臺灣博碩士論文加值系統

中空纖維可被利用作為一種可調控的釋放裝置（control release device），如藥物包埋或細胞包埋的載體。一般用的中空纖維材質為再生纖維素（Regenerated Cellulose），是以植物纖維純化出來的纖維素（cellulose）製造，其分子結構為多醣體（polysaccharide），表面有氫氧基的存在，因此植入人體時，易引發體內免疫系統，而將中空纖維透析膜當成感染源，活化補體反應。所以當中空纖維的生物載體植入體內，將會造成一連串的生物反應，最後在此載體外部產生纖維化，造成植入物的功能減退甚至喪失效果。
聚乙二醇（polyethylene glycol，PEG）為長鏈親水性高分子，沒有分枝與多餘的不飽和鍵，無法形成穩固的網狀結構，所以不能作為包覆藥物的載體；不過其熱穩定性高，生物相容性高，具有生物不可分解、不易水解與低毒性等多種特性。如果材質表面有聚乙二醇的存在，可以減少蛋白質的吸附，降低凝血反應和纖維母細胞的吸附，因而聚乙二醇常常被應用在修飾材料表面以及改變標的物性質的高分子。
本論文利用化學與電漿合成方式，以聚乙二醇修飾中空纖維的薄膜表面，並由ESCA與螢光分析顯示兩種方式確認。經由通透性質測試，聚乙二醇修飾的中空纖維具有較佳的選擇性。進一步以動物實驗分析，發現修飾後的中空纖維的確可以減少細胞吸附及纖維化的產生。將修飾後的中空纖維應用於副甲狀腺細胞包覆實驗，在體外及體內測試中，副甲狀腺細胞皆維持良好的功能，期望將來可利用於複合生物人工臟器（artificial organ），以解決副甲狀腺功能減退的問題。

Hollow fiber could be used to fabricate devices for controlled release of bioactive entities such as cells and drugs. Of the many materials of hollow fiber, regenerated cellulose is probably the most common one. When hollow fiber is implanted in the tissue, it usually induces a series of tissue reactions leading to fibrosis around the hollow fiber and finally resulting in the loss of the designed functions.
Long chain polyethylene glycol (PEG) is a thermal stable, hydrophilic polymer with excellent biocompatibility and low toxicity. When the surface of a material is coated with PEG, a great reduction of protein absorption, blood clotting and fibroblast attachment can be achieved. This results in a great reduction of fibrosis on the surface of implant. Therefore, PEG is often used to modify the surface of material for various biomedical applications.
Using chemical and plasma modifications, PEG can be coated onto the surface of cellulose hollow fiber, which results in increasing permeated selectivity. Animal implant test results indicate that surface modification with PEG reduces fibrosis phenomenon. The modified hollow fiber is analyzed and investigated as the implant device for parathyroid xenograft. Parathyroid cells encapsulated with modified cellulose hollow fiber still maintain their physiological function in vitro and in vivo. The technique of using this hollow fiber to immobilize parathyroid cells provide a feasible clinical therapy for parathyroid malfunction.

謝誌 I
中文摘要 II
Abstract III
目錄 IV
圖表索引 VIII
第一章：緒論 1
1-1中空纖維薄膜 1
1-1-1透析原理 3
1-1-2薄膜的材質 5
1-1-2-1纖維素薄膜 5
1-1-2-2非纖維素薄膜 7
1-1-3薄膜修飾 8
1-1-3-1化學修飾 8
1-1-3-2電漿修飾 9
1-1-4薄膜消毒 9
1-1-4-1高溫消毒 10
1-1-4-2蒸氣消毒 10
1-1-4-3環氧乙烯氣消毒 10
1-1-4-4輻射線照射 11
1-1-5薄膜之醫工應用 12
1-1-5-1血液透析 12
1-1-5-2人工臟器 15
1-1-5-3人工胰臟 15
1-1-5-4人工肝臟 18
1-1-6結語 20
1-2聚乙二醇 22
1-2-1分類 22
1-2-2物性和化性 23
1-2-3生物特性 24
1-2-4毒性 25
1-2-5改質 26
1-2-6應用 28
1-3副甲狀腺 29
1-3-1副甲狀腺細胞 29
1-3-2副甲狀腺荷爾蒙 30
1-3-2-1 PTH在腎臟的作用 31
1-3-2-2 PTH在骨骼的作用 31
1-3-3副甲狀腺疾病 33
1-4實驗動機與目的 35
第二章：實驗藥品、材料及設備 37
2-1實驗藥品與材料 37
2-2實驗設備儀器與操作方法 40
第三章：實驗方法 42
3-1中空纖維之表面修飾 42
3-1-1電漿方式修飾 42
3-1-1-1合成螢光的聚乙二醇（MPEG-FITC） 42
3-1-1-2凝膠管柱層析分離 43
3-1-1-3電漿方式製程 44
3-1-2化學方式修飾 45
3-1-2-1合成CM-MPEG（carboxymethyl- methoxypolyethylene glycol） 45
3-1-2-2化學方式製程 46
3-1-3中空纖維表面聚乙二醇之定量 47
3-1-3-1電漿方式修飾之螢光鑑定與分析-酸洗出法 47
3-1-3-2電漿方式修飾之螢光鑑定與分析-Densitometer scan 47
3-1-3-3表面元素分析（ESCA） 48
3-1-4中空纖維的分子量劃分（MWCO）測試 48
3-2中空纖維之生物相容性測試 51
3-2-1修飾後中空纖維之動物實驗 51
3-2-1-1動物皮下植入 52
3-2-1-2組織包埋與染色 52
3-2-2灌注聚乙二醇之修飾後中空纖維的動物實驗 55
3-2-2-1聚乙二醇之細胞毒性測試 55
3-2-2-2動物皮下植入 56
3-3副甲狀腺之研究 57
3-3-1副甲狀腺細胞之分離 57
3-3-2副甲狀腺細胞與聚乙二醇之毒性測試 58
3-3-3以中空纖維包覆副甲狀腺細胞 59
3-3-4體外副甲狀腺荷爾蒙釋放測定 59
3-4人工副甲狀腺體之駐植 59
3-4-1動物處理 59
3-4-2人工副甲狀腺體植入動物體內 60
3-4-3血鈣之測定 60
第四章：結果與討論 62
4-1中空纖維表面改質 62
4-1-1合成螢光的聚乙二醇（MPEG-FITC） 62
4-1-2凝膠管柱層析分離 63
4-1-3電漿方式修飾之中空纖維 66
4-1-3-1電漿方式修飾之螢光鑑定與分析-酸洗出法 66
4-1-3-2電漿方式修飾之螢光鑑定與分析-Densitometer scan 67
4-1-3-3電漿方式修飾的表面元素分析（ESCA） 70
4-1-3-4化學方式修飾的表面元素分析（ESCA） 77
4-1-4中空纖維的劃分分子量測試 81
4-2中空纖維表面之生物相容性測試 89
4-2-1修飾後中空纖維之動物實驗 89
4-2-2聚乙二醇之細胞毒性測試 99
4-2-3灌注聚乙二醇之中空纖維皮下植入 100
4-3副甲狀腺之研究 111
4-3-1副甲狀腺細胞之型態 111
4-3-2副甲狀腺細胞與聚乙二醇之毒性測試 113
4-3-3副甲狀腺荷爾蒙釋放測定 114
4-3-4人工副甲狀腺體駐植 114
第五章 結論 120
第六章：參考資料 122

1. Prehn, RT., Weaver, JM., and Algire, GH. (1954) J. National Cancer Institute : V. 15, n3, 509-517.
2. Lee JH. Kopecek J. Andrade JD. "Protein-resistant surfaces prepared by PEO-containing block copolymer surfactants." (1989) Journal of Biomedical Materials Research : 23, 351-68.
3. Desai NP. Hubbell JA. "Biological responses to polyethylene oxide modified polyethylene terephthalate surfaces." (1991) Journal of Biomedical Materials Research. : 25, 829-43.
4. Sawhney AS. Hubbell JA. "Poly(ethylene oxide)-graft-poly(L-lysine) copolymers to enhance the biocompatibility of poly(L-lysine)-alginate microcapsule membranes." (1992) Biomaterials. 13(12) : 863-70.
5. Kishida A. Mishima K. Corretge E. Konishi H. Ikada Y. "Interactions of poly(ethylene glycol)-grafted cellulose membranes with proteins and platelets." (1992) Biomaterials : 13, 113-8.
6. Sawhney AS., Pathak CP. and Hubbell JA. "Interfacial photo-polymerization of poly(ethylene glycol)-based hydrogels upon alginate-poly(l-lysine) microcapsules for enhanced biocompatibility." ( 1993) Biomaterials : 14(13), 1008-1016.
7. Chen BS. Chen GC. Hsieh JH. Meng MH. Tsai TF. Liu JJ. Huang JH. Chang TH. "Large-scale production of monoclonal antibodies using hollow fiber bioreactor." (1990) Chinese Journal of Microbiology & Immunology. 23(1):75-88.
8. Clark K. and Colton, "Engineering challenges in cellencapsulation technology." (1996) Trends In Biotechnology : 14, 158-62.
9. Morris PJ. "Immunoprotection of therapeutic cell transplants by encapsulation". (1996) Trends In Biotechnology : 14, 163-167.
10.黃志強，血液透析學，合記圖書出版社，1997。
11.洪聰民、洪石獅、李昭仁、嚴燦鑫、陳劉旺， "高分子薄膜與血液淨化" (1987) 化工, 136, 55-92.
12.Landfield H, "Sterilization of medical devices based on polymer selection and stabilization technique" in Biocompatible polymers, metals and composition, edited by Szycher M., (1983) Ch. 43, Techonomic publishing.
13.Asanuma, Akira, Taneda, Nobuo, Hasegawa, Ryozo, Jindai, Shohei, Morishita, Toshio. US Patent 4176156, "Method for heat-sterilizing artificial kidneys." Teijin Limited, Japan : 1979.
14.Royer GP. and Anantharmaiah GM. (1979) J. Am. Chem. Soc. : 101, 3394-3396.
15.穗積啟一郎（編）超LSI時代之Plasma Chemistry，化學工業調查會(1983).
16.長田義人（編）Plasma重和，東京化學問人(1986).
17.M. Gazicki, et al, (1986) Journal of Polymer Science. Polymer Chemistry Edition. : 24, 215.
18.Ward RS., White KA., Wolcott CA., Wang AY., Kuhn RW., Taylor JE. and John JK., "Development of a hybrid artificial pancreas with a dense polyurethane membrane." (1993) ASAIO Journal. 39(3):M 261-267.
19.Chick WL. Like AA. Lauris V. "Beta cell culture on synthetic capillaries: an artificial endocrine pancreas." (1975) Science : 187(4179), 847-849.
20.Chick WL., Perna JJ., Lauris V., Low D., Galletti PM., Panol G., Whittemore AD., Like AA., Colton CK. and Lysaght MJ., "Artificial pancreas using living beta cells: effects on glucose homeostasis in diabetic rats." (1977) Science : 197(4305), 780-782.
21.Gates RJ. and Lazarus NR. "Reversal of streptozotocin-induced diabetes in rats by intraperitoneal implantation of encapsulated neonatal rabbit pancreatic tissue." (1977) Lancet : II, 1257-1259.
22.Valente U., Ferro M., Campisi C., Parodi F. and Tosatti E., "Allogeneic pancreatic islet transplantation by means of artificial membrane chambers in 13 diabetic recipients." (1980) Transplantation Proceedings : 12(4 Suppl 2):223-226.
23.Reach G., Jaffrin MY. and Desjeux JF. "A U-shaped bioartificial pancreas with rapid glucose-insulin kinetics. In vitro evaluation and kinetic modelling." (1984) Diabetes : 33, 752-761.
24.Lim F. and Sun AM. ". Microencapsulated islets as bioartificial endocrine pancreas." (1980) Science : 210, 908-910.
25.O'Shea GM., and Sun AM. "Encapsulation of rat islets of Langerhans prolongs xenograft survival in diabetic mice." (1986) Diabetes : 35, 943-946.
26.Altman JJ., Houlbert D. and Callard P. "Long-term plasma glucose normalization in experimental diabetic rats with macroencapsulated implants of benign human insulinomas." (1986) Diabetes : 35, 625-633.
27.Tsuruta T., Hayashi T., Kataoka K., Inhihara K. and Kimura Y. (1993) Biomedical Application of Polymeric Materials. CRC Press.
28.Paul E. and Lacy, (1995) Scientific American : July, 50-58.
29.Mikos AG., Papadaki MG., Kouvroukoglou S., Ishaug SL. and Thomson RC. "Mini-Review: Islet transplantation to create a bioartificial pancreas." (1994) Biotechnology and Bioengineering : 43, 673-677.
30.Sarver JG. and Fournier RL., "Numerical investigation of a novel spiral wound membrane sandwich design for an implantable bioartificial pancreas." (1990) Computers in Biology & Medicine : 20, 105-119.
31.Horcher A., Zekorn T., Siebers U., Klock G., Schnettler R., Arnold M., Federlin K., Zimmermann U. and Bretzel RG., "Insulin release from different models of a bioartificial pancreas (microencapsulation versus alginate-coating)." (1992) Transplantation Proceedings. 24(6), 2950-2951.
32.Soon-Shiong P., Heintz R., Yao Z., Yao Q., Sanford P., Lanza RP. and Meredith N., "Glucose-insulin kinetics of the extravascular bioartificial pancreas. A study using microencapsulated rat islets." (1992) ASAIO J. : 38, 851-854.
33.Lanza RP., Sullivan SJ. and Chick WL., ". Perspectives in diabetes. Islet transplantation with immunoisolation." (1992) Diabetes : 41, 1503-1510.
34.Chick WL., Perna JJ. and Lauris V., "Artificial pancreas using living beta cells:. effects on glucose homeostasis in diabetic rats." (1977) Science: 197, 780-782.
35.Sun AM., Parisiun W., Healy GM. and Vacek I. "The use, in diabetic rats and monkeys, of artificial capillary units containing cultured islets of Langerhans (artificial endocrine pancreas)." (1977) Diabetes : 26, 1136-1139.
36.Tze WJ., Wong FC. and Chen LM. "Implantable artificial capillary unit for pancreatic islet allograft and xenograft. Diabetologia." (1979) Diabetologia : 16, 247-252.
37.Reach G., Poussier P., Sausse A., Assan R., Ltoh M. and Gerich J.E. "Functional evaluation of a bioartificial pancreas using isolated islets perifused with blood ultrafiltrate." (1981) Diabetes : 30, 296-301.
38.
39.Altman JJ., MacMillan P., Callard P. and Galletti PM. (1986) "A bioartificial pancreas prevents chronic complications of diabetes in rats." Trans. Am. Soc. Artif. Intern. Organs. : 32, 145-147.
40.Fan MY., Lum ZP., Fu XW., Levesque L., Isabella TT. and Anthony MS. "Reversal of diabetes in BB rats by transplantation of encapsulated pancreatic islets." (1990) Diabetes : 39, 519-522.
41.Kessler L., Pinget M., Aprahamian M., Dejardin P. and Damge C. ". In vitro and in vivo studies of the properties of an artificial membrane for pancreatic islet encapsulation." (1991) Horm. Metab. Res. : 23, 312-317.
42.Lacy PE., Hegre OD., Gerasimidi-Vazeou A., Gentile FT. and Dionne KE. "Maintenance of normoglycemia in diabetic mice by subcutaneous xenografts of encapsulated islets." (1991) Science : 254, 1782-1784.
43.Inoue K., Fujisato T., Gu YJ., Burczak K., Sumi S., Kogire M., Tobe T., Uchida K., Nakai I., Maetani S. and Ikada Y. "Experimental hybrid islet transplantation: application of polyvinyl alcohol membrane for entrapment of islets." (1992) Pancreas : 7, 562-568.
44.Lanza RP., Kuhtreiber WM., Ecker D., Staruk JE., Chick WL., "Xenotransplantation of porcine and bovine islets without immunosuppression using uncoated alginate microspheres." (1995) Transplantation : 59, 1377-1384.
45.Tze WJ., Tai J., Wong FC. and Davis HR. "Studies with implantable artificial capillary units containing rat islets on diabetic dogs." (1980) Diabetologia : 19, 541-545.
46.Maki T., Ubhi CS., Sanchez-Farpon H., Sultan SJ., Borland K., Muller TE., Solomon BA., Chick WL., and Monaco AP. "Successful treatment of diabetes with the biohybrid artificial pancreas in dogs." (1991) Transplantation : 51, 43-51.
47.Sun AM., Cai Z., Shi Z., Ma F. and O'Shea GM. (1986) Trans. Am. Soc. Artif. Intern. Organs. : 32, 39-41.
48.Jauregui HO., Bengochea L., Naik S., and Driscoll JL. "P-450 activity in monolayer cultures of hepatocytes seeded in polystyrene tissue culture dishes." (1986) Trans. Am. Soc. Artif. Intern. Organs. : 32, 357-359.
49.Miura Y., Akimoto T, Kanazawa H. and Yagi K. "Synthesis and secretion of protein by hepatocytes entrapped within calcium alginate." (1986) Artif. Organs. : 10, 460-465.
50.Cai Z., Shi Z., O'Shea G., and Sun AM. (1988) "Microencapsulated hepatocytes for bioartificial liver support." Artif. Organs. : 12, 388-393.
51.Takahama T., Yamazaki Z., Iizuka I., Kanai F., Hiraishi M., Wada T., Fujimori Y., Asano K., Sonoda T., "Prototype of hybrid artificial liver assist system." (1983) Transactions - American Society for Artificial Internal Organs. 29, 475-479.
52.Wong H., and Chang TM. "Bioartificial liver: implanted artificial cells microencapsulated living hepatocytes increases survival of liver failure rats." (1986) Int. J. Artif. Organs. : 9, 335-336.
53.Saito S., Sakagami K. and Orita K. "A new hybrid artificial liver using a combination of hepatocytes and biomatrix." (1987) ASAIO Transactions. 33(3):459-62.
54.Uchino J., Tsuburaya T., Kumagai F., Hase T., Hamada T., Komai T., Funatsu A., Hashimura E., Nakamura K. and Kon T., "A hybrid bioartificial liver composed of multiplated hepatocyte monolayers." (1988) ASAIO Transactions. 34(4), 972-7.
55.Yang Y., Jacobs H., Bae YH. and Kim SW. (1994) Biotechnology and Bioengineering : 43, 207-214.
56.Saksena S. and Zydney AL. (1994) Biotechnology and Bioengineering : 43, 960-968.
57.Li RH. and Barbari TA. (1994) Journal Membrane Sci. : 88, 115-125.
58.Michaels As. Hight flow membrane. US Patent 361502.
59.黃榮茂、王禹文、林聖富、楊得仁編譯(1989)化學化工百科辭典，第二版，曉園出版社 : 762頁。
60.Budavari S., Maryadele J., O'Neil AS., Patricia E., Heckelman J. and Kinneary, F., (1997) The Merck Index, Twelfth Edition, An Encyclopedia of Chemicals, Drugs, and Biologicals.
61.Mori Y., Nagaoka S., Takiuchi H., Kikuchi T., Noguchi N., Tanzawa H. and Noishiki Y., "A new antithrombogenic material with long polyethyleneoxide chains." (1982) Transactions - American Society for Artificial Internal Organs. 28, 459-63.
62.Woodle MC. and Lasic DD., "Sterically stabilized liposomes." (1992) Biochimica et Biophysica Acta. 1113(2), 171-99.
63.王奕嵐, (1997) "透明質酸還原端氧化衍生物之應用", 國立陽明大學醫工所碩士論文.
64.許峻源, (1998) "短鏈褐藻酸還原端接枝甲氧基聚乙二醇之團鏈共聚物的製備與定性", 國立陽明大學醫工所碩士論文.
65."Topical PEG in Burn Ointments", (1982) FAD Drug Bull : 12, 25-36.
66.P. R. Wheater, H. G. Burkitt, and V. G. Daniel, (1987) Functional Histology, 2thed.
67.Wu HL., (1995) Journal of the China Textile Institute : 5, 79-83.
68.Editor-in-chief, John P. Bilezikian ; associate editors, Robert Marcus, Michael A. Levine, (1993) The Parathyroids : basic and clinical concepts, New York, Raven Press.
69. Eknoyan G. "A history of the parathyroid glands." (1995), American Journal of Kidney Disease, 26(5), 801-807.
70.林毓萌、張慶忠、陳芳武， "副甲狀腺素" (1984) 當代醫學，11(11), 29-36.
71.Mayer GP. and Hurst JG., "Sigmoidal relationship between para-thyroid hormone secretion rate and plasma calcium concentration in calves." (1978), Endocrinology, 102, 1036-1042.
72.Bruno Niederle, Rudolf Roka, and Murray F B., "The transplantation of parathyroid tissue in man: development, indications, technique, and results." (1982), Endocrine Reviews, 3(3), 245-279.
73. Hamming JF., Van de Velde CJ., Goslings BM., Schelfhout LJ., Fleuren GJ., Hermans J. and Zwaveling A. "Prognosis and morbidity after total thyroidectomy for papillary, follicular and medullary thyroid cancer." (1989) European Journal of Cancer Clin. and Oncology, 25, 1317-1324.
74.Hasse C., Schrezenmeir J., Stinner B., Schark C., Wagner P.K., Neumann K. and Rothmund M. "Successful allotransplantation of microencapsulated parathyroids in rats." (1994) World Journal of Surgery, 18, 630-639.
75.Hesse C., Zielke A., Klock G., Barth P., Schlosser A., Zimmermann U., and Rothmund M., "First successful xenotransplantation of microencapsulated human parathyroid tissue in experimental hypoparathyroidism: long-term function without immunosuppression" (1997) J. Microencapsulation, 14, 617-626.
76.Jolley WB., Hinshaw DB., Call TW. and Alvord LS., "Xenogeneic pancreatic islet transplantation in proteolytic enzyme-bonded diffusion chambers in diabetic rats." (1977) Transplantation Proceedings. 9(1), 363-5.
77.Golden P. Greenwalt A. Martin K. Bellorin-Font E. Mazey R. Klahr S. and Slatopolsky E. "Lack of a direct effect of 1,25-dihydroxychole-calciferol on parathyroid hormone secretion by normal bovine parathyroid glands." (1980) Endocrinology. 107(2), 602-7.
78.Darquy S. and Sun AM. "Microencapsulation of parathyroid cells as a bioartificial parathyroid. In vitro studies." (1987) ASAIO Transactions. : 33, 356-358.
79.Nyberg SL., Remmel RP., Mann HJ., Peshwa MV., Hu WS. and Cerra FB., "Primary hepatocytes outperform Hep G2 cells as the source of biotransformation functions in a bioartificial liver." (1994) Annals of Surgery.：220(1), 59-67.
80.于仲元主編，高滬濱、田鬆、陶放副主編，（1993）血液淨化，淑馨出版社，金名圖書公司。
81.林杰樑編著，（1991）透析治療學，和記圖書出版社發行。
82.譚柱光編著，（1988）人工腎臟，金銘圖書有限公司。
83.李建賢著，彭芳谷校閱，（1993）副甲腺外科學，國立編譯館主編，日月文書局印行。
84.李豐編著，（1977）從甲狀腺腺腫談到人體賀爾蒙，89-101頁，中正書局印行。
85.中國生物科學編譯社編著，（1974）實驗動物解剖圖說，中國書局印行。