Fluorescent Nanodiamonds for Biomedical Applications: Cytotoxicity, In vitro and In vivo studies

簡易檢索 / 詳目顯示

回結果列表

研究生：	蜜瑪拉 VAIJAYANTHIMALA.V
論文名稱：	Fluorescent Nanodiamonds for Biomedical Applications: Cytotoxicity, In vitro and In vivo studies 螢光奈米鑽石於生醫上之應用: 毒性、活體外與活體內之研究
指導教授：	張煥正 Chang, Huan-Cheng 倪其焜 Ni, Chi-Kung
口試委員:	張煥正 Chang, Huan-Cheng 倪其焜 Ni, Chi-Kung 韓肇中 Han, Chau-Chung 李仲良 Li, Chung-Leung 黃國柱 Hwang, Kuo-Chu
學位類別：	博士 Doctor
系所名稱：	理學院 - 化學系 Department of Chemistry
論文出版年：	2011
畢業學年度：	100
語文別：	英文
論文頁數：	125
中文關鍵詞：	螢光穩定性、螢光奈米鑽石、細胞倍增時間、活體幹細胞、活體影像
外文關鍵詞：	Fluorescent Nanodiamonds, Flowcytometry, Endocytosis, Exocytosis, Invivo imaging
相關次數：	點閱：4 下載：0
分享至:	分享至facebook 分享至twitter

查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

Fluorescent nanodiamond is a novel carbon based nanoparticle with a variety of important and unique properties such as photostability, biocompatibility, facile surface functionalizability making it as a novel nanoparticle for both in vitro and in vivo applications. Nanodiamond upon high energy ion beam irradiation and subsequent annealing at high temperature, results in the formation of (N-V)0 and (N-V)- defect centers which emit far red fluorescence under visible light excitation. Aforementioned properties make FND as a robust nanoparticle for cell labeling, cell tracking, in vitro and in vivo imaging applications.

In brief, this research begins with the cytotoxicity studies of FNDs in various cell lines. Our studies suggested that FNDs are biocompatible to different cell lines including HeLa, 3T3-L1 and 489-2.1 cells. Further studies showed that the FND cellular uptake is by energy dependent, clathrin mediated endocytosis. Based on the cell doubling time measurements, we found ~10% of FNDs are exocytosed both in short term as well as in long term. In addition to the fluorescence, light scattering also serve as an important parameter that could be utilized during cell sorting. Furthermore, FNDs can serve as an alternative probe to the commonly used dye like CFSE. We have demonstrated the feasibility of using FNDs for in vivo imaging. Injected FNDs accumulate in axillary lymph node and we also developed a new powerful methodology for the quantitation of FNDs in tissues and organs. Finally, we have also demonstrated that FND can be used as a fluorescent marker for in vivo stem cell tracking. In conclusion, FND can serve as an ideal nanoparticle for different biological applications.

螢光奈米鑽石為碳原子組成的新穎材料，許多重要且獨特的性質，如：螢光穩定性、生物相容性、表面修飾多元性等，使其成為生物體外及活體影像應用的重要材料。經過高能離子束靶擊及高溫淬火的奈米鑽石內，產生(N-V) 0、(N-V)- 缺陷中心，以可見光激發此類缺陷中心，可放出近紅外波長的螢光。上述性質使螢光奈米鑽石具有細胞標記、細胞追蹤及生物體外及活體影像應用的潛力。此篇論文首先研究螢光奈米鑽石對不同細胞株的細胞毒性，結果顯示螢光奈米鑽石在3T3-L1, 489-2.1 細胞株內生物相容性好; 進一步研究顯示, 細胞藉由網格蛋白輔助 (clathrin mediated）吞噬螢光奈米鑽石；細胞倍增時間（cell doubling time）實驗中，約百分之十的螢光奈米鑽石在短期及長時間內被細胞排出，除了螢光之外，散射光亦是細胞分選的重要參數，本實驗證明螢光奈米鑽石亦可做細胞分選的探針，與常用的有機染料CFSE具有相似功能；螢光奈米鑽石亦展現活體影像的應用潛力，注射入老鼠的螢光奈米鑽石累積於輔助淋巴結中，並由本實驗發展的方法進行組織及器官內螢光奈米鑽石的定量。最後，螢光奈米鑽石亦被證實可做為活體幹細胞追蹤的螢光標籤。總結，螢光奈米鑽石為生物應用的理想材料。

   Fluorescent Nanodiamonds    1
1 Introduction    1
2 Classification of Diamonds    1
3 Synthesis of HPHT Diamond    2
4 Vacancy related defect centers    4
5 Preparation of fluorescent nanodiamonds    5
6 Optical characterization    6
7 References    10
   Nanodiamonds-Literature review    13
1 Introduction    13
2 Biocompatibility    15
3 Surface modification    17
4 Photoluminescence    21
5 Conclusion    27
6 Future perspective    27
7 References    28
   Cytotoxicity studies of FNDs    35
1 Introduction    36
2 Experimental Methods    37
3 Results and Discussion    41
4 Summary    53
5 References    53
   Cellular uptake mechanism of FNDs    55
1 Introduction    56
2 Experimental methods    57
3 Results and Discussion    59
4 Summary    66
5 References    67
   Exocytosis and in vitro applications of FND    68
1. Introduction    69
2 Experimental Section    71
3 Results and Discussion    76
4 Summary    94
5 References    94
   Fluorescent nanodiamonds in mouse and rat models:Optical imaging and histopathological analysis    97
1 Introduction    98
2 Experimental Section    100
3 Results and discussion    103
4 Summary    111
5 References    111
   Fluorescent Nanodiamonds for in vivo tracking    113
1 Introduction    114
2 Experimental section    115
3 Results and discussion    119
4 Summary    125

                                

1. Freitas Jr RA: Nanomedicine (Volume IIA): Biocompatibility. Landes Bioscience, Georgetown, TX, USA (2003).
2. Burns RC, Davies GJ: Properties of Natural and Synthetic Diamond. Field JE (Ed.). Academic Press, London, UK 395–422 (1992).
3. Moustakas TD: Synthetic Diamond: Emerging CVD Science and Technology. Spear KE, Dismukes JP (Eds). John Wiley, New York, USA 145–192 (1994).
4. Dion I, Baquey C, Monties JR: Diamond – the biomaterial of the 21st century? Int. J. Artif. Organs 16, 623–627 (1993).
5. Dahl, J. E.; Liu, S. G.; Carlson, R. M. K., Isolation and Structure of Higher Diamondoids, Nanometer-Sized Diamond Molecules. Science 2003, 299 (5603), 96-99.
6. Dahl, J. E. P.; Moldowan, J. M.; Peakman, T. M.; Clardy, J. C.; Lobkovsky, E.; Olmstead, M. M.; May, P. W.; Davis, T. J.; Steeds, J. W.; Peters, K. E.; Pepper, A.; Ekuan, A.; Carlson, R. M. K., Isolation and Structural Proof of the Large Diamond Molecule, Cyclohexamantane (C26H30). Angewandte Chemie International Edition 2003, 42 (18), 2040-2044.
7. Krueger, A., The structure and reactivity of nanoscale diamond. Journal of Materials Chemistry 2008, 18 (13), 1485-1492.
8. Yu, S.-J.; Kang, M.-W.; Chang, H.-C.; Chen, K.-M.; Yu, Y.-C., Bright Fluorescent Nanodiamonds: No Photobleaching and Low Cytotoxicity. Journal of the American Chemical Society 2005, 127 (50), 17604-17605.
9. Fu, C.-C.; Lee, H.-Y.; Chen, K.; Lim, T.-S.; Wu, H.-Y.; Lin, P.-K.; Wei, P.-K.; Tsao, P.-H.; Chang, H.-C.; Fann, W., Characterization and application of single fluorescent nanodiamonds as cellular biomarkers. Proceedings of the National Academy of Sciences 2007, 104 (3), 727-732.
10. Chang, Y.-R.; Lee, H.-Y.; Chen, K.; Chang, C.-C.; Tsai, D.-S.; Fu, C.-C.; Lim, T.-S.; Tzeng, Y.-K.; Fang, C.-Y.; Han, C.-C.; Chang, H.-C.; Fann, W., Mass production and dynamic imaging of fluorescent nanodiamonds. Nat Nano 2008, 3 (5), 284-288.
11. Schwertfeger, H.; Fokin, A. A.; Schreiner, P. R., Diamonds are a Chemist's Best Friend: Diamondoid Chemistry Beyond Adamantane. Angewandte Chemie International Edition 2008, 47 (6), 1022-1036.
12. Nebel, C. E.; Shin, D.; Rezek, B.; Tokuda, N.; Uetsuka, H.; Watanabe, H., Diamond and biology. Journal of The Royal Society Interface 2007, 4 (14), 439-461.
13. Holt, K. B., Diamond at the Nanoscale: Applications of Diamond Nanoparticles from Cellular Biomarkers to Quantum Computing. Philosophical Transactions: Mathematical, Physical and Engineering Sciences 2007, 365 (1861), 2845-2861.
14. Krueger, A., New Carbon Materials: Biological Applications of Functionalized Nanodiamond Materials. Chemistry – A European Journal 2008, 14 (5), 1382-1390.
15. Manilenko VV: Ultrananocrystalline Diamond: Synthesis, Properties and Applications. Shenderova OA, Gruen DM (Eds). William Andrew, New York, USA 335–345 (2006).
16. Yang, W.; Auciello, O.; Butler, J. E.; Cai, W.; Carlisle, J. A.; Gerbi, J. E.; Gruen, D. M.; Knickerbocker, T.; Lasseter, T. L.; Russell, J. N.; Smith, L. M.; Hamers, R. J., DNA-modified nanocrystalline diamond thin-films as stable, biologically active substrates. Nat Mater 2003, 2 (1), 63-63.
17. Hartl, A.; Schmich, E.; Garrido, J. A.; Hernando, J.; Catharino, S. C. R.; Walter, S.; Feulner, P.; Kromka, A.; Steinmuller, D.; Stutzmann, M., Protein-modified nanocrystalline diamond thin films for biosensor applications. Nat Mater 2004, 3 (10), 736-742.
18. Osawa, E., Recent progress and perspectives in single-digit nanodiamond. Diamond and Related Materials 2007, 16 (12), 2018-2022.
19. Huang, H.; Pierstorff, E.; Osawa, E.; Ho, D., Active Nanodiamond Hydrogels for Chemotherapeutic Delivery. Nano Letters 2007, 7 (11), 3305-3314.
20. Davies, G.; Hamer, M. F., Optical Studies of the 1.945 eV Vibronic Band in Diamond. Proceedings of the Royal Society of London. Series A, Mathematical and Physical Sciences 1976, 348 (1653), 285-298.
21. Taik Lim, Y.; Kim, S.; Nakayama, A.; Stott, N.; Bawendi, M.; Frangioni, J., Selection of Quantum Dot Wavelengths for Biomedical Assays and Imaging. Molecular Imaging 2003, 2 (1).
22. Brannon-Peppas, L.; Blanchette, J. O., Nanoparticle and targeted systems for cancer therapy. Advanced Drug Delivery Reviews 2004, 56 (11), 1649-1659.
23. Kawasaki, E. S.; Player, A., Nanotechnology, nanomedicine, and the development of new, effective therapies for cancer. Nanomedicine: Nanotechnology, Biology and Medicine 2005, 1 (2), 101-109.
24. Zhang, L.; Gu, F. X.; Chan, J. M.; Wang, A. Z.; Langer, R. S.; Farokhzad, O. C., Nanoparticles in Medicine: Therapeutic Applications and Developments. Clinical Pharmacology and Therapeutics 2007, 83 (5), 761-769.
25. Colvin, V. L., The potential environmental impact of engineered nanomaterials. Nature Biotechnology 2003, 21 (10), 1166-1170.
26. Jia, G.; Wang, H.; Yan, L.; Wang, X.; Pei, R.; Yan, T.; Zhao, Y.; Guo, X., Cytotoxicity of Carbon Nanomaterials: Single-Wall Nanotube, Multi-Wall Nanotube, and Fullerene. Environmental Science & Technology 2005, 39 (5), 1378-1383.
27. Liu, K.-K.; et al., Biocompatible and detectable carboxylated nanodiamond on human cell. Nanotechnology 2007, 18 (32), 325102.
28. Schrand, A. M.; Huang, H.; Carlson, C.; Schlager, J. J.; Ōsawa, E.; Hussain, S. M.; Dai, L., Are Diamond Nanoparticles Cytotoxic? The Journal of Physical Chemistry B 2006, 111 (1), 2-7.
29. Schrand, A. M.; Dai, L.; Schlager, J. J.; Hussain, S. M.; Osawa, E., Differential biocompatibility of carbon nanotubes and nanodiamonds. Diamond and Related Materials 2007, 16 (12), 2118-2123.
30. Ushizawa, K.; Sato, Y.; Mitsumori, T.; Machinami, T.; Ueda, T.; Ando, T., Covalent immobilization of DNA on diamond and its verification by diffuse reflectance infrared spectroscopy. Chemical Physics Letters 2002, 351 (1-2), 105-108.
31. Huang, L. C. L.; Chang, H.-C., Adsorption and Immobilization of Cytochrome c on Nanodiamonds. Langmuir 2004, 20 (14), 5879-5884.
32. Kruger, A.; Liang, Y.; Jarre, G.; Stegk, J., Surface functionalisation of detonation diamond suitable for biological applications. Journal of Materials Chemistry 2006, 16 (24), 2322-2328.
33. Chung, P. H.; Perevedentseva, E.; Tu, J. S.; Chang, C. C.; Cheng, C. L., Spectroscopic study of bio-functionalized nanodiamonds. Diamond and Related Materials 15 (4-8), 622-625.
34. Osswald, S.; Yushin, G.; Mochalin, V.; Kucheyev, S. O.; Gogotsi, Y., Control of sp2/sp3 Carbon Ratio and Surface Chemistry of Nanodiamond Powders by Selective Oxidation in Air. Journal of the American Chemical Society 2006, 128 (35), 11635-11642.
35. Nguyen, T. T.-B.; Chang, H.-C.; Wu, V. W.-K., Adsorption and hydrolytic activity of lysozyme on diamond nanocrystallites. Diamond and Related Materials 2007, 16 (4-7), 872-876.
36. Kong, X. L.; Huang, L. C. L.; Hsu, C. M.; Chen, W. H.; Han, C. C.; Chang, H. C., High-Affinity Capture of Proteins by Diamond Nanoparticles for Mass Spectrometric Analysis. Analytical Chemistry 2004, 77 (1), 259-265.
37. Chen, W.-H.; Lee, S.-C.; Sabu, S.; Fang, H.-C.; Chung, S.-C.; Han, C.-C.; Chang, H.-C., Solid-Phase Extraction and Elution on Diamond (SPEED): A Fast and General Platform for Proteome Analysis with Mass Spectrometry. Analytical Chemistry 2006, 78 (12), 4228-4234.
38. Sabu, S.; Yang, F.-C.; Wang, Y.-S.; Chen, W.-H.; Chou, M.-I.; Chang, H.-C.; Han, C.-C., Peptide analysis: Solid phase extraction-elution on diamond combined with atmospheric pressure matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry. Analytical Biochemistry 2007, 367 (2), 190-200.
39. Tzeng, Y.-K.; Chang, C.-C.; Huang, C.-N.; Wu, C.-C.; Han, C.-C.; Chang, H.-C., Facile MALDI-MS Analysis of Neutral Glycans in NaOH-Doped Matrixes: Microwave-Assisted Deglycosylation and One-Step Purification with Diamond Nanoparticles. Analytical Chemistry 2008, 80 (17), 6809-6814.
40. Kong, X.; Huang, L. C. L.; Liau, S. C. V.; Han, C.-C.; Chang, H.-C., Polylysine-Coated Diamond Nanocrystals for MALDI-TOF Mass Analysis of DNA Oligonucleotides. Analytical Chemistry 2005, 77 (13), 4273-4277.
41. Kossovsky, N.; Gelman, A.; Hnatyszyn, H. J.; Rajguru, S.; Garrell, R. L.; Torbati, S.; Freitas, S. S. F.; Chow, G.-M., Surface-Modified Diamond Nanoparticles as Antigen Delivery Vehicles. Bioconjugate Chemistry 1995, 6 (5), 507-511.
42. Cheng, C. Y.; Perevedentseva, E.; Tu, J. S.; Chung, P. H.; Cheng, C. L.; Liu, K. K.; Chao, J. I.; Chen, P. H.; Chang, C. C., Direct and in vitro observation of growth hormone receptor molecules in A549 human lung epithelial cells by nanodiamond labeling. Appl. Phys. Lett. 2007, 90 (16).
43. Vial, S.; Mansuy, C.; Sagan, S.; Irinopoulou, T.; Burlina, F.; Boudou, J.-P.; Chassaing, G.; Lavielle, S., Peptide-Grafted Nanodiamonds: Preparation, Cytotoxicity and Uptake in Cells. ChemBioChem 2008, 9 (13), 2113-2119.
44. Krueger, A.; Stegk, J.; Liang, Y.; Lu, L.; Jarre, G., Biotinylated Nanodiamond: Simple and Efficient Functionalization of Detonation Diamond. Langmuir 2008, 24 (8), 4200-4204.
45. Yeap, W. S.; Tan, Y. Y.; Loh, K. P., Using Detonation Nanodiamond for the Specific Capture of Glycoproteins. Analytical Chemistry 2008, 80 (12), 4659-4665.
46. Liu, K.-K.; Mei-Fang, C.; Chen, P.-Y.; Lee, T. J. F.; Cheng, C.-L.; Chang, C.-C.; Ho, Y.-P.; Chao, J.-I., Alpha-bungarotoxin binding to target cell in a developing visual system by carboxylated nanodiamond. Nanotechnology 2008, 19 (20).
47. Taton, T. A., Nanostructures as tailored biological probes. Trends in Biotechnology 2002, 20 (7), 277-279.
48. Giepmans, B. N. G.; Adams, S. R.; Ellisman, M. H.; Tsien, R. Y., The Fluorescent Toolbox for Assessing Protein Location and Function. Science 2006, 312 (5771), 217-224.
49. Zhang, J.; Campbell, R. E.; Ting, A. Y.; Tsien, R. Y., Creating new fluorescent probes for cell biology. Nat Rev Mol Cell Biol 2002, 3 (12), 906-918.
50. Michalet, X.; Pinaud, F. F.; Bentolila, L. A.; Tsay, J. M.; Doose, S.; Li, J. J.; Sundaresan, G.; Wu, A. M.; Gambhir, S. S.; Weiss, S., Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics. Science 2005, 307 (5709), 538-544.
51. Derfus, A. M.; Chan, W. C. W.; Bhatia, S. N., Probing the Cytotoxicity of Semiconductor Quantum Dots. Nano Letters 2003, 4 (1), 11-18.
52. Cho, S. J.; Maysinger, D.; Jain, M.; Röder, B.; Hackbarth, S.; Winnik, F. M., Long-Term Exposure to CdTe Quantum Dots Causes Functional Impairments in Live Cells. Langmuir 2007, 23 (4), 1974-1980.
53. Billinton, N.; Knight, A. W., Seeing the Wood through the Trees: A Review of Techniques for Distinguishing Green Fluorescent Protein from Endogenous Autofluorescence. Analytical Biochemistry 2001, 291 (2), 175-197.
54. Mansfield, J. R.; Gossage, K. W.; Hoyt, C. C.; Levenson, R. M., Autofluorescence removal, multiplexing, and automated analysis methods for in-vivo fluorescence imaging. Journal of Biomedical Optics 2005, 10 (4).
55. Treussart, F.; Jacques, V.; Wu, E.; Gacoin, T.; Grangier, P.; Roch, J. F., Photoluminescence of single colour defects in 50 nm diamond nanocrystals. Physica B: Condensed Matter 2006, 376-377, 926-929.
56. Neugart, F.; Zappe, A.; Jelezko, F.; Tietz, C.; Boudou, J. P.; Krueger, A.; Wrachtrup, J., Dynamics of Diamond Nanoparticles in Solution and Cells. Nano Letters 2007, 7 (12), 3588-3591.
57. Wee, T.-L.; Mau, Y.-W.; Fang, C.-Y.; Hsu, H.-L.; Han, C.-C.; Chang, H.-C., Preparation and characterization of green fluorescent nanodiamonds for biological applications. Diamond and Related Materials 2009, 18 (2-3), 567-573.
58. Davies, G.; Lawson, S. C.; Collins, A. T.; Mainwood, A.; Sharp, S. J., Vacancy-related centers in diamond. Physical Review B 1992, 46 (20), 13157.
59. Smith, B. R.; Niebert, M.; Plakhotnik, T.; Zvyagin, A. V., Transfection and imaging of diamond nanocrystals as scattering optical labels. Journal of Luminescence 2007, 127 (1), 260-263.
60. Chao, J.-I.; Perevedentseva, E.; Chung, P.-H.; Liu, K.-K.; Cheng, C.-Y.; Chang, C.-C.; Cheng, C.-L., Nanometer-Sized Diamond Particle as a Probe for Biolabeling. Biophysical journal 2007, 93 (6), 2199-2208.
61. Cheng, C. L.; Perevedentseva, E.; Cheng, C. Y.; Chung, P. H.; Tu, J. S.; Hsieh, Y. H., The interaction of the protein lysozyme with bacteria E-coli observed using nanodiamond labelling. Nanotechnology 2007, 18 (31).
62. Sanhai, W. R.; Sakamoto, J. H.; Canady, R.; Ferrari, M., Seven challenges for nanomedicine. Nat Nano 2008, 3 (5), 242-244.
63. Marcon, L.; Riquet, F.; Vicogne, D.; Szunerits, S.; Bodart, J.-F.; Boukherroub, R., Cellular and in vivo toxicity of functionalized nanodiamond in Xenopus embryos. Journal of Materials Chemistry 2010, 20 (37), 8064-8069.
64. Xing, Y.; Xiong, W.; Zhu, L.; Osawa, E.; Hussin, S.; Dai, L., DNA Damage in Embryonic Stem Cells Caused by Nanodiamonds. ACS Nano 2011, 5 (3), 2376-2384.
65. Yuan, Y.; Chen, Y.; Liu, J.-H.; Wang, H.; Liu, Y., Biodistribution and fate of nanodiamonds in vivo. Diamond and Related Materials 2009, 18 (1), 95-100.
66. Mohan, N.; Chen, C.-S.; Hsieh, H.-H.; Wu, Y.-C.; Chang, H.-C., In Vivo Imaging and Toxicity Assessments of Fluorescent Nanodiamonds in Caenorhabditis elegans. Nano Letters 2010, 10 (9), 3692-3699.
67. Bakowicz, K.; Mitura, S., Biocompatibility of NCD. Journal of Wide Bandgap Materials 2002, 9 (4), 261-272.
68. Puzyr, A. P.; Baron, A. V.; Purtov, K. V.; Bortnikov, E. V.; Skobelev, N. N.; Mogilnaya, O. A.; Bondar, V. S., Nanodiamonds with novel properties: A biological study. Diamond and Related Materials 2007, 16 (12), 2124-2128.
69. Wang, H. F.; Yuan, Y.; Wang, X.; Jia, G.; Liu, J. H.; Wang, T. C.; Gu, Y. Q.; Yang, S. T.; Zhen, S.; Liu, Y. F., Pulmonary toxicity and translocation of nanodiamonds in mice. Diamond and Related Materials 2010, 19 (4), 291-299.

全文公開日期本全文未授權公開 (校內網路)
全文公開日期本全文未授權公開 (校外網路)

簡易檢索 / 詳目顯示

相關論文