臺灣博碩士論文加值系統

儘管在奈米生物炭的各種應用方面有著令人興奮的潛力和日益增長的興趣，但對其潛在的毒性影響仍存在擔憂。在這裡，我們使用正常的人體外模型來研究上皮接觸納米煙草桔梗熱解生物炭（nTSBs）和肺毒性之間可能存在的毒性潛力。在我們的研究中，在500 oC產生的nTSB被用來分析nTSB對肺的潛在不良影響。結果表明，nTSBs可通過誘導ROS形成而對上皮細胞產生明顯的細胞毒性和遺傳毒性效應。 nTSB暴露也導致激發發炎因子。在暴露於nTSBs損傷的上皮屏障完整性後，跨上皮電阻和緊密連結蛋白的減少。 BEAS-2B細胞中氧化α1-抗胰蛋白酶的增加表明nTSB暴露增加了慢性阻塞性肺病（COPD）的風險。所有這些不良反應都是由高濃度的nTSB引起的。在如此高的濃度下發生的nTSBs的肺部毒性潛力與在更現實的戶外暴露濃度下發生的肺毒性潛力不同。然而，參與製造nTSBs之員工可以長期地暴露於較高濃度的nTSB環境下。因此當其肺部中累積較高nTSB濃度時，即可能增加工作人員罹患肺部疾病之風險。

Despite the exciting potential and growing interest in various applications of nano-biochar, concerns remain regarding their potential toxic effects. Here, we use a normal human in vitro model to investigate the possible toxic potential between epithelial exposure to nano-tobacco stem-pyrolyzed biochars (nTSBs) and pulmonary toxicity. In our study, nTSBs which generate at 500 oC were used to analysis the potential adverse effect of nTSBs on lung. The results showed that nTSBs could cause evident cytotoxic and genotoxic effects towards epithelial cells by inducing ROS formation. nTSBs exposure also led to activation of inflammatory factors. Decreases in transepithelial electrical resistance and zonula occludens proteins after exposure to nTSBs impaired epithelial barrier integrity. Increases in oxidized α1-antitrypsin in BEAS-2B cells suggest that nTSBs exposure increases the risk of chronic obstructive pulmonary disease (COPD). All of these adverse responses were induced by high concentration of nTSBs. The pulmonary toxic potential of nTSBs that occurs at such high concentrations is not the same as that occurring at more realistic outdoor exposure concentrations. However, works involved in the manufacture of nTSBs can be routinely exposed to long-term and high levels of nTSBs. When nTSBs was absorbed into the lung in high enough doses, they may be associated with development of pulmonary disease in workers.

摘要...i
Abstract..ii
誌謝..iii
目錄...iv
表目錄...vi
圖目錄...vii
第一章 緒論...1
1.1 前言...1
1.2 研究動機及目的...1
第二章 文獻回顧...2
2.1 生物炭.... 2
2.2 生物炭應用...2
2.3 生物炭主要成份...2
2.4 生物炭毒性...3
2.5 研究目的...3
第三章 材料與方法...4
3.1 nTSB製備...4
3.2 nTSB之物理化學特性描述...4
3.3 細胞培養...4
3.4 MTT與LDH試驗細胞存活率分析... 5
3.5 以DCFH-DA分析氧化還原狀態...5
3.6 以定量即時反轉錄聚合酶連鎖反應（qRT-RT-PCR）分析氧化還原狀態...5
3.7 以單細胞電泳（彗星）試驗檢驗單股DNA斷裂...6
3.8 以酵素免疫分析法（ELISA）分析DNA損傷、胞內細胞激素、及慢性阻塞性肺炎之預測性生物標記（biomarker）...6
3.9 以免疫轉漬法（immunoblot）分析蛋白質表現...6
3.10 跨表皮細胞電阻（Transepithelial Electrical Resistance，TEER）分析...7
3.11 統計分析...7
第四章 結果與討論...8
4.1 nTSB的物理化學性質...8
4.2 nTSB對BEAS-2B細胞株的細胞毒性效果...10
4.3 nTSB造成的氧化反應...11
4.4 nTSB於BEAS-2B細胞中引發的發炎反應...13
4.5 nTSB處理後造成BEAS-2B細胞的肺部功能喪失...14
第五章 結論...16
參考文獻...17
Extended Abstract...22

1.Zi, W., et al., Optimization of waste tobacco stem expansion by microwave radiation for biomass material using response surface methodology. Journal of the Taiwan Institute of Chemical Engineers, 2013. 44: p. 678-685.
2.Hu, W., et al., Personal and indoor PM2.5 exposure from burning solid fuels in vented and unvented stoves in a rural region of China with a high incidence of lung cancer. Environ Sci Technol, 2014. 48(15): p. 8456-64.
3.Mumba, P.P. and R. Phiri, Environmental impact assessment of tobacco waste disposal. International Journal of Environmental Research, 2008. 2(3): p. 225-230.
4.Cheng, M.T., et al., Particulate matter characteristics during agricultural waste burning in Taichung City, Taiwan. Journal of Hazardous Materials, 2009. 165(1-3): p. 187-192.
5.Lin, Y.H., W. Yan, and K.C. Sheng, Effect of pyrolysis conditions on the characteristics of biochar produced from a tobacco stem. Waste Management & Research, 2016. 34(8): p. 793-801.
6.Liu, Z., P. McNamara, and D. Zitomer, Autocatalytic Pyrolysis of Wastewater Biosolids for Product Upgrading. Environ Sci Technol, 2017. 51(17): p. 9808-9816.
7.Chen, H.P., et al., Physicochemical properties and hygroscopicity of tobacco stem biochar pyrolyzed at different temperatures. Journal of Renewable and Sustainable Energy, 2016. 8(1).
8.Yi, H.H., et al., Preparation of Activated Carbons from Tobacco Stems by Potassium Hydroxide Activation and Phosphine Adsorption. Separation Science and Technology, 2013. 48(5): p. 813-819.
9.Jindo, K., et al., Physical and chemical characterization of biochars derived from different agricultural residues. Biogeosciences, 2014. 11(23): p. 6613-6621.
10.Spokas, K.A., et al., Qualitative analysis of volatile organic compounds on biochar. Chemosphere, 2011. 85(5): p. 869-882.
11.Kambo, H.S. and A. Dutta, A comparative review of biochar and hydrochar in terms of production, physico-chemical properties and applications. Renewable & Sustainable Energy Reviews, 2015. 45: p. 359-378.
12.Xiong, X.N., et al., A review of biochar-based catalysts for chemical synthesis, biofuel production, and pollution control. Bioresource Technology, 2017. 246: p. 254-270.
13.Agegnehu, G., A.K. Srivastava, and M.I. Bird, The role of biochar and biochar-compost in improving soil quality and crop performance: A review. Applied Soil Ecology, 2017. 119: p. 156-170.
14.Huggins, T.M., et al., Graphitic biochar as a cathode electrocatalyst support for microbial fuel cells. Bioresource Technology, 2015. 195: p. 147-153.
15.Wu, X., et al., Long-Life Lithium-Sulfur Battery Derived from Nori-Based Nitrogen and Oxygen Dual-Doped 3D Hierarchical Biochar. Acs Applied Materials & Interfaces, 2017. 9(22): p. 18889-18896.
16.Naghdi, M., et al., A green method for production of nanobiochar by ball milling-optimization and characterization. Journal of Cleaner Production, 2017. 164: p. 1394-1405.
17.Oleszczuk, P., et al., Characterization of nanoparticles of biochars from different biomass. Journal of Analytical and Applied Pyrolysis, 2016. 121: p. 165-172.
18.Joseph, S., et al., Shifting paradigms: development of high-efficiency biochar fertilizers based on nano-structures and soluble components. Carbon Management, 2013. 4(3): p. 323-343.
19.Genovese, M., et al., High capacitive performance of exfoliated biochar nanosheets from biomass waste corn cob. Journal of Materials Chemistry A, 2015. 3(6): p. 2903-2913.
20.Zhou, L., et al., Adsorption Properties of Nano-MnO2-Biochar Composites for Copper in Aqueous Solution. Molecules, 2017. 22(1).
21.Tan, X.F., et al., Biochar-based nano-composites for the decontamination of wastewater: A review. Bioresource Technology, 2016. 212: p. 318-333.
22.Naghdi, M., et al., Pine-wood derived nanobiochar for removal of carbamazepine from aqueous media: Adsorption behavior and influential parameters. Arabian Journal of Chemistry, 2017.
23.He, S.Y., et al., Bioremediation of Wastewater by Iron Oxide-Biochar Nanocomposites Loaded with Photosynthetic Bacteria. Frontiers in Microbiology, 2017. 8.
24.Servin, A.D. and J.C. White, Nanotechnology in agriculture: Next steps for understanding engineered nanoparticle exposure and risk. NanoImpact, 2016. 1: p. 9-12.
25.Freddo, A., C. Cai, and B.J. Reid, Environmental contextualisation of potential toxic elements and polycyclic aromatic hydrocarbons in biochar. Environ Pollut, 2012. 171: p. 18-24.
26.Oleszczuk, P. and M. Koltowski, Changes of total and freely dissolved polycyclic aromatic hydrocarbons and toxicity of biochars treated with various aging processes. Environmental Pollution, 2018. 237: p. 65-73.
27.von Gunten, K., et al., Modified sequential extraction for biochar and petroleum coke: Metal release potential and its environmental implications. Bioresource Technology, 2017. 236: p. 106-110.
28.Koppolu, L., F.A. Agblevor, and L.D. Clements, Pyrolysis as a technique for separating heavy metals from hyperaccumulators. Part II: Lab-scale pyrolysis of synthetic hyperaccumulator biomass. Biomass & Bioenergy, 2003. 25(6): p. 651-663.
29.Oleszczuk, P., I. Jo´sko, and M. Ku´smierz, Biochar properties regarding to contaminants content and ecotoxicological assessment. Journal of Hazardous Materials, 2013. 260: p. 375-382.
30.Lin, C.H., et al., Disparity in the induction of glutathione depletion, ROS formation, poly(ADP-ribose) polymerase-1 activation, and apoptosis by quinonoid derivatives of naphthalene in human cultured cells. Chemico-Biological Interactions, 2007. 165(3): p. 200-210.
31.Lin, P.H., et al., Effects of naphthalene quinonoids on the induction of oxidative DNA damage and cytotoxicity in calf thymus DNA and in human cultured cells. Chemical Research in Toxicology, 2005. 18(8): p. 1262-1270.
32.Keiluweit, M., et al., Solvent-Extractable Polycyclic Aromatic Hydrocarbons in Biochar: Influence of Pyrolysis Temperature and Feedstock. Environmental Science & Technology, 2012. 46(17): p. 9333-9341.
33.Busch, D., et al., Genotoxic and phytotoxic risk assessment of fresh and treated hydrochar from hydrothermal carbonization compared to biochar from pyrolysis. Ecotoxicol Environ Saf, 2013. 97: p. 59-66.
34.Devi, P. and A.K. Saroha, Risk analysis of pyrolyzed biochar made from paper mill effluent treatment plant sludge for bioavailability and eco-toxicity of heavy metals. Bioresource Technology, 2014. 162: p. 308-315.
35.Soudek, P., et al., Characteristics of different types of biochar and effects on the toxicity of heavy metals to germinating sorghum seeds. Journal of Geochemical Exploration, 2017. 182: p. 157-165.
36.Lyu, H.H., et al., Effect of pyrolysis temperature on potential toxicity of biochar if applied to the environment. Environmental Pollution, 2016. 218: p. 1-7.
37.Rizwan, M., et al., Mechanisms of biochar-mediated alleviation of toxicity of trace elements in plants: a critical review. Environmental Science and Pollution Research, 2016. 23(3): p. 2230-2248.
38.John, A.C., et al., Emissions and Possible Environmental Implication of Engineered Nanomaterials (ENMs) in the Atmosphere. Atmosphere, 2017. 8(5).
39.Shaaban, A., et al., Characterization of biochar derived from rubber wood sawdust through slow pyrolysis on surface porosities and functional groups. Procedia Engineering, 2013. 68: p. 365-371.
40.Seaton, A., et al., Nanoparticles, human health hazard and regulation. Journal of the Royal Society Interface, 2010. 7: p. S119-S129.
41.Sun, Z.L., et al., Nano zerovalent iron particles induce pulmonary and cardiovascular toxicity in an in vitro human co-culture model. Nanotoxicology, 2016. 10(7): p. 881-890.
42.Yan, J., et al., Consecutive evaluation of graphene oxide and reduced graphene oxide nanoplatelets immunotoxicity on monocytes. Colloids Surf B Biointerfaces, 2017. 153: p. 300-309.
43.Otero-Gonzalez, L., et al., Application and Validation of an Impedance-Based Real Time Cell Analyzer to Measure the Toxicity of Nanoparticles Impacting Human Bronchial Epithelial Cells. Environmental Science & Technology, 2012. 46(18): p. 10271-10278.
44.Yan, J.Y., et al., Industrial PM2.5 cause pulmonary adverse effect through RhoA/ROCK pathway. Science of the Total Environment, 2017. 599: p. 1658-1666.
45.Yang, L.Y., et al., Polycyclic aromatic hydrocarbons are associated with increased risk of chronic obstructive pulmonary disease during haze events in China. Science of the Total Environment, 2017. 574: p. 1649-1658.
46.Ossola, J.O. and M.L. Tomaro, Heme oxygenase induction by UVA radiation. A response to oxidative stress in rat liver. Int J Biochem Cell Biol, 1998. 30(2): p. 285-92.
47.Birben, E., et al., Oxidative stress and antioxidant defense. World Allergy Organ J, 2012. 5(1): p. 9-19.
48.Lin, C.H., et al., Cd/Se/Te-based quantum dot 705 modulated redox homeostasis with hepatotoxicity in mice. Nanotoxicology, 2011. 5(4): p. 650-663.
49.Griffith, O.W., Biologic and pharmacologic regulation of mammalian glutathione synthesis. Free Radic Biol Med, 1999. 27(9-10): p. 922-35.
50.Danielsen, P.H., et al., DNA damage in rats after a single oral exposure to diesel exhaust particles. Mutat Res, 2008. 637(1-2): p. 49-55.
51.Caramori, G., et al., Mechanisms involved in lung cancer development in COPD. International Journal of Biochemistry & Cell Biology, 2011. 43(7): p. 1030-1044.
52.You, K., et al., Hyperoxia disrupts pulmonary epithelial barrier in newborn rats via the deterioration of occludin and ZO-1. Respir Res, 2012. 13: p. 36.
53.Zhang, Y.L., et al., Effects of chronic ethanol ingestion on tight junction proteins and barrier function of alveolar epithelium in the rat. Shock, 2007. 28(2): p. 245-52.
54.Ferrara, A., Chronic obstructive pulmonary disease. Radiol Technol, 2011. 82(3): p. 245-63.
55.Alam, S., et al., Oxidation of Z alpha1-antitrypsin by cigarette smoke induces polymerization: a novel mechanism of early-onset emphysema. Am J Respir Cell Mol Biol, 2011. 45(2): p. 261-9.
56.Li, Z., et al., Oxidized {alpha}1-antitrypsin stimulates the release of monocyte chemotactic protein-1 from lung epithelial cells: potential role in emphysema. Am J Physiol Lung Cell Mol Physiol, 2009. 297(2): p. L388-400.