[KSCI] Korea Science Citation Index Service

http://dx.doi.org/10.5487/TR.2019.35.1.045

Inhalation of Bacterial Cellulose Nanofibrils Triggers an Inflammatory Response and Changes Lung Tissue Morphology of Mice

Silva-Carvalho, Ricardo (CEB - Centre of Biological Engineering, University of Minho)
Silva, Joao P. (UCIBIO, REQUIMTE - Laboratory of Toxicology, Department of Biological Sciences, Faculty of Pharmacy, University of Porto)
Ferreirinha, Pedro (ICBAS - Biomedical Sciences Institute Abel Salazar, University of Porto)
Leitao, Alexandre F. (CEB - Centre of Biological Engineering, University of Minho)
Andrade, Fabia K. (Embrapa Tropical Agroindustry)
da Costa, Rui M. Gil (LEPAE - Laboratory for Process, Environmental and Energy Engineering, Chemical Engineering Department, Faculty of Engineering, University of Porto)
Cristelo, Cecilia (CEB - Centre of Biological Engineering, University of Minho)
Rosa, Morsyleide F. (Embrapa Tropical Agroindustry)
Vilanova, Manuel (ICBAS - Biomedical Sciences Institute Abel Salazar, University of Porto)
Gama, F. Miguel (CEB - Centre of Biological Engineering, University of Minho)

Publication Information

Toxicological Research / v.35, no.1, 2019 , pp. 45-63 More about this Journal

Abstract

In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages ( $BMM{\Phi}$ ) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and $10{\mu}g$ of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline $Avicel-plus^{(R)}$ CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. $Avicel-plus^{(R)}$ CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.

Keywords

Bacterial cellulose; Airborne nanofibers; Lung toxicity; Inflammation;

Citations & Related Records

Reference

1	Fröhlich, E., Mercuri, A., Wu, S. and Salar-Behzadi, S. (2016) Measurements of deposition, lung surface area and lung fluid for simulation of inhaled compounds. Front. Pharmacol., 7, 181.
2	Task Group of the International Commission on Radiological Protection. (1994) ICRP Human Respiratory Tract Model for Radiological Protection, Ann. ICRP.
3	Bates, D.V., Fish, B.R., Hatch, T.F., Mercer, T.T. and Morrow, P.E. (1966) Deposition and retention models for internal dosimetry of the human respiratory tract. Task group on lung dynamics. Health Phys., 12, 173-207.
4	Beppu, H., Ichinose, F., Kawai, N., Jones, R.C., Yu, P.B., Zapol, W.M., Miyazono, K., Li, E. and Bloch, K.D. (2004) BMPR-II heterozygous mice have mild pulmonary hypertension and an impaired pulmonary vascular remodeling response to prolonged hypoxia. Am. J. Physiol. Lung Cell Mol. Physiol., 287, L1241-L1247. DOI
5	Tátrai, E., Brozik, M., Adamis, Z., Meretey, K. and Ungvary, G. (1996) In vivo pulmonary toxicity of cellulose in rats. J. Appl. Toxicol., 16, 129-135. DOI
6	Muhle, H., Ernst, H. and Bellmann, B. (1997) Investigation of the durability of cellulose fibres in rat lungs. Ann. Occup. Hyg., 41, 184-188. DOI
7	Jeong, S.I., Lee, S.E., Yang, H., Jin, Y.-H., Park, C.-S. and Park, Y.S. (2010) Toxicologic evaluation of bacterial synthesized cellulose in endothelial cells and animals. Mol. Cell. Toxicol., 6, 370-377. DOI
8	Vasconcelos, N.F., Feitosa, J.P., da Gama, F.M., Morais, J.P., Andrade, F.K., de Souza Filho, M.S. and Rosa, M.F. (2017) Bacterial cellulose nanocrystals produced under different hydrolysis conditions: Properties and morphological features. Carbohydr. Polym., 155, 425-431. DOI
9	Martínez-Sanz, M., Olsson, R.T., Lopez-Rubio, A. and Lagaron, J.M. (2012) Development of bacterial cellulose nanowhiskers reinforced EVOH composites by electrospinning. J. Appl. Polym. Sci., 124, 1398-1408. DOI
10	Svensson, A., Nicklasson, E., Harrah, T., Panilaitis, B., Kaplan, D.L., Brittberg, M. and Gatenholm, P. (2005) Bacterial cellulose as a potential scaffold for tissue engineering of cartilage. Biomaterials, 26, 419-431. DOI
11	Leitão, A., Silva, J., Dourado, F. and Gama, M. (2013) Production and characterization of a new bacterial cellulose/ poly(vinyl alcohol) nanocomposite. Materials, 6, 1956-1966. DOI
12	Ummartyotin, S. and Manuspiya, H. (2015) A critical review on cellulose: from fundamental to an approach on sensor technology. Renew. Sust. Energ. Rev., 41, 402-412. DOI
13	Solway, D.R., Consalter, M. and Levinson, D.J. (2010) Microbial cellulose wound dressing in the treatment of skin tears in the frail elderly. Wounds, 22, 17-19.
14	Fontana, J.D., de Souza, A.M., Fontana, C.K., Torriani, I.L., Moreschi, J.C., Gallotti, B.J., de Souza, S.J., Narcisco, G.P., Bichara, J.A. and Farah, L.F. (1990) Acetobacter cellulose pellicle as a temporary skin substitute. Appl. Biochem. Biotechnol., 24-25, 253-264.
15	Andrade, F.K., Costa, R., Domingues, L., Soares, R. and Gama, M. (2010) Improving bacterial cellulose for blood vessel replacement: Functionalization with a chimeric protein containing a cellulose-binding module and an adhesion peptide. Acta Biomater., 6, 4034-4041. DOI
16	Backdahl, H., Helenius, G., Bodin, A., Nannmark, U., Johansson, B.R., Risberg, B. and Gatenholm, P. (2006) Mechanical properties of bacterial cellulose and interactions with smooth muscle cells. Biomaterials, 27, 2141-2149. DOI
17	Moon, R.J., Martini, A., Nairn, J., Simonsen, J. and Youngblood, J. (2011) Cellulose nanomaterials review: structure, properties and nanocomposites. Chem. Soc. Rev., 40, 3941-3994. DOI
18	Stefaniak, A.B., Seehra, M.S., Fix, N.R. and Leonard, S.S. (2014) Lung biodurability and free radical production of cellulose nanomaterials. Inhal. Toxicol., 26, 733-749. DOI
19	Bäckdahl, H., Risberg, B. and Gatenholm, P. (2011) Observations on bacterial cellulose tube formation for application as vascular graft. Mater. Sci. Eng. C., 31, 14-21. DOI
20	Leitao, A.F., Gupta, S., Silva, J.P., Reviakine, I. and Gama, M. (2013) Hemocompatibility study of a bacterial cellulose/polyvinyl alcohol nanocomposite. Colloids Surf. B Biointerfaces, 111, 493-502. DOI
21	Andrade, F.K., Pertile, R.A.N., Dourado, F. and Gama, M. (2010) Bacterial cellulose: properties, production and applications in Cellulose: Structure and Properties, Derivatives and Industrial Uses (Lejeune, A. and Deprez, T. Eds). Nova Science Publishers, Inc., pp. 427-458.
22	Klemm, D., Schumann, D., Udhardt, U. and Marsch, S. (2001) Bacterial synthesized cellulose - artificial blood vessels for microsurgery. Prog. Polym. Sci., 26, 1561-1603. DOI
23	Fu, L.N., Wang, W., Yu, L.J., Zhang, S.M. and Yang, G. (2009) Fabrication of novel cellulose/chitosan artificial skin composite. Mater. Sci. Forum, 610-613, 1034-1038. DOI
24	Millon, L.E., Guhados, G. and Wan, W. (2008) Anisotropic polyvinyl alcohol-Bacterial cellulose nanocomposite for biomedical applications. J. Biomed. Mater. Res. Part B Appl. Biomater., 86, 444-452. DOI
25	Martinez-Sanz, M., Lopez-Rubio, A. and Lagaron, J.M. (2012) Optimization of the dispersion of unmodified bacterial cellulose nanowhiskers into polylactide via melt compounding to significantly enhance barrier and mechanical properties. Biomacromolecules, 13, 3887-3899. DOI
26	Pitkanen, M., Kangas, H., Laitinen, O., Sneck, A., Lahtinen, P., Peresin, M.S. and Niinimaki, J. (2014) Characteristics and safety of nano-sized cellulose fibrils. Cellulose, 21, 3871-3886. DOI
27	Zhou, C. and Wu, Q. (2012) Recent development in applications of cellulose nanocrystals for advanced polymer-based nanocomposites by novel fabrication strategies in Nanocrystals: Synthesis, Characterization and Applications (Neralla, S. Ed). IntechOpen.
28	Charreau, H., Foresti, M.L. and Vazquez, A. (2013) Nanocellulose patents trends: a comprehensive review on patents on cellulose nanocrystals, microfibrillated and bacterial cellulose. Recent Pat. Nanotechnol., 7, 56-80. DOI
29	Li, Y. and Ragauskas, A.J. (2011) Cellulose nano whiskers as a reinforcing filler in polyurethanes in Avances in Diverse Industrial Applications Of Nanocomposites (Reddy, B. Ed.). IntechOpen, pp. 19-36.
30	Dugan, J.M., Gough, J.E. and Eichhorn, S.J. (2013) Bacterial cellulose scaffolds and cellulose nanowhiskers for tissue engineering. Nanomedicine (Lond.), 8, 287-298. DOI
31	Martínez-Sanz, M., Lopez-Rubio, A. and Lagaron, J.M. (2011) Optimization of the nanofabrication by acid hydrolysis of bacterial cellulose nanowhiskers. Carbohydr. Polym., 85, 228-236. DOI
32	Nechyporchuk, O., Belgacem, M.N. and Bras, J. (2016) Production of cellulose nanofibrils: a review of recent advances. Ind. Crops Prod., 93, 2-25. DOI
33	Cullen, R., Miller, B., Jones, A. and Davis, J. (2002) Toxicity of cellulose fibres. Ann. Occup. Hyg., 46, 81-84. DOI
34	Future Markets Inc. (2012) Nanocellulose: A Technology and Market Study, pp. 35.
35	Hull, M. and Bowman, D. (2014) Nanotechnology Environmental Health and Safety (2nd edition), Elsevier.
36	Cullen, R.T., Miller, B.G., Jones, A.D. and Davis, J.M.G. (2002) Toxicity of cellulose fibres. Ann. Occup. Hyg., 46, 81-84. DOI
37	Kovacs, T., Naish, V., O'Connor, B., Blaise, C., Gagne, F., Hall, L., Trudeau, V. and Martel, P. (2010) An ecotoxicological characterization of nanocrystalline cellulose (NCC). Nanotoxicology, 4, 255-270. DOI
38	Shi, Q., Li, Y., Sun, J., Zhang, H., Chen, L., Chen, B., Yang, H. and Wang, Z. (2012) The osteogenesis of bacterial cellulose scaffold loaded with bone morphogenetic protein-2. Biomaterials, 33, 6644-6649. DOI
39	Zaborowska, M., Bodin, A., Bäckdahl, H., Popp, J., Goldstein, A. and Gatenholm, P. (2010) Microporous bacterial cellulose as a potential scaffold for bone regeneration. Acta Biomater., 6, 2540-2547. DOI
40	Cullen, R.T., Searl, A., Miller, B.G., Davis, J.M. and Jones, A.D. (2000) Pulmonary and intraperitoneal inflammation induced by cellulose fibres. J. Appl. Toxicol., 20, 49-60. DOI
41	Catalan, J., Rydman, E., Aimonen, K., Hannukainen, K.S., Suhonen, S., Vanhala, E., Moreno, C., Meyer, V., Perez, D.D., Sneck, A., Forsstrom, U., Hojgaard, C., Willemoes, M., Winther, J.R., Vogel, U., Wolff, H., Alenius, H., Savolainen, K.M. and Norppa, H. (2016) Genotoxic and inflammatory effects of nanofibrillated cellulose in murine lungs. Mutagenesis, 32, 23-32. DOI
42	Yanamala, N., Farcas, M.T., Hatfield, M.K., Kisin, E.R., Kagan, V.E., Geraci, C.L. and Shvedova, A.A. (2014) In vivo evaluation of the pulmonary toxicity of cellulose nanocrystals: a renewable and sustainable nanomaterial of the future. ACS Sustain. Chem. Eng., 2, 1691-1698. DOI
43	Schramm, M. and Hestrin, S. (1954) Synthesis of cellulose by Acetobacter xylinum. I. Micromethod for the determination of celluloses. Biochem. J., 56, 163-166. DOI
44	Mullane, K.M., Kraemer, R. and Smith, B. (1985) Myeloperoxidase activity as a quantitative assessment of neutrophil infiltration into ischemic myocardium. J. Pharmacol. Methods, 14, 157-167. DOI
45	Delgado, A.V., Gonzalez-Caballero, F., Hunter, R.J., Koopal, L.K. and Lyklema, J. (2007) Measurement and interpretation of electrokinetic phenomena. J. Colloid. Interface Sci., 309, 194-224. DOI
46	Segal, L., Creely, J.J., Martin, A.E. and Conrad, C.M. (1959) An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer. Text. Res. J., 29, 786-794. DOI
47	Torrado, E., Fraga, A.G., Logarinho, E., Martins, T.G., Carmona, J.A., Gama, J.B., Carvalho, M.A., Proenca, F., Castro, A.G. and Pedrosa, J. (2010) IFN-gamma-dependent activation of macrophages during experimental infections by Mycobacterium ulcerans is impaired by the toxin mycolactone. J. Immunol., 184, 947-955. DOI
48	Renne, R., Brix, A., Harkema, J., Herbert, R., Kittel, B., Lewis, D., March, T., Nagano, K., Pino, M., Rittinghausen, S., Rosenbruch, M., Tellier, P. and Wohrmann, T. (2009) Proliferative and nonproliferative lesions of the rat and mouse respiratory tract. Toxicol. Pathol., 37, 5S-73S. DOI
49	Lee, K.-Y., Quero, F., Blaker, J.J., Hill, C.A.S., Eichhorn, S.J. and Bismarck, A. (2011) Surface only modification of bacterial cellulose nanofibres with organic acids. Cellulose, 18, 595-605. DOI
50	Mirhosseini, H., Tan, C.P., Hamid, N.S.A. and Yusof, S. (2008) Effect of Arabic gum, xanthan gum and orange oil contents on ${\zeta}$ -potential, conductivity, stability, size index and pH of orange beverage emulsion. Colloids Surf. A, 315, 47-56. DOI
51	U.S. Food and Drug Administration (1987) Guideline on Validation of the Limulus Amebocyte Lysate Test as an Endproduct Endotoxin Test for Human and Animal Parenteral Drugs, Biological Products and Medical Devices, US Department of Health and Human Services, Public Health Service, Food and Drug Administration.
52	Duarte, E.B., das Chagas, B.S., Andrade, F.K., Brígida, A.I.S., Borges, M.F., Muniz, C.R., Souza Filho, M.d.S.M., Morais, J.P.S., Feitosa, J.P.A. and Rosa, M.F. (2015) Production of hydroxyapatite-bacterial cellulose nanocomposites from agroindustrial wastes. Cellulose, 22, 3177-3187. DOI
53	Raetz, C.R. and Whitfield, C. (2002) Lipopolysaccharide endotoxins. Annu. Rev. Biochem., 71, 635-700. DOI
54	Cundell, A.M. (2014) Bacterial endotoxin requirements for dry powder inhalants and their excipients: are they critical quality attributes? PDA J. Pharm. Sci. Technol., 68, 386-393. DOI
55	Vartiainen, J., Pohler, T., Sirola, K., Pylkkanen, L., Alenius, H., Hokkinen, J., Tapper, U., Lahtinen, P., Kapanen, A., Putkisto, K., Hiekkataipale, P., Eronen, P., Ruokolainen, J. and Laukkanen, A. (2011) Health and environmental safety aspects of friction grinding and spray drying of microfibrillated cellulose. Cellulose, 18, 775-786. DOI
56	Lopes, V.R., Sanchez-Martinez, C., Stromme, M. and Ferraz, N. (2017) In vitro biological responses to nanofibrillated cellulose by human dermal, lung and immune cells: surface chemistry aspect. Part. Fibre Toxicol., 14, 1. DOI
57	Kim, G.-D., Yang, H., Park, H.R., Park, C.-S., Park, Y.S. and Lee, S.E. (2013) Evaluation of immunoreactivity of in vitro and in vivo models against bacterial synthesized cellulose to be used as a prosthetic biomaterial. BioChip J., 7, 201-209. DOI
58	Endes, C., Mueller, S., Kinnear, C., Vanhecke, D., Foster, E.J., Petri-Fink, A., Weder, C., Clift, M.J. and Rothen- Rutishauser, B. (2015) Fate of cellulose nanocrystal aerosols deposited on the lung cell surface in vitro. Biomacromolecules, 16, 1267-1275. DOI
59	Zhang, Z., Ortiz, O., Goyal, R. and Kohn, J. (2014) Chapter 23 - biodegradable polymers A2 - Lanza, Robert in Principles of Tissue Engineering (4th edition) (Langer, R. and Vacanti, J. Eds.). Academic Press, Boston, pp. 441-473.
60	Hasenberg, M., Stegemann-Koniszewski, S. and Gunzer, M. (2013) Cellular immune reactions in the lung. Immunol. Rev., 251, 189-214. DOI
61	Labiris, N.R. and Dolovich, M.B. (2003) Pulmonary drug delivery. Part I: physiological factors affecting therapeutic effectiveness of aerosolized medications. Br. J. Clin. Pharmacol., 56, 588-599. DOI
62	Mantecca, P., Sancini, G., Moschini, E., Farina, F., Gualtieri, M., Rohr, A., Miserocchi, G., Palestini, P. and Camatini, M. (2009) Lung toxicity induced by intratracheal instillation of size-fractionated tire particles. Toxicol. Lett., 189, 206-214. DOI
63	Meng, G., Zhao, J., Wang, H.M., Ding, R.G., Zhang, X.C., Huang, C.Q. and Ruan, J.X. (2010) Cell injuries of the blood-air barrier in acute lung injury caused by perfluoroisobutylene exposure. J. Occup. Health, 52, 48-57. DOI
64	Noel-Georis, I., Bernard, A., Falmagne, P. and Wattiez, R. (2001) Proteomics as the tool to search for lung disease markers in bronchoalveolar lavage. Dis. Markers, 17, 271-284. DOI
65	Henderson, R.F., Damon, E.G. and Henderson, T.R. (1978) Early damage indicators in the the lung I. Lactate dehydrogenase activity in the airways. Toxicol. Appl. Pharmacol., 44, 291-297. DOI
66	Swedin, L., Arrighi, R. and ersson-Willman, B., Murray, A., Chen, Y., Karlsson, M.C.I., Georén, S.K., Tkach, A.V., Shvedova, A.A., Fadeel, B., Barragan, A. and Scheynius, A. (2012) Pulmonary exposure to single-walled carbon nanotubes does not affect the early immune response against Toxoplasma gondii. Part. Fibre Toxicol., 9, 16. DOI
67	Clift, M.J., Foster, E.J., Vanhecke, D., Studer, D., Wick, P., Gehr, P., Rothen-Rutishauser, B. and Weder, C. (2011) Investigating the interaction of cellulose nanofibers derived from cotton with a sophisticated 3D human lung cell coculture. Biomacromolecules, 12, 3666-3673. DOI
68	Adamis, Z., Tatrai, E., Honma, K. and Ungvary, G. (1997) In vitro and in vivo assessment of the pulmonary toxicity of cellulose. J. Appl. Toxicol., 17, 137-141. DOI
69	Chaplin, D.D. (2010) Overview of the immune response. J. Allergy Clin. Immunol., 125, S3-23. DOI
70	Liimatainen, H., Haavisto, S., Haapala, A. and Niinimaki, J. (2009) Influence of adsorbed and dissolved carboxymethyl cellulose on fibre suspension dispersing, dewaterability, and fines retention. Bioresources, 4, 321-340.
71	Winter, H.T., Cerclier, C., Delorme, N., Bizot, H., Quemener, B. and Cathala, B. (2010) Improved colloidal stability of bacterial cellulose nanocrystal suspensions for the elaboration of spin-coated cellulose-based model surfaces. Biomacromolecules, 11, 3144-3151. DOI
72	Khaled, B. and Abdelbaki, B. (2012) Rheological and electrokinetic properties of carboxymethylcellulose-water dispersions in the presence of salts. Int. J. Phys. Sci., 7, 1790 - 1798.
73	He, M., Cho, B.-U., Lee, Y.K. and Won, J.M. (2016) Utilizing cellulose nanofibril as an eco-friendly flocculant for filler flocculation in papermaking. BioRes, 11, 10296-10313.
74	Sheykhnazari, S., Tabarsa, T., Ashori, A., Shakeri, A. and Golalipour, M. (2011) Bacterial synthesized cellulose nanofibers; Effects of growth times and culture mediums on the structural characteristics. Carbohydr. Polym., 86, 1187-1191. DOI
75	Gama, M., Gatenholm, P. and Klemm, D. (2012) Bacterial NanoCellulose: a Sophisticated Multifunctional Material., CRC Press, Boca Raton, FL.
76	Malyala, P. and Singh, M. (2008) Endotoxin limits in formulations for preclinical research. J. Pharm. Sci., 97, 2041-2044. DOI
77	Kose, R., Mitani, I., Kasai, W. and Kondo, T. (2011) "Nanocellulose" as a single nanofiber prepared from pellicle secreted by Gluconacetobacter xylinus using aqueous counter collision. Biomacromolecules, 12, 716-720. DOI
78	Saito, T., Nishiyama, Y., Putaux, J.L., Vignon, M. and Isogai, A. (2006) Homogeneous suspensions of individualized microfibrils from TEMPO-catalyzed oxidation of native cellulose. Biomacromolecules, 7, 1687-1691. DOI
79	Lavoine, N., Desloges, I., Dufresne, A. and Bras, J. (2012) Microfibrillated cellulose - its barrier properties and applications in cellulosic materials: a review. Carbohydr. Polym., 90, 735-764. DOI
80	U.S. Food and Drug Administration (2012) Guidance for Industry: Pyrogen and Endotoxins Testing: Questions and Answers, Silver Spring, MD, p. 11.
81	Martinez Avila, H., Schwarz, S., Feldmann, E.M., Mantas, A., von Bomhard, A., Gatenholm, P. and Rotter, N. (2014) Biocompatibility evaluation of densified bacterial nanocellulose hydrogel as an implant material for auricular cartilage regeneration. Appl. Microbiol. Biotechnol., 98, 7423-7435. DOI
82	Bodin, A., Bharadwaj, S., Wu, S., Gatenholm, P., Atala, A. and Zhang, Y. (2010) Tissue-engineered conduit using urinederived stem cells seeded bacterial cellulose polymer in urinary reconstruction and diversion. Biomaterials, 31, 8889- 8901. DOI
83	Leitão, A.F., Faria, M.A., Faustino, A.M., Moreira, R., Mela, P., Loureiro, L., Silva, I. and Gama, M. (2016) A novel small-caliber bacterial cellulose vascular prosthesis: production, characterization, and preliminary in vivo testing. Macromol. Biosci., 16, 139-150. DOI
84	Shvedova, A.A., Kisin, E.R., Yanamala, N., Farcas, M.T., Menas, A.L., Williams, A., Fournier, P.M., Reynolds, J.S., Gutkin, D.W., Star, A., Reiner, R.S., Halappanavar, S. and Kagan, V.E. (2016) Gender differences in murine pulmonary responses elicited by cellulose nanocrystals. Part. Fibre Toxicol., 13, 28. DOI
85	Moreira, S., Silva, N.B., Almeida-Lima, J., Rocha, H.A., Medeiros, S.R., Alves, C., Jr. and Gama, F.M. (2009) BC nanofibres: in vitro study of genotoxicity and cell proliferation. Toxicol. Lett., 189, 235-241. DOI
86	Occupational Safety and Health Administration (OSHA) of the United States Department of Labor. CFR Code of Federal Regulations, U.S. Government Printing Office, Office of the Federal Register, Washington, DC.
87	Yanamala, N., Hatfield, M.K., Farcas, M.T., Schwegler-Berry, D., Hummer, J.A., Shurin, M.R., Birch, M.E., Gutkin, D.W., Kisin, E., Kagan, V.E., Bugarski, A.D. and Shvedova, A.A. (2013) Biodiesel versus diesel exposure: enhanced pulmonary inflammation, oxidative stress, and differential morphological changes in the mouse lung. Toxicol. Appl. Pharmacol., 272, 373-383. DOI