Effect of MWCNT size, carboxylation, and purification on in vitro and in vivo toxicity, inflammation and lung pathology

Springer Science and Business Media LLC - 2013
Raymond F. Hamilton1, Zheqiong Wu2, Somenath Mitra2, Pamela Shaw1, Andrij Holian1
1Department of Biomedical and Pharmaceutical Sciences, Center for Environmental Health Sciences, University of Montana, Missoula, MT, 59812, USA
2Department of Chemistry and Environmental Science, New Jersey Institute of Technology, Newark, NJ 07102, USA

Tóm tắt

Abstract Background Several properties of multi-walled carbon nanotubes (MWCNT) have the potential to affect their bioactivity. This study examined the in vitro and in vivo outcomes of the influence of diameter, length, purification and carboxylation (in vitro testing only) of MWCNT. Methods Three original ‘as received’ MWCNT that varied in size (diameter and length) were purified and functionalized by carboxylation. The resulting MWCNT were characterized and examined for cytotoxicity and inflammasome activation in vitro using THP-1 cells and primary alveolar macrophages from C57BL/6 mice. Oropharyngeal aspiration administration was used to deliver original MWCNT and in vivo bioactivity and lung retention was examined at 1 and 7 days. Results Studies with THP-1 macrophages demonstrated that increased length or diameter corresponded with increased bioactivity as measured by inflammasome activation. Purification had little effect on the original MWCNT, and functionalization completely eliminated bioactivity. Similar results were obtained using alveolar macrophages isolated from C57BL/6 mice. The in vivo studies demonstrated that all three original MWCNT caused similar neutrophil influx at one day, but increasing length or diameter resulted in the lavaged cells to release more inflammatory cytokines (IL-6, TNF-α, and IL-1β) ex vivo. Seven-day histology revealed that, consistent with the in vitro results, increasing width or length of MWCNT caused more severe pathology with the longest MWCNT causing the most severe inflammation. In addition, the same two larger MWCNT were retained more in the lung at 7 days. Conclusions Taken together, the results indicated that in vitro and in vivo bioactivity of MWCNT increased with diameter and length. Purification had no significant modifying effect from the original MWCNT. Functionalization by carboxylation completely eliminated the bioactive potential of the MWCNT regardless of size in in vitro testing.

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Donaldson K, Aitken R, Tran L, Stone V, Duffin R, Forrest G, Alexander A: Carbon nanotubes: a review of their properties in relation to pulmonary toxicology and workplace safety. Toxicol Sci 2006,92(1):5–22.

Murphy FA, Poland CA, Duffin R, Al-Jamal KT, Ali-Boucetta H, Nunes A, Byrne F, Prina-Mello A, Volkov Y, Li S, et al.: Length-dependent retention of carbon nanotubes in the pleural space of mice initiates sustained inflammation and progressive fibrosis on the parietal pleura. Am J Pathol 2011,178(6):2587–2600.

Palomaki J, Valimaki E, Sund J, Vippola M, Clausen PA, Jensen KA, Savolainen K, Matikainen S, Alenius H: Long, needle-like carbon nanotubes and asbestos activate the NLRP3 inflammasome through a similar mechanism. ACS Nano 2011,5(9):6861–6870.

Fenoglio I, Aldieri E, Gazzano E, Cesano F, Colonna M, Scarano D, Mazzucco G, Attanasio A, Yakoub Y, Lison D, et al.: Thickness of multiwalled carbon nanotubes affects their lung toxicity. Chem Res Toxicol 2012,25(1):74–82.

Qu GB, Bai YH, Zhang Y, Jia Q, Zhang WD, Yan B: The effect of multiwalled carbon nanotube agglomeration on their accumulation in and damage to organs in mice. Carbon N Y 2009,47(8):2060–2069.

Hamilton RF, Girtsman TA, Xiang C, Wu N, Holian A: Nickel contamination on MWCNT is related to particle bioactivity but not toxicity in the THP-1 transformed macrophage model. IJBNN 2013,3(1/2):107–126.

Liu X, Guo L, Morris D, Kane AB, Hurt RH: Targeted Removal of Bioavailable Metal as a Detoxification Strategy for Carbon Nanotubes. Carbon N Y 2008,46(3):489–500.

Hamilton RF Jr, Buford M, Xiang C, Wu N, Holian A: NLRP3 inflammasome activation in murine alveolar macrophages and related lung pathology is associated with MWCNT nickel contamination. Inhal Toxicol 2012,24(14):995–1008.

Donaldson K, Murphy FA, Duffin R, Poland CA: Asbestos, carbon nanotubes and the pleural mesothelium: a review of the hypothesis regarding the role of long fibre retention in the parietal pleura, inflammation and mesothelioma. Part Fibre Toxicol 2010, 7: 5.

Nel A, Xia T, Madler L, Li N: Toxic potential of materials at the nanolevel. Science 2006,311(5761):622–627.

Dumortier H, Lacotte S, Pastorin G, Marega R, Wu W, Bonifazi D, Briand JP, Prato M, Muller S, Bianco A: Functionalized carbon nanotubes are non-cytotoxic and preserve the functionality of primary immune cells. Nano Lett 2006,6(7):1522–1528.

Bussy C, Pinault M, Cambedouzou J, Landry MJ, Jegou P, Mayne-L’hermite M, Launois P, Boczkowski J, Lanone S: Critical role of surface chemical modifications induced by length shortening on multi-walled carbon nanotubes-induced toxicity. Part Fibre Toxicol 2012, 9: 46.

Jain S, Thakare VS, Das M, Godugu C, Jain AK, Mathur R, Chuttani K, Mishra AK: Toxicity of Multiwalled Carbon Nanotubes with End Defects Critically Depends on Their Functionalization Density. Chem Res Toxicol 2011,24(11):2028–2039.

Wang Y, Iqbal Z, Mitra S: Rapidly functionalized, water-dispersed carbon nanotubes at high concentration. J Am Chem Soc 2006,128(1):95–99.

Banerjee S, Kahn MGC, Wong SS: Rational chemical strategies for carbon nanotube functionalization. Chem-Eur J 2003,9(9):1899–1908.

Shao L, Tobias G, Salzmann CG, Ballesteros B, Hong SY, Crossley A, Davis BG, Green ML: Removal of amorphous carbon for the efficient sidewall functionalisation of single-walled carbon nanotubes. Chem Commun (Camb) 2007, (47):5090–5092.

Porter DW, Hubbs A, Chen BT, McKinney W, Mercer R, Wolfarth M, Battelli L, Wu N, Sriram K, Leonard S, et al.: Acute pulmonary dose-responses to inhaled multi-walled carbon nanotubes. Nanotoxicology 2012, 7: 1170–1194.

Porter DW, Hubbs AF, Mercer RR, Wu N, Wolfarth MG, Sriram K, Leonard S, Battelli L, Schwegler-Berry D, Friend S, et al.: Mouse pulmonary dose- and time course-responses induced by exposure to multi-walled carbon nanotubes. Toxicology 2010,269(2–3):136–147.

Mercer RR, Hubbs AF, Scabilloni JF, Wang L, Battelli LA, Schwegler-Berry D, Castranova V, Porter DW: Distribution and persistence of pleural penetrations by multi-walled carbon nanotubes. Part Fibre Toxicol 2010,7(1):28.

Tschopp J, Schroder K: NLRP3 inflammasome activation: The convergence of multiple signalling pathways on ROS production? Nat Rev Immunol 2010,10(3):210–215.

Martinon F, Mayor A, Tschopp J: The inflammasomes: guardians of the body. Annu Rev Immunol 2009, 27: 229–265.

Arend WP, Palmer G, Gabay C: IL-1, IL-18, and IL-33 families of cytokines. Immunol Rev 2008, 223: 20–38.

Hamilton RF, Wu N, Porter D, Buford M, Wolfarth M, Holian A: Particle length-dependent titanium dioxide nanomaterials toxicity and bioactivity. Part Fibre Toxicol 2009, 6: 35.

Yazdi AS, Guarda G, Riteau N, Drexler SK, Tardivel A, Couillin I, Tschopp J: Nanoparticles activate the NLR pyrin domain containing 3 (Nlrp3) inflammasome and cause pulmonary inflammation through release of IL-1alpha and IL-1beta. Proc Natl Acad Sci USA 2010,107(45):19449–19454.

Hamilton RF, Xiang C, Li M, Ka I, Yang F, Ma D, Porter D, Wu N, Holian A: Purification and sidewall functionalization of multi-walled carbon nanotubes and resulting bioactivity in two macrophage models. Inhal Toxicol 2013,25(4):199–210.

Dolinay T, Kim YS, Howrylak J, Hunninghake GM, An CH, Fredenburgh L, Massaro AF, Rogers A, Gazourian L, Nakahira K, et al.: Inflammasome-regulated cytokines are critical mediators of acute lung injury. Am J Respir Crit Care Med 2012,185(11):1225–1234.

Johnston HJ, Hutchison GR, Christensen FM, Peters S, Hankin S, Aschberger K, Stone V: A critical review of the biological mechanisms underlying the in vivo and in vitro toxicity of carbon nanotubes: the contribution of physico-chemical characteristics. Nanotoxicology 2010,4(2):207–246.

Sager T, Wolfarth M, Andrew M, Hubbs A, Porter DW, Wu N, Yang F, Hamilton RF Jr, Holian A: Analysis of the effect of multi-walled carbon nanotube surface modification on bioactivity and inflammasome activation. Nanotoxicology 2013. doi: 10.3109/17435390.2013.779757

Bonner JC, Silva RM, Taylor AJ, Brown JM, Hilderbrand SC, Castranova V, Porter D, Elder A, Oberdorster G, Harkema JR, et al.: Interlaboratory evaluation of rodent pulmonary responses to engineered nanomaterials: the NIEHS Nano GO Consortium. Environ Health Perspect 2013,121(6):676–682.

Xia T, Hamilton RF, Bonner JC, Crandall ED, Elder A, Fazlollahi F, Girtsman TA, Kim K, Mitra S, Ntim SA, et al.: Interlaboratory evaluation of in vitro cytotoxicity and inflammatory responses to engineered nanomaterials: the NIEHS Nano GO Consortium. Environ Health Perspect 2013,121(6):683–690.

Stern ST, Adiseshaiah PP, Crist RM: Autophagy and lysosomal dysfunction as emerging mechanisms of nanomaterial toxicity. Part Fibre Toxicol 2012, 9: 20.

Franchi L, Eigenbrod T, Munoz-Planillo R, Nunez G: The inflammasome: a caspase-1-activation platform that regulates immune responses and disease pathogenesis. Nat Immunol 2009,10(3):241–247.

Girtsman TA, Beamer CA, Wu N, Buford M, Holian A: IL-1R signalling is critical for regulation of multi-walled carbon nanotubes-induced acute lung inflammation in C57Bl/6 mice. Nanotoxicology 2012. doi: 10.3109/17435390.2012.744110

Dostert C, Petrilli V, Van Bruggen R, Steele C, Mossman BT, Tschopp J: Innate immune activation through Nalp3 inflammasome sensing of asbestos and silica. Science 2008,320(5876):674–677.

Fenoglio I, Tomatis M, Lison D, Muller J, Fonseca A, Nagy JB, Fubini B: Reactivity of carbon nanotubes: free radical generation or scavenging activity? Free Radic Biol Med 2006,40(7):1227–1233.

Nagai H, Okazaki Y, Chew SH, Misawa N, Yamashita Y, Akatsuka S, Ishihara T, Yamashita K, Yoshikawa Y, Yasui H, et al.: Diameter and rigidity of multiwalled carbon nanotubes are critical factors in mesothelial injury and carcinogenesis. P Natl Acad Sci USA 2011,108(49):E1330-E1338.

Chen YH, Iqbal Z, Mitra S: Microwave-induced controlled purification of single-walled carbon nanotubes without sidewall functionalization. Adv Funct Mater 2007,17(18):3946–3951.

Chen YH, Mitra S: Fast Microwave-Assisted Purification, Functionalization and Dispersion of Multi-Walled Carbon Nanotubes. J Nanosci Nanotechno 2008,8(11):5770–5775.

Ntim SA, Sae-Khow O, Witzmann FA, Mitra S: Effects of polymer wrapping and covalent functionalization on the stability of MWCNT in aqueous dispersions. J Colloid Interf Sci 2011,355(2):383–388.

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