials science applications. J Mater Chem 7:1097–1109
Beuerle F, Hirsch A (2009) Synthesis and orthogonal functionalization of [60]fullerene e, e, e-trisadducts with two spherically defined addend zones. Chemistry 15:7434–7446
Bianco A, Da Ros T, Prato M, Toniolo C (2001) Fullerene-based amino acids and peptides. J Pept Sci 4:208–219
Jagadeesan D, Eswaramoorthy M (2010) Functionalized carbon nanomaterials derived from carbohydrates. Chem Asian J 5:232–243
Nakamura E, Isobe H (2003) Functionalized fullerenes in water. The first 10 years of their chemistry, biology, and nanoscience. Acc Chem Res 36:807–815
Witte P, Beuerle F, Hartnagel U, Lebovitz R, Savouchkina A, Sali S, Guldi D, Chronakis N, Hirsch A (2007) Water solubility, antioxidant activity and cytochrome C binding of four families of exohedral adducts of C60 and C70. Org Biomol Chem 5:3599–3613
Gharbi N, Pressac M, Hadchouel M, Szwarc H, Wilson SR, Moussa F (2005) [60]Fullerene is a powerful antioxidant in vivo with no acute or subacute toxicity. Nano Lett 5:2578–2585
Sun T, Xu Z (2006) Radical scavenging activities of alpha-alanine C60 adduct. Bioorg Med Chem Lett 16:3731–3734
Enes RF, Tomé AC, Cavaleiro JA, Amorati R, Fumo MG, Pedulli GF, Valgimigli L (2006) Synthesis and antioxidant activity of [60]fullerene-BHT conjugates. Chemistry 12:4646–4653
Hu Z, Guan W, Wang W, Huang L, Xing H, Zhu Z (2007) Synthesis of β-alanine C60 derivative and its protective effect on hydrogen peroxide-induced apoptosis in rat pheochromocytoma cells. Cell Biol Int 31:798–804
Yang J, Alemany LB, Driver J, Hartgerink JD, Barron AR (2007) Fullerene-derivatized amino acids: Synthesis, characterization, antioxidant properties, and solid-phase peptide synthesis. Chem Eur J 13:2530–2545
Dugan L, Gabrielsen J, Yu S, Lin T, Choi D (1996) Buckminsterfullerenol free radical scavengers reduce excitotoxic and apoptotic death of cultured cortical neurons. Neurobiol Dis 3:129–135
Dugan LL, Turetsky DM, Du C, Lobner D, Wheeler M, Almli CR, Shen CKF, Luh TY, Choi DW, Lin TS (1997) Carboxyfullerenes as neuroprotective agents. Proc Natl Acad Sci USA 94:9434–9439
Jin H, Chen WQ, Tang XW, Chiang LY, Yang CY, Schloss JV, Wu JY (2000) Polyhydroxylated C(60), fullerenols, as glutamate receptor antagonists and neuroprotective agents. J Neurosci Res 62:600–607
Yang X, Chen Z, Meng X, Li B, Tan X (2007) Inhibition of DNA restrictive endonucleases and Taq DNA polymerase by trimalonic acid C60. Chin Sci Bull 52:1802–1806
Iwata N, Mukai T, Yamakoshi Y, Hara S, Yanase T, Shoji M, Endo T, Miyata N (1998) Effects of C60, a fullerene, on the activities of glutathione S-transferase and glutathione-related enzymes in rodent and human livers. Fullerenes Nanotubes Carbon Nanostruct 6:213–226
Lai YL, Chiang LY (1997) Water-soluble fullerene derivatives attenuate exsanguination-induced bronchoconstriction of guinea-pigs. J Auton Pharmacol 17:229–235
Nielsen GD, Roursgaard M, Jensen KA, Poulsen SS, Larsen ST (2008) In vivo biology and toxicology of fullerenes and their derivatives. Basic Clin Pharmacol Toxicol 103:197–208
Yang X, Meng X, Li B, Chen Z, Zhao D, Tan X, Yu Q (2008) Inhibition of in vitro amplification of targeted DNA fragment and activity of exonuclease I by a fullerene-oligonucleotide conjugate. Biologicals 36:223–226
Wang Z, Zhao J, Li F, Gao D, Xing B (2009) Adsorption and inhibition of acetylcholinesterase by different nanoparticles. Chemosphere 77:67–73
Innocenti A, Durdagi S, Doostdar N, Strom TA, Barron AR, Supuran CT (2010) Nanoscale enzyme inhibitors: fullerenes inhibit carbonic anhydrase by occluding the active site entrance. Bioorg Med Chem 18:2822–2828
Bosi S, Da Ros T, Castellano S, Banfi E, Prato M (2000) Antimycobacterial activity of ionic fullerene derivatives. Bioorg Med Chem Lett 10:1043–1045
Klumpp C, Lacerda L, Chaloin O, Da Ros T, Kostarelos K, Prato M, Bianco A (2007) Multifunctionalised cationic fullerene adducts for gene transfer: design, synthesis and DNA complexation. Chem Commun (Camb) 36:3762–3764
Bosi S, Da Ros T, Spallato G, Prato M (2003) Fullerene derivatives: an attractive tool for biological applications. Eur J Med Chem 38:913–923
Johnston HJ, Hutchison GR, Frans M, Christensen FM, Aschberger K, Stone V (2010) The biological mechanisms and physicochemical characteristics responsible for driving fullerene toxicity. Toxicol Sci 114:162–182
Trpkovic A, Todorovic-Markovic B, Kleut D, Misirkic M, Janjetovic K, Vucicevic L, Pantovic A, Jovanovic S, Dramicanin M, Markovic Z, Trajkovic V (2010) Oxidative stress-mediated hemolytic activity of solvent exchange-prepared fullerene (C60) nanoparticles. Nanotechnology 21:375102
Bosi S, Feruglio L, Da Ros T, Spalluto G, Gregoretti B, Terdoslavich M, Decorti G, Passamonti S, Moro S, Prato M (2004) Haemolytic effects of water-soluble fullerene derivatives. J Med Chem 47:6711–6715
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Tramer, F., Da Ros, T., Passamonti, S. (2012). Screening of Fullerene Toxicity by Hemolysis Assay. In: Reineke, J. (eds) Nanotoxicity. Methods in Molecular Biology, vol 926. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-002-1_15
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DOI: https://doi.org/10.1007/978-1-62703-002-1_15
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