Biocompatibility of Carbon Nanoparticles in HeLa Cells is Dictated by Synthesis and Sterilization Procedures
Due to their unique chemical and physical properties, carbon-based NanoMaterials (C-NMs) are largely exploited in biomedicine, i.e., cell and tissue imaging, drug delivery and tissue engineering scaffold, even if reports regarding their toxicity are still conflicting. In fact, biological effects strictly depend on the dynamic physicochemical characteristics of C-NMs, which in turn are strongly influenced by the procedures of their synthesis, and nanometrological techniques, e.g., Electron Microscopy (EM)-based analysis, are becoming the main tool for researchers to characterize nanoproducts. The aim of the present work is the study of the influence of synthesis and sterilization protocols on the size, shape, stability and biocompatibility of carbon NanoParticles (C-NPs). C-NPs were synthesized by using graphite as bulk material through an electrochemical method applying a constant voltage of 30 V and different times of synthesis. The C-NPs solution was sterilized by adopting different sterilization protocols during and/or after the synthesis. Size, shape and stability were studied by TEM and spectroscopy, while biocompatibility was tested in HeLa cells. Synthesis and sterilization procedures did not influence size, shape and stability of C-NPs, but interfered with C-NPs biocompatibility. In fact, irrespective of time of electrolysis process, the NPs show spherical shape with an average diameter of 7 nm. UV-visible spectra show typical peak of carbonaceous materials that falls at 236 nm without aggregation and sedimentation. However, when NPs obtained at 90 min of synthesis were twice autoclaved the peak shifted to 257 nm. HeLa cells were incubated with different C-NPs solutions administered at different concentrations, ranging from 8×105 to 1.6×107 C-NPs/cell, for different times (4, 24 and 48h). Cell viability was C-NPs concentration- and time of culture-dependent; interestingly, also the time of electrolysis process used during particles synthesis and procedures adopted to sterilize C-NPs solutions largely influenced cells response.
Biocompatibility of Carbon Nanoparticles in HeLa Cells is Dictated by Synthesis and Sterilization Procedures, Nanoscience and Nanometrology.
Vol. 2, No. 1,
2016, pp. 1-7.
A. Krueger, Carbon materials and nanotechnology. Wiley-VCH, Weinheim, 2010.
C. Berger, Z. M. Song, X. B. Li, X. S. Wu, N. Brown, C. Naud,D. Mayo, T. B. Li, J. Hass, A. N. Marchenkov, E. H. Conrad, P. N. First, and W. A. de Heer, "Electronic confinement and coherence in patterned epitaxial graphene," Science, vol. 312, pp. 1191-1196, 2006.
S. Gilje, H. Song, M. Wang, K. L. Wang, and R. B. Kaner, "A chemical route to graphene for device applications," Nano Lett., vol. 7, pp. 3394-3398, 2007.
Y. G. Guo, Y. S. Hu, and J. Maier, "Synthesis of hierarchically mesoporous anatase spheres and their application in lithium batteries," Chem. Commun., vol. 26, pp. 2783-2785, 2006.
Z. Y. Yuan and B. L. Su, "Insights into hierarchically meso-macroporous structured materials," J. Mater. Chem., vol. 16, pp. 663-677, 2006.
L. Quercia, F. Loffredo, B. Alfano, V. La Ferrara, and G. Di Francia, "Fabrication and characterization of carbon nanoparticles for polymer based vapor sensors," Sens. Actuators, B, vol. 100, pp. 22-28, 2004.
Y. Li, X. Fan, J. Qi, J. Ji, S. Wang, G. Zhang, and F. Zhang, "Palladium nanoparticle-graphene hybrids as active catalysts for the Suzuki reaction," Nano Res., vol. 3, pp. 429-437, 2010.
J. Lee, S. Yoon, T. Hyeon, S. M. Oh, and K. B. Kim, “Synthesis of a new mesoporous carbon and its application to electrochemical double-layer capacitors,” Chem. Commun., vol. 21, pp. 2177-2178, 1999.
H. Yang, Q. Shi, X. Liu, S. Xie, D. Jiang, F. Zhang, C. Yu, B. Tu, and D. Zhao, "Synthesis of ordered mesoporous carbon monoliths with bicontinuous cubic pore structure of Ia3d symmetry," Chem. Commun., vol. 23, pp. 2842-2843, 2002.
M. D. Stoller, S. Park, Y. Zhu, J. An, and R. S. Ruoff, "Graphene-based ultracapacitors," Nano Lett., vol. 8, pp. 3498-3502, 2008.
B. S. Harrison and A. Atala, “Carbon nanotube applications for tissue engineering,” Biomaterials, vol. 28, pp. 344-353, 2007.
L. Cao, X. Wang, M. J. Meziani, F. Lu, H. Wang, P. G. Luo, Y. Lin, B. A. Harruff, L. M. Veca, D. Murray, S. Y. Xie, and Y. P. Sun, "Carbon dots for multiphoton bioimaging," J. Am. Chem. Soc., vol. 129, pp. 11318-11319, 2007.
Y. P. Sun, B. Zhou, Y. Lin, W. Wang, K. A. Fernando, S. P. Pathak,M. J. Meziani, B. A. Harruff, X. Wang, H. Wang, P. G. Luo,H. Yang, M. E. Kose, B. Chen, L. M. Veca, and S. Y. Xie, "Quantum-sized carbon dots for bright and colorful photoluminescence," J. Am. Chem. Soc., vol. 128, pp. 7756-7757, 2006.
T. W. Kim, P. W. Chung, I. I. Slowing, M. Tsunoda, E. S. Yeung, and V. S. Y. Lin, "Structurally ordered mesoporous carbon nanoparticles as transmembrane delivery vehicle in human cancer cells," Nano Lett., vol. 8, pp. 3724-3727, 2008.
Z. Liu, J. T. Robinson, X. Sun, and H. Dai, "PEGylated nanographene oxide for delivery of water-insoluble cancer drugs," J. Am. Chem. Soc., vol. 130, pp. 10876-10877, 2008.
H. Liu, T. Ye, and C. Mao, "Fluorescent carbon nanoparticles derived from candle soot," Angew. Chem., Int. Ed., vol. 46, pp. 6473- 6475, 2007.
A. Galvez, N. Herlin-Boimeb, C. Reynaudb, C. Clinarda, and J. N. Rouzaud, "Carbon nanoparticles from laser pyrolysis," Carbon, vol. 40, pp. 2775-2789, 2002.
Y. Yan, H. Yang, F. Zhang, B. Tu, and D. Zhao, "Low-temperature solution synthesis of carbon nanoparticles, onions and nanopores by the assembly of aromatic molecules," Carbon, vol. 45, pp. 2209-2216, 2007.
Q. L. Zhao, Z. L. Zhang, B. H. Huang, J. Peng, M. Zhang, and D. W. Pang, "Facile preparation of low cytotoxicity fluorescent carbon nanocrystals by electrooxidation of graphite," Chem. Commun., vol. 41, pp. 5116-5118, 2008.
H. Zhu, X. Wang, Y. Li, Z. Wang, F. Yang, and X. Yang, "Microwave synthesis of fluorescent carbon nanoparticles with electrochemiluminescence properties," Chem. Commun., vol. 34, pp. 5118-5120, 2009.
S. J. Yu, M. W. Kang, H. C. Chang, K. M. Chen, and Y. C. Yu, "Bright fluorescent nanodiamonds: no photobleaching and low cytotoxicity," J. Am. Chem. Soc., vol. 127, pp. 17604-17605, 2005.
R. Selvi, D. Jagadeesan, B. S. Suma, G. Nagashankar, M. Arif, K. Balasubramanyam, M. Eswaramoorthy, and T. K. Kundu, "Intrinsically fluorescent carbon nanospheres as a nuclear targeting vector: delivery of membrane-impermeable molecule to modulate gene expression in vivo," Nano Lett., vol. 8, pp. 3182-3188, 2008.
V. N. Mochalin and Y. Gogotsi, "Wet chemistry route to hydrophobic blue fluorescent nanodiamond," J. Am. Chem. Soc., vol. 131, pp. 4594-4595, 2009.
Z. Wang, F. Li, and A. Stein, "Direct synthesis of shaped carbon nanoparticles with ordered cubic mesostructure," Nano Lett., vol. 7, pp. 3223-3226, 2007.
Tang, K. Qi, K. L. Wooley, K. Matyjaszewski, and T. Kowalewski, "Well-Defined Carbon Nanoparticles Prepared from Water-Soluble Shell Cross-linked Micelles that Contain Polyacrylonitrile Cores," Angewandte Chemie, vol. 116, pp. 2843-2847, 2004.
L. Dini, E. Panzarini, S. Mariano, D. Passeri, M. Reggente, M. Rossi, and C. Vergallo, “Microscopies at the nanoscale for nano-scale drug delivery systems,” Curr. Drug Targets, 2015, in press.
K. Donaldson, L. Tran, L. A. Jimenez, R. Duffin, D. E. Newby, N. Mills, W. MacNee, and V. Stone, "Combustion-derived nanoparticles: a review of their toxicology following inhalation exposure," Part. Fibre Toxicol., vol. 2, p. 10, 2005.
D. Manno, E. Carata, B.A. Tenuzzo, E. Panzarini, A. Buccolieri, E. Filippo, M. Rossi, A. Serra, and L. Dini, “High ordered biomineralization induced by carbon nanoparticles in the sea urchin Paracentrotus lividus,” Nanotechnology, vol. 23, p. 495104, 2012.
E. Carata, B. Anna Tenuzzo, F. Arnò, A. Buccolieri, A. Serra, D. Manno, and L. Dini, “Stress response induced by carbon nanoparticles in Paracentrotus lividus,” Int. J. Mol. Cell. Med., vol. 1, pp. 30-38, 2012.
D. Manno, A. Serra, A. Buccolieri, E. Panzarini, E. Carata, B. Tenuzzo, D. Izzo, C. Vergallo, M. Rossi, and L. Dini, “Silver and carbon nanoparticles toxicity in sea urchin Paracentrotus lividus embryos,” BioNanoMat., vol. 14, pp. 229-238, 2013.
A. Bianco, K. Kostarelos, C. D. Partidos, and M. Prato, “Biomedical applications of functionalised carbon nanotubes,” Chem Commun (Camb) vol. 5, pp. 571-577, 2005.
S. R. Shin, H. Bae, J. M. Cha, J.Y. Mun, Y.C. Chen, H. Tekin, H. Shin, S. Farshchi, M. R. Dokmeci, S. Tang, and A. Khademhosseini, “Carbon nanotube reinforced hybrid microgels as scaffold materials for cell encapsulation,” ACS Nano., vol. 6, pp. 362-372, 2012.
C. Cha, S.R. Shin, N. Annabi, M.R. Dokmeci, and A. Khademhosseini, “Carbon-based nanomaterials: multifunctional materials for biomedical engineering,” ACS Nano, vol. 7, pp. 2891-2897, 2013.
S.T. Yang, J. Luo, Q. Zhou, and H. Wang, “Pharmacokinetics, metabolism and toxicity of carbon nanotubes for biomedical purposes,” Theranostics, vol. 2, pp. 271-282, 2012.
C.W. Lam, J.T. James, R. Mc Cluskey, S. Arepalli, and R.L. Hunter, “A review of carbon nanotube toxicity and assessment of potential occupational and environmental health risks,” Crit. Rev. Toxicol., vol. 36, pp. 189-217, 2006.
C.P. Firme and P.R. Bandaru, “Toxicity issues in the application of carbon nanotubes to biological systems,” Nanomedicine, vol. 6, pp. 245-56, 2010.