Abstract
A finely ground fibrillated cellulose was fractionated into separate size fractions. The characteristics of the smallest size fractions were studied, and the toxicity to humans was tested as part of a safety assessment. Morphological studies performed with state-of-the-art methods, such as scanning electron microscopy and atomic force microscopy, showed that the fraction obtained consisted of long thin fibrils but also larger fibril agglomerates, and spherical particles were present. The finest fraction did not show any sub-lethal effects as assessed by RNA inhibition test in vitro, nor were there any indications of genotoxicity as tested by the Ames test in vitro. Systemic effects tested in vivo with the nematode were also absent. No cytotoxic effects were seen in the highest tolerated dose test in vitro, but some indication of cytotoxicity was observed in the total protein content test in vitro at the highest sample concentration. The significance of this toxicity test result should be addressed in relation to the other toxicity tests, in which no toxicity was observed, with special emphasis on the in vivo test. Given this, the overall toxicity analyses support the conclusion that nano-scale cellulose fibrils can be considered to be safe towards humans. However, the reason for the positive cytotoxicity test result and, in addition, the effect of the biocide used in sample preservation on the toxicity tests need to be clarified before generalizing these results and declaring nanocellulose to be unambiguously safe.
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Alexandrescu L, Syverud K, Gatti A, Chinga Carrasco G (2013) Cytotoxicity test of cellulose nanofibril-based structures. Cellulose 20:1765–1775. doi:10.1007/s10570-013-9948-9
Bhattacharya M, Malinen MM, Lauren P, Lou Y-R, Kuisma SW, Kanninen L, Lille M, Corlu A, Gu-Guen-Guillouzo C, Ikkala O, Laukkanen A, Urtti A, Yliperttula M (2012) Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture. J Control Release 164:291–298. doi:10.1016/j.jconrel.2012.06.039
ECHA European Chemicals Agency (2012) Guidance on information requirements and chemical safety assessment. Appendix R7-1 Recommendations for nanomaterials applicable to Chapter R7a Endpoint specific guidance, p 59
Forestcluster Ltd. (2011) Intelligent and resource-efficient production technologies. EffTech programme. Programme report 2008–2010, pp 119–120
Haapala AT, Laitinen O, Karinkanta P, Liimatainen H, Niinimäki J (2013) Optical characterization of size, shape and fibrillarity from microfibrillar and microcrystalline cellulose, and fine ground wood powder fractions. Appita J 66:331–339
Hasegawa K, Miwa S, Tsutsumiuchi K, Taniguchi H, Miwa J (2004) Extremely low dose of acrylamide decreases lifespan in Caenorhabditis elegans. Toxicol Lett 152:183–189
Hua K, Carlsson DO, Ålander E, Linström T, Stromme M, Mihranyan A, Ferraz N (2014) Translational study between structure and biological response of nanocellulose from wood and green algae. RSC Adv 4:2892–2903. doi:10.1039/c3ra45553j
Jeong SI, Lee SE, Yang H, Jin Y-H, Park C-S, Park YS (2010) Toxicologic evaluation of bacterial synthesized cellulose in endothelial cells and animals. Mol Cell Toxicol 6:373–380. doi:10.1007/s13273-010-0049-7
Kaletta T and Hengartner MO (2006) Finding function in novel targets: C. elegans as a model organism. Nat Rev Drug Discov. AOP published online 21 Apr 2006. doi:10.1038/nrd2031
Kangas H, Lahtinen P, Sneck A, Saariaho A-M, Laitinen O, Hellén E (2014) Characterization of fibrillated celluloses. A short review and evaluation of characteristics with a combination of methods. Nord Pulp Paper Res J 29:129–143
Khandoudi N, Porte P, Chtourou S, Nesslany F, Marzin D, Le Curieux F (2009) The presence of arginine may be a source of false positive results in the Ames test. Mutat Res 679:65–71. doi:10.1016/j.mrgentox.2009.03.010
Laitinen O, Niinimäki J (2014) Fractional study of the microfibrillated cellulose. Tappi J 13:49–55
Laitinen O, Kemppainen K, Stoor T, Niinimäki J (2011) Fractionation of pulp and paper particles selectively by size. BioResources 6:672–685
Landsiedel R, Kapp MD, Schulz M, Wiench K, Oesch F (2009) Genotoxicity investigations on nanomaterials: methods, preparation and characterization of test material, potential artifacts and limitations—many questions, some answers. Mutat Res 681:241–258. doi:10.1016/j.mrrev.2008.10.002
Moreira S, Silva NB, Almeida-Lima J, Rocha HAO, Medeiros SRB, Alves C, Cama FM (2009) BC nanofibers: in vitro study of genotoxicity and cell proliferation. Toxicol Lett 189:235–241
O’Connor B (2009) Ensuring safety of manufactured nanocrystalline cellulose. In: OECD conference, July 15–17, 2009, Paris
O’Connor B (2011) Ensuring safety of manufactured nanocrystalline cellulose. A risk assessment under Canada’s new substances notification regulations. In: 2011 TAPPI international conference on nanotechnology for renewable materials. June 6–8, 2011, Arlington, VA, USA
O’Connor B (2012) NCC: Environmental health and safety update. In: 2012 TAPPI international conference on nanotechnology for renewable materials. June 5–7, 2012, Montreal, Canada
Pertile RAN, Moreira S, da Costa RMG, Correia A, Guardao L, Gartner F, Vilanova M, Gama M (2012) Bacterial cellulose: long-term biocompatibility studies. J Biomater Sci 23:1339–1354
Pitkänen M, Sneck A, Hentze H-P, Sievänen J, Hiltunen J, Hellén E, Honkalampi U, von Wright A (2010) Nanofibrillar cellulose—Assessment of cytotoxic and genotoxic properties in vitro. In: 2010 Tappi International conference on nanotechnology for the forest products industry. Sept 27–29, 2010, Espoo, Finland
Pitkänen M, Kangas H, Vartiainen J (2014) Toxicity and health issues. In: Chapter 12 in Handbook of Green Materials; Processing Technologies, Properties and Applications, vol 1. Bionanomaterials: separation processes, characterization and properties. World Scientific, pp 181–205
Rouhiainen J, Tsitko I, Vippola M, Koivisto J (2010) Literature study on risks and risk assessment methods related to nanobased products and the recommended methodology for risk of nanofibrillar cellulose products. In: A public report of Scale-up Nanoparticles in Modern Papermaking-SUNPAP, FP7, Theme 4, NMP—Nanosciences, Nanotechnologies, Materials and New Production Technologies, p 53
Rouhiainen J, Väänänen V, Tsitko I, Kautto J (2012) Risk assessment of nanofibrillated cellulose in occupational settings. In: SUNPAP Final conference, June 19–20, 2012, Milan, Italy
Tanaka A, Seppänen V, Houni J, Sneck A, Pirkonen P (2012) Nanocellulose characterisation with mechanical fractionation. Nord Pulp Paper Res J 27:689–694
Vartiainen S (2007) Caenorhabditis elegans as a model for human synucleopathies. Doctoral Thesis, Kuopio University, Finland, 107 p
Vartiainen J, Vikman M (2013) Health and safety aspects of CNF. In: Postek MT, Moon RJ, Rudie AW, Bilodeau MA (eds) Production and Applications of Cellulose Nanonaterials. TAPPI PRESS, GA, USA
Vartiainen J, Pöhler T, Sirola K, Pylkkänen L, Alenius H, Hokkinen J, Tapper U, Lahtinen P, Kapanen A, Putkisto K, Hiekkataipale P, Eronen P, Ruokolainen J, Laukkanen A (2011) Health and environmental safety aspects of friction grinding and spray drying of microfibrillated cellulose. Cellulose 18:775–786. doi:10.1007/s10570-011-9501-7
Wang QQ, Zhung JY, Gleiner R, Kuster TA, Baxa U, McNeil SE (2012) Morphological development of cellulose fibrils of a bleached eucalyptus pulp by mechanical fibrillation. Cellulose 19:1631–1643. doi:10.1007/s10570-012-9745-x
Acknowledgments
The research leading to these results was performed as part of the Efficient Networking towards Novel Products and Processes (EffNet) research programme of the Finnish Bioeconomy Cluster (FIBIC). The EffNet programme has received funding from Tekes—the Finnish Funding Agency for Technology and Innovation and from Forestcluster Ltd. The authors would like to thank Ulla Honkalampi-Hämäläinen, Biosafe Special Laboratory Services Oy Ltd, and Professor Atte von Wright, University of Eastern Finland, for performing the toxicity tests and their help in interpretation of the results.
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Pitkänen, M., Kangas, H., Laitinen, O. et al. Characteristics and safety of nano-sized cellulose fibrils. Cellulose 21, 3871–3886 (2014). https://doi.org/10.1007/s10570-014-0397-x
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DOI: https://doi.org/10.1007/s10570-014-0397-x