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  1. NTU Theses and Dissertations Repository
  2. 工學院
  3. 醫學工程學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17326
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor林?輝
dc.contributor.authorWan-Tzu Loen
dc.contributor.author羅婉慈zh_TW
dc.date.accessioned2021-06-08T00:06:57Z-
dc.date.copyright2013-08-20
dc.date.issued2013
dc.date.submitted2013-08-12
dc.identifier.citation1. Hall, J.E. and A.C. Guyton, Guyton and Hall textbook of medical physiology. 2011, Philadelphia, PA: Saunders/Elsevier.
2. Gardella, T.J. and H.M. Kronenberg, Parathyroid Hormone (PTH), in Encyclopedia of Endocrine Diseases, M. Editor-in-Chief: Luciano, Editor. 2004, Elsevier: New York. p. 513-520.
3. La Perle, K.M.D. and C.D. Jordan, 15 - Endocrine System, in Comparative Anatomy and Histology, M.T. Piper, et al., Editors. 2012, Academic Press: San Diego. p. 211-227.
4. Marieb, E.N. and K. Hoehn, Human anatomy & physiology. 2007, San Francisco, Calif.: Pearson Benjamin Cummings.
5. Shoback, D., Hypoparathyroidism. New England Journal of Medicine, 2008. 359(4): p. 391-403.
6. Testini, M., et al., Hypoparathyroidism after total thyroidectomy. Minerva Chirurgica, 2007. 62(5): p. 409-15.
7. Hasse, C., et al., First successful xenotransplantation of microencapsulated human parathyroid tissue in experimental hypopara-thyroidism: Long-term function without immunosuppression. Journal of Microencapsulation, 1997. 14(5): p. 617-626.
8. Chang, T.M., Semipermeable Microcapsules. Science, 1964. 146(3643): p. 524-5.
9. Zhang, Y., et al., 5.09 - The Artificial Organ: Cell Encapsulation, in Comprehensive Biotechnology (Second Edition), M.-Y. Editor-in-Chief: Murray, Editor. 2011, Academic Press: Burlington. p. 99-114.
10. Sundararaghavan, H.G. and J.A. Burdick, 5.509 - Cell Encapsulation, in Comprehensive Biomaterials, D. Editor-in-Chief: Paul, Editor. 2011, Elsevier: Oxford. p. 115-130.
11. Nafea, E.H., P.-W.A.M. L.A, and P.J. Martens, Immunoisolating semi-permeable membranes for cell encapsulation: Focus on hydrogels. Journal of Controlled Release, 2011. 154(2): p. 110-122.
12. Chang Oc, M.D.C.M.P.F.T.M.S. and S. Prakash PhD, Therapeutic uses of microencapsulated genetically engineered cells. Molecular Medicine Today, 1998. 4(5): p. 221-227.
13. Hasse, C., et al., Parathyroid Xenotransplantation without Immunosuppression in Experimental Hypoparathyroidism: Long-term In Vivo Function following Microencapsulation with a Clinically Suitable Alginate. World Journal of Surgery, 2000. 24(11): p. 1361-1366.
14. Barbosa, M.A., A.P. Pego, and I.F. Amaral, 2.213 - Chitosan, in Comprehensive Biomaterials, D. Editor-in-Chief: Paul, Editor. 2011, Elsevier: Oxford. p. 221-237.
15. Bhattarai, N., J. Gunn, and M. Zhang, Chitosan-based hydrogels for controlled, localized drug delivery. Advanced Drug Delivery Reviews, 2010. 62(1): p. 83-99.
16. Ko, J.A., et al., Preparation and characterization of chitosan microparticles intended for controlled drug delivery. International Journal of Pharmaceutics, 2002. 249(1–2): p. 165-174.
17. Krohn, R.I., The Colorimetric Detection and Quantitation of Total Protein, in Current Protocols in Cell Biology. 2011, John Wiley & Sons, Inc.
18. Mosmann, T., Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 1983. 65(1–2): p. 55-63.
19. Moskalenko, V., et al., Preoperative evaluation of microencapsulated human parathyroid tissue aids selection of the optimal bioartificial graft for human parathyroid allotransplantation. Transplant International, 2007. 20(8): p. 688-696.
20. Zhang, C., et al., Novel Chitosan-Derived Nanomaterials and Their Micelle-Forming Properties. Journal of Agricultural and Food Chemistry, 2006. 54(22): p. 8409-8416.
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17326-
dc.description.abstract副甲狀腺機能不足症為目前難以完全治癒的內分泌疾病之一,在臨床上也並不建議在副甲狀腺異體移植後使用全身性免疫抑制的藥物,故使用具半通透性的膜來包覆副甲狀腺組織再進行異體移植,來達到植入組織在病人體內具有免疫隔離的效果。本研究使用氧化幾丁聚醣交聯昆布胺酸,將副甲狀腺組織塊外面包覆後滴入三聚磷酸鈉溶液中,利用離子吸附的方式在組織塊外圍成一層膜狀的水膠。在材料測試方面,氧化幾丁聚醣/昆布胺酸水膠在體外28天內可維持約80 %重量與型態,且具有良好的生物相容性,同時不具細胞毒性。以材料包覆組織後進行體外培養結果,組織外的膜狀水膠可使人類副甲狀腺素成功釋放至環境中,證實包覆材料並不會影響組織的分泌功能,並大致具有隨周圍環境的鈣離子濃度調整釋放量的趨勢,且在組織切片的觀察結果顯示包覆材料並不影響具分泌能力的主細胞的存活。在動物實驗方面雖有成功導致實驗動物具有副甲狀腺機能不足症,但在移植之後的組織對血鈣濃度的調節效果並不如預期,顯示本實驗模型還有待改進。zh_TW
dc.description.abstractPermanent hypoparathyroidism is one of the endocrine disorders that are difficult to treat medically. Since hypoparathyroidism rarely is a life-threatening condition, systemic immunosuppression for recipients of allo-transplants are not justified. Coating the allo-transplanted tissue with a semipermeable membrane to protect the tissue from immune rejection is an alternative. In this study, oxidized chitosan cross-linked with laminin is prepared for microencapsulation of parathyroid tissue fragments. The parathyroid tissue particles with oxidized chitosan/laminin are dropped in sodium tripolyphophate solution. Tripolyphophate ion, the polyanion, would interact electrostatically with the cationic chitosan and become hydrogel to enclose the tissue particle. In the material analysis, oxidized chitosan/laminin hydrogel maintained about 80 % mass and it’s morphology in PBS until the 28th day. The material in vitro tests also showed that hydrogel had good biocompatibility and low cytotoxicity. The microencapsulation of parathyroid tissue in vitro study revealed that the hydrogel was freely permeable to the human parathyroid hormone and responded to different calcium concentration in the ex vivo test. The histological evaluation showed that the capsular hydrogel membrane were not affective to the chief cells which are the main secretory cell in the parathyroid glands. In animal study, hypocalcemia rats were induced successfully but the encapsulated tissue particles implanted were not functional in vivo. The result demonstrated that the animal model in this study must be improved.en
dc.description.provenanceMade available in DSpace on 2021-06-08T00:06:57Z (GMT). No. of bitstreams: 1
ntu-102-R99548039-1.pdf: 2624647 bytes, checksum: ef8a0c25375263479af950857a12fe87 (MD5)
Previous issue date: 2013
en
dc.description.tableofcontents口試委員審定書 i
誌謝 ii
中文摘要 iii
英文摘要 iv
目錄 v
圖目錄 ix
表目錄 xii
第一章 簡介 1
1.1 副甲狀腺(parathyroid) 1
1.2 副甲狀腺機能不足症(hypoparathyriodism) 3
1.3 副甲狀腺異體移植(parathyroid allotransplant) 4
1.4 免疫隔離(immunoisolation) 5
1.5 研究目的 8
第二章 文獻回顧 9
2.1 目前用於副甲狀腺異體移植免疫隔離的研究 9
2.2 幾丁聚醣水膠(chitosan-based hydrogel) 13
第三章 材料與實驗方法 15
3.1 實驗儀器與實驗藥品 15
3.1.1 實驗儀器 15
3.1.2 實驗藥品 15
3.2 氧化幾丁聚醣的製備方法與其特性 17
3.2.1 氧化幾丁聚醣的製備方法 17
3.2.2 特性官能基測定 17
3.2.3 核磁共振光譜儀分析 17
3.3 氧化幾丁聚醣/昆布胺酸水膠製備方法與其特性 18
3.3.1 氧化幾丁聚醣/昆布胺酸水膠製備方法 18
3.3.2 In vitro降解程度測定 18
3.3.3 切割分子量(molecular weight cut-off, MWCO)測定 18
3.4 氧化幾丁聚醣/昆布胺酸水膠細胞活性與細胞毒性測試 20
3.4.1 細胞存活/增生檢測 20
3.4.2 細胞毒性測試 21
3.5 氧化幾丁聚醣/昆布胺酸水膠包覆副甲狀腺組織 23
3.5.1 副甲狀腺組織in vitro培養與包覆 23
3.5.2 副甲狀腺組織in vitro分泌能力與ex vivo周流系統試驗 23
3.5.3 In vitro培養組織塊切片及染色 25
3.6 動物實驗 26
3.6.1 副甲狀腺機能不足動物模型建立 26
3.6.2 動物實驗分組 28
3.6.3 血液生化檢測 28
第四章 結果與討論 29
4.1 氧化幾丁聚醣性質分析 29
4.1.1 紅外線光譜儀分析 29
4.1.2 核磁共振光譜儀分析 31
4.2 氧化幾丁聚醣/昆布胺酸水膠特性分析 32
4.2.1 In vitro降解程度測定 32
4.2.2 切割分子量測定 34
4.3 氧化幾丁聚醣/昆布胺酸水膠細胞活性與細胞毒性測試 36
4.3.1 細胞存活/增生檢測 36
4.3.2 細胞毒性測試 37
4.4 副甲狀腺組織in vitro分泌能力與ex vivo周流系統試驗結果 38
4.4.1 副甲狀腺組織in vitro分泌能力 38
4.4.2 副甲狀腺組織ex vivo周流系統試驗結果 39
4.4.3 In vitro培養組織塊切片及染色 40
4.5 動物實驗 43
第五章 結論 45
參考資料 46
dc.language.isozh-TW
dc.title氧化幾丁聚醣/昆布胺酸水膠作為異體副甲狀腺移植免疫隔離之運用zh_TW
dc.titleOxidized Chitosan/Laminin Hydrogel for Allo-parathyroid Immunoisolationen
dc.typeThesis
dc.date.schoolyear101-2
dc.description.degree碩士
dc.contributor.oralexamcommittee張國基,林建成,郭士民,楊凱強
dc.subject.keyword副甲狀腺,副甲狀腺機能不足,異體移植,免疫隔離,幾丁聚醣水膠,微包覆,zh_TW
dc.subject.keywordparathyroid glands,hypoparathyroidism,allo-transplantation,immunoisolation,chitosan-based hydrogel,microencapsulation,en
dc.relation.page47
dc.rights.note未授權
dc.date.accepted2013-08-13
dc.contributor.author-college工學院zh_TW
dc.contributor.author-dept醫學工程學研究所zh_TW
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