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DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳嘉文(Chia-Wen (Kevin) | |
dc.contributor.author | Ya-Heui Yang | en |
dc.contributor.author | 楊雅惠 | zh_TW |
dc.date.accessioned | 2021-05-20T20:59:50Z | - |
dc.date.available | 2016-07-27 | |
dc.date.available | 2021-05-20T20:59:50Z | - |
dc.date.copyright | 2011-07-27 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-07-22 | |
dc.identifier.citation | 1. Zhang, C., et al., Self-activated luminescent and mesoporous strontium hydroxyapatite nanorods for drug delivery. Biomaterials, 2010. 31(12): p. 3374-3383.
2. Slowing, I.I., et al., Mesoporous silica nanoparticles as controlled release drug delivery and gene transfection carriers. Advanced Drug Delivery Reviews, 2008. 60(11): p. 1278-1288. 3. Kester, M., et al., Calcium Phosphate Nanocomposite Particles for In Vitro Imaging and Encapsulated Chemotherapeutic Drug Delivery to Cancer Cells. Nano Letters, 2008. 8(12): p. 4116-4121. 4. Morgan, T.T., et al., Encapsulation of Organic Molecules in Calcium Phosphate Nanocomposite Particles for Intracellular Imaging and Drug Delivery. Nano Letters, 2008. 8(12): p. 4108-4115. 5. Yang, J., et al., Hollow Silica Nanocontainers as Drug Delivery Vehicles. Langmuir, 2008. 24(7): p. 3417-3421. 6. Son, S.J., X. Bai, and S.B. Lee, Inorganic hollow nanoparticles and nanotubes in nanomedicine: Part 1. Drug/gene delivery applications. Drug Discovery Today, 2007. 12(15-16): p. 650-656. 7. Sanvicens, N. and M.P. Marco, Multifunctional nanoparticles - properties and prospects for their use in human medicine. Trends in Biotechnology, 2008. 26(8): p. 425-433. 8. Riehemann, K., et al., Nanomedicine—Challenge and Perspectives. Angewandte Chemie International Edition, 2009. 48(5): p. 872-897. 9. Doshi, N. and S. Mitragotri, Designer Biomaterials for Nanomedicine. Advanced Functional Materials, 2009. 19(24): p. 3843-3854. 10. Son, S.J., X. Bai, and S.B. Lee, Inorganic hollow nanoparticles and nanotubes in nanomedicine: Part 2: Imaging, diagnostic, and therapeutic applications. Drug Discovery Today, 2007. 12(15-16): p. 657-663. 11. Ow, H., et al., Bright and Stable Core−Shell Fluorescent Silica Nanoparticles. Nano Letters, 2004. 5(1): p. 113-117. 12. Huo, Q., et al., A New Class of Silica Cross-Linked Micellar Core−Shell Nanoparticles. Journal of the American Chemical Society, 2006. 128(19): p. 6447-6453. 13. Panyam, J. and V. Labhasetwar, Biodegradable nanoparticles for drug and gene delivery to cells and tissue. Advanced Drug Delivery Reviews, 2003. 55(3): p. 329-347. 14. Brigger, I., C. Dubernet, and P. Couvreur, Nanoparticles in cancer therapy and diagnosis. Advanced Drug Delivery Reviews, 2002. 54(5): p. 631-651. 15. Singh, R. and J.W. Lillard Jr, Nanoparticle-based targeted drug delivery. Experimental and Molecular Pathology, 2009. 86(3): p. 215-223. 16. Zhang, L., et al., Nanoparticles in Medicine: Therapeutic Applications and Developments. Clin Pharmacol Ther, 2007. 83(5): p. 761-769. 17. Torchilin, V.P., Multifunctional nanocarriers. Advanced Drug Delivery Reviews, 2006. 58(14): p. 1532-1555. 18. Letchford, K. and H. Burt, A review of the formation and classification of amphiphilic block copolymer nanoparticulate structures: micelles, nanospheres, nanocapsules and polymersomes. European Journal of Pharmaceutics and Biopharmaceutics, 2007. 65(3): p. 259-269. 19. Xu, Z.P., et al., Inorganic nanoparticles as carriers for efficient cellular delivery. Chemical Engineering Science, 2006. 61(3): p. 1027-1040. 20. Fendler, J.H. and A. Romero, Liposomes as drug carriers. Life Sciences, 1977. 20(7): p. 1109-1120. 21. Jones, M.-C. and J.-C. Leroux, Polymeric micelles - a new generation of colloidal drug carriers. European Journal of Pharmaceutics and Biopharmaceutics, 1999. 48(2): p. 101-111. 22. Uskoković, V. and D.P. Uskoković, Nanosized hydroxyapatite and other calcium phosphates: Chemistry of formation and application as drug and gene delivery agents. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2011. 96B(1): p. 152-191. 23. de Leeuw, N.H., Computer simulations of structures and properties of the biomaterial hydroxyapatite. Journal of Materials Chemistry, 2010. 20(26): p. 5376-5389. 24. Elliott, J.C., P.E. Mackie, and R.A. Young, Monoclinic Hydroxyapatite. Science, 1973. 180(4090): p. 1055-1057. 25. Espanol, M., et al., Investigation of the hydroxyapatite obtained as hydrolysis product of [small alpha]-tricalcium phosphate by transmission electron microscopy. CrystEngComm, 2010. 12(10): p. 3318-3326. 26. LeGeros, R.Z., Calcium Phosphate-Based Osteoinductive Materials. Chemical Reviews, 2008. 108(11): p. 4742-4753. 27. Ye, F., et al., Polymeric micelle-templated synthesis of hydroxyapatite hollow nanoparticles for a drug delivery system. Acta Biomaterialia, 2010. 6(6): p. 2212-2218. 28. He, W., et al., A Size-controlled Synthesis of Hollow Apatite Nanospheres at Water–Oil Interfaces. Chemistry Letters, 2010. 39(7): p. 674-675. 29. Itatani, K., et al., Preparation of submicrometer-sized porous spherical hydroxyapatite agglomerates by ultrasonic spray pyrolysis technique. Journal of the Ceramic Society of Japan, 2010. 118(1378): p. 462-466. 30. Ma, M.-Y., et al., Nanostructured porous hollow ellipsoidal capsules of hydroxyapatite and calcium silicate: preparation and application in drug delivery. Journal of Materials Chemistry, 2008. 18(23): p. 2722-2727. 31. Wang, Y., et al., Fast precipitation of uniform CaCO3 nanospheres and their transformation to hollow hydroxyapatite nanospheres. Journal of Colloid and Interface Science, 2010. 352(2): p. 393-400. 32. Espinosa, E., et al., Classification of anticancer drugs--a new system based on therapeutic targets. Cancer Treatment Reviews, 2003. 29(6): p. 515-523. 33. Wu, X.-Z., A new classification system of anticancer drugs - Based on cell biological mechanisms. Medical Hypotheses, 2006. 66(5): p. 883-887. 34. Gewirtz, D.A., A critical evaluation of the mechanisms of action proposed for the antitumor effects of the anthracycline antibiotics adriamycin and daunorubicin. Biochemical Pharmacology, 1999. 57(7): p. 727-741. 35. Keizer, H.G., et al., Doxorubicin (adriamycin): A critical review of free radical-dependent mechanisms of cytotoxicity. Pharmacology & Therapeutics, 1990. 47(2): p. 219-231. 36. Binaschi, M., et al., In Vivo Site Specificity and Human Isoenzyme Selectivity of Two Topoisomerase II-poisoning Anthracyclines. Cancer Research, 2000. 60(14): p. 3770-3776. 37. Froelich-Ammon, S.J. and N. Osheroff, Topoisomerase Poisons: Harnessing the Dark Side of Enzyme Mechanism. Journal of Biological Chemistry, 1995. 270(37): p. 21429-21432. 38. Binaschi, M., et al., Relationship between Lethal Effects and Topoisomerase II-Mediated Double-Stranded DNA Breaks Produced by Anthracyclines with Different Sequence Specificity. Molecular Pharmacology, 1997. 51(6): p. 1053-1059. 39. Capranico, G., et al., Markedly Reduced Levels of Anthracycline-induced DNA Strand Breaks in Resistant P388 Leukemia Cells and Isolated Nuclei. Cancer Research, 1987. 47(14): p. 3752-3756. 40. Zunino, F., G. Pratesi, and P. Perego, Role of the sugar moiety in the pharmacological activity of anthracyclines: development of a novel series of disaccharide analogs. Biochemical Pharmacology, 2001. 61(8): p. 933-938. 41. Capranico, G., et al., Role of DNA Breakage in Cytotoxicity of Doxorubicin, 9-Deoxydoxorubicin, and 4-Demethyl-6-deoxydoxorubicin in Murine Leukemia P388 Cells. Cancer Research, 1989. 49(8): p. 2022-2027. 42. Capranico, G., E. Butelli, and F. Zunino, Change of the Sequence Specificity of Daunorubicin-stimulated Topoisomerase II DNA Cleavage by Epimerization of the Amino Group of the Sugar Moiety. Cancer Research, 1995. 55(2): p. 312-317. 43. Shi, Q.H., et al., Rapid-Setting, Mesoporous, Bioactive Glass Cements that Induce Accelerated In Vitro Apatite Formation. Advanced Materials, 2006. 18(8): p. 1038-1042. 44. Sommerdijk, N.A.J.M. and G.d. With, Biomimetic CaCO3 Mineralization using Designer Molecules and Interfaces. Chemical Reviews, 2008. 108(11): p. 4499-4550. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10071 | - |
dc.description.abstract | 本研究探討了中空中孔洞氫氧基磷灰石奈米粒子 (hm-HANPs) 的製備及其在細胞內藥物傳輸的應用。在合成材料的部分,藉由調控乙二醇的多寡、形成碳酸鈣核的反應時間、醋酸鈣和碳酸氫鈉水溶液的濃度、磷酸水溶液的濃度與醋酸水溶液的多寡,探討對合成中空中孔洞氫氧基磷灰石奈米粒子 (hm-HANPs) 的影響。實驗結果顯示最佳合成條件如下: 碳酸鈣前趨液與乙二醇的體積比是1:5、形成碳酸鈣核的反應時間是3小時、碳酸鈣前趨液是 0.3 M 醋酸鈣與 0.3 M 碳酸氫鈉水溶液、0.01 M 磷酸水溶液以及碳酸鈣前趨液與醋酸水溶液的體積比須超過1:3.125。利用此最佳合成條件,可製備出粒俓均勻 (400 x 600 nm) 且單一分布的中空中孔洞氫氧基磷灰石奈米粒子。在細胞內藥物傳遞的部分,選擇抗癌藥doxorubicin (DOX)裝載於hm-HANPs的內部。實驗結果顯示,相較於實心結構的氫氧基磷灰石奈米粒子,中空中孔洞氫氧基磷灰石奈米粒子有較高 (i.e., 五倍) 的藥物負載量。此外,hm-HANPs的中孔洞殼減慢DOX從中空結構內部釋放出來的速度,進而減緩釋放初期的突釋現象。在不同的酸鹼值影響下,hm-HANPs也表現出不同的釋放行為。同時,與單獨使用DOX相較下,DOX-loaded hm-HANPs的抗癌療效有明顯的提升。可得知,利用這種DOX-loaded hm-HANPs易受酸鹼值影響的特性,將有助於其應用於可調控的藥物傳遞系統。 | zh_TW |
dc.description.abstract | This study reports the synthesis and intracellular drug delivery of hollow mesoporous hydroxyapatite nanoparticles (hm-HANPs). For the synthesis part, the effects of several critical factors including the amount of ethylene glycol (EG), reaction time to form CaCO3 cores, concentrations of Ca(CH3COO)2(aq) and NaHCO3(aq), concentration of H3PO4(aq) and the amount of CH3COOH(aq) were investigated for synthesizing hollow mesoporous hydroxyapatite nanoparticles (hm-HANPs). We optimized the reaction conditions as follows: The ratio of CaCO3 precursor solution to EG was 1:5. Reaction time to form CaCO3 cores was 3hr. The CaCO3 precursor solution was 0.3 M Ca(CH3COO)2(aq):0.3 M NaHCO3(aq). The concentrations of H3PO4(aq) was 0.01 M. And the ratio of the CaCO3 precursor solution to CH3COOH(aq) was over 1:3.125. The monodispersed hm-HANPs with a defined particle size (400 x 600 nm) could be achieved. For the intracellular drug delivery part, doxorubicin (DOX) was used as an anticancer drug and loaded into hm-HANPs. We found that hm-HANP exhibited higher (i.e., five times) drug loading capacity than HANPs without hollow core. In addition, mesoporous shell of hm-HANPs slowed down the release of loaded DOX, reducing the burst release at the beginning. Our hm-HANPs also exhibited pH-responsive release behavior. Compared with free DOX, the anticancer efficacy of DOX-loaded hm-HANPs was greatly enhanced. This pH-sensitive property of DOX-loaded, hm-HANPs would be useful for controlled drug delivery system. | en |
dc.description.provenance | Made available in DSpace on 2021-05-20T20:59:50Z (GMT). No. of bitstreams: 1 ntu-100-R98524017-1.pdf: 5963307 bytes, checksum: f9284fcf37b75a31662f7f4215ac2468 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | Table of Content
Abstract……………………………………………………………………………I 摘要………………………………………………………………………………II Table of Content……………………………………………………………………III List of Figure………………………………………………………………………V List of Table………………………………………………………………………VIII 1. Introduction…………………………………………………………………1 2. Literature review……………………………………………………………3 2.1 Drug carrier……………………………………………………………………3 2.1.1 Nanoscopic drug carriers…………………………………………………3 2.1.2 Comparison of organic and inorganic carriers…………………………4 2.2 Hydroxyapatite…………………………………………………………………6 2.2.1 Physicochemistry properties of hydroxyapatite (HA)…………………6 2.2.2 Synthesis of hollow hydroxyapatite nanoparticles………………………9 2.3 Anticancer drug……………………………………………………………12 2.3.1 Classification of anticancer drugs……………………………………12 2.3.2 The mechanism of doxorubicin (DOX)…………………………………17 3 Materials and experimental ………………………………………………19 3.1 Materials………………………………………………………………………19 3.2 Experimental apparatuses……………………………………………………20 3.3 Experimental methods………………………………………………………21 3.3.1 Synthesis of hollow mesoporous hydroxyapatite nanoparticles (hm-HANPs)……………………………………………………………21 3.3.2 Preparation of FITC-labeled hm-HANPs (FITC-hm-HANPs)………23 3.3.3 Preparation of DOX-loaded hm-HANPs or DOX-loaded FITC-hm-HANPs…………………………………………………………24 3.3.4 Measurement of release profiles of DOX from DOX-loaded hm-HANP or DOX-loaded HANP………………………………………………………25 3.3.5 Calculation for loading and release of doxorubicin………………………26 3.3.6 Cell culture…………………………………………………………………26 3.3.7 MTT assay………………………………………………………27 3.3.8 Preparation of samples for confocal fluorescence microscopy…………28 4 Results and discussion………………………………………………………29 4.1 Synthesis of hm-HANP………………………………………………………29 4.1.1 Effect of ethylene glycol on the morphology of CaCO3 cores……………29 4.1.2 Effect of reaction time on the morphology of CaCO3 cores……………31 4.1.3 Effect of the concentrations of the CaCO3 precursor on the morphology of CaCO3 cores………………………………………………………………32 4.1.4 Effect of the concentrations of H3PO4(aq) on the morphology of CaCO3/HA core/shell……………………………………………………………34 4.1.5 Effect of the amount of acetic acid aqueous solution on the removal of CaCO3 cores……………………………………………………………37 4.1.6 Properties of CaCO3 core, CaCO3/HA core/shell nanoparticles and hm-HANP synthesized in optimal conditions…………………………38 4.2 The application of hm-HANPs for intracellular drug delivery…………42 4.2.1 Loading of doxorubicin and their resulting release profiles…………42 4.2.1.1 Loading of doxorubicin…………………………………………………42 4.2.1.2 Release profiles of loaded DOX…………………………………………43 4.2.2 MTT assays and confocal images for BT-20 cell line…………………47 4.2.2.1 MTT assays……………………………………………………………47 4.2.2.2 Confocal images…………………………………………………………51 5 Conclusion……………………………………………………………………55 6 Reference………………………………………………………………………56 List of Figure Figure 2.1. SAEDs for the HAhexagonal, HAmonoclinic and OCP phase at different tilting angles: (a) 0 °, (b) +22 ° and (c) -22 °…………………………………………8 Figure 2.2. The family of anthracyclines…………………………………………18 Figure 3.1. Experimental flowchart for synthesis of hollow mesoporous hydroxyapatite nanoparticles (hm-HANPs)…………………………21 Figure 3.2. Experimental flowchart for preparation of FITC-labeled hm-HANP (FITC-hm-HANP)…………………………………………………23 Figure 3.3. Experimental flowchart for preparation of DOX-loaded hm-HANP or DOX-loaded FITC-hm-HANP…………………………………………24 Figure 3.4. Experimental flowchart for the measurement of release profiles of DOX from DOX-loadd hm-HANP……………………………………25 Figure 3.5. Experimental flowchart of MTT assay…………………………………………………………………………27 Figure 3.6. Experimental flowchart for preparation of samples for confocal fluorescence microscope…………………………………………………28 Figure 4.1. SEM images of CaCO3 cores using different ratios of the CaCO3 precursor solution to ethylene glycol. (a) 1:0, (b) 1:1, (c) 1:3, (d) 1:5, (e) 1:8, and (f) 1:10.………………………………………………………30 Figure 4.2. SEM images of CaCO3 cores synthesized for different reaction time. (a) 3 hr, (b) 12 hr, and (c) 24 hr………………………………………32 Figure 4.3. SEM images of CaCO3 cores using different concentrations and ratios of Ca(CH3COO)2(aq) and NaHCO3(aq). (a) 0.3 M:0.3 M, (b) 0.3 M:1.0 M, (c) 0.5 M:0.5 M, and (d) 1.0 M:1.0 M……………………………33 Figure 4.4. SEM images of CaCO3/HA core/shell nanoparticles using different concentrations of H3PO4(aq). (a) 0.3 M, (b)0.1M, (c) 0.05 M, (d) 0.01 M, and (e) 0.0025 M………………………………………………………35 Figure 4.5. TEM images of (a) CaCO3 cores and (b) CaCO3/HA core/shell nanoparticles……………………………………………………………36 Figure 4.6. TEM images of CaCO3/HA core/shell nanoparticles, where CaCO3 cores were removed by using different ratios of CaCO3 precursor solution to CH3COOH(aq). (a) 1:2.5, (b) 1:2.75, (c) 1:3, (d) 1:3.125, and (e) 1:3.25………………………………………………………………37 Figure 4.7. SEM images of (a) CaCO3 cores, (b) CaCO3/HA core/shell nanoparticles and (c) hm-HANPs in optimal conditions; TEM images of (d) CaCO3 cores, (e) CaCO3/HA core/shell nanoparticles and (f) hm-HANPs in the optimal conditions………………………………………………………………40 Figure 4.8. XPD patterns of (a) CaCO3 core and (b) hm-HANP in the optimal conditions………………………………………………………………41 Figure 4.9. (a) N2 adsorption/desorption isotherm and (b) corresponding pore size distribution of hm-HANP in the optimal conditions……………41 Figure 4.10. (a) Released amount and (b) released percentage of DOX from DOX-loaded hm-HANPs loaded in different concentration of …………………………………………………………………………45 Figure 4.11. (a) Released amount and (b) released percentage of DOX from DOX-loaded HANPs and DOX-loaded hm-HANPs………………46 Figure 4.12. Released percentage of DOX from DOX-loaded hm-HANPs in different pH-value PBS solutions……………………………………46 Figure 4.13. MTT assays of hm-HANP and DOX-loaded hm-HANP incubated with BT-20 cells for 24 hr……………………………………………48 Figure 4.14. MTT assays of (a) free DOX and (b) DOX-loaded hm-HANP incubated with BT-20 cells for 24 hr………………………………49 Figure 4.15. MTT assays of DOX-loaded hm-HANP incubated with BT-20 cells for 4, 8, 12 and 24 hr…………………………………………………50 Figure 4.16. Confocal images of BT-20 cells incubated with FITC-hm-HANPs (50 μg/mL) for 24 hr…………………………………………………52 Figure 4.17. Confocal images of BT-20 cells incubated with FITC-hm-HANPs (100 μg/mL) for 24 hr…………………………………………………54 Figure 4.18. Confocal images of BT-20 cells incubated with DOX-loaded FITC-hm-HANPs (100 μg/mL) for 24 hr……………………………57 List of Table Table 2.1 Some of calcium phosphate phases obtained from liquid precipitation ……………………………………………6 Table 2.2. Anticancer drugs directed against tumor DNA……………………13 Table 2.3. Anticancer drugs directed against tumor RNA……………………14 Table 2.4. Anticancer drugs directed against proteins in the tumor…………15 Table 2.5. Anticancer drugs directed against the endothelium…………………16 Table 2.6. Anticancer drugs directed against the extracellular matrix…………6 Table 2.7. Anticancer drugs directed against the host cells…………………16 Table 3.1. All chemicals used in this study………………………………………19 Table 4.1. Loaded capacity and loaded efficiency of DOX-loaded hm-HANP in different concentrations of DOX(aq)………………………………………………42 Table 4.2. Basic properties, loaded capacity and loaded efficiency of different materials…………………………………………………………………………43 Table 4.3. The kinetics and related parameters of release of loaded DOX in different conditions………………………………………………………………45 Table 4.4. The half maximal inhibitory concentration (IC50) of hm-HANPs, DOX-loaded hm-HANPs and free DOX incubated for different times in the BT-20 cell line……………………………………………………………………50 | |
dc.language.iso | en | |
dc.title | 中空中孔洞氫氧基磷灰石奈米粒子的製備及細胞內藥物傳輸的應用 | zh_TW |
dc.title | Synthesis of Hollow Mesoporous Hydroxyapatite Nanoparticles for Intracellular Drug Delivery | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林?輝(Feng-Huei Lin),徐振哲(Cheng-Che(Jerry),陳林祈(Lin-Chi Chen),楊家銘(Chia-Min Yang) | |
dc.subject.keyword | 氫氧基磷灰石,中空,奈米粒子,doxorubicin,BT-20癌細胞,藥物載體,藥物傳遞, | zh_TW |
dc.subject.keyword | hydroxyapatite,hollow,nanoparticles,doxorubicin,BT-20 cancer cells,drug carrier,drug delivery, | en |
dc.relation.page | 59 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2011-07-25 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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