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研究生: 梁祥發
Xiang-Fa Liang
論文名稱: 以Lactobacillus reuteri菌發酵glycerol生成reuterin做為生物組織材料天然滅菌劑的探討
指導教授: 高振宏
C. Robert Kao
口試委員:
學位類別: 碩士
Master
系所名稱: 工學院 - 化學工程與材料工程學系
Department of Chemical & Materials Engineering
畢業學年度: 89
語文別: 中文
論文頁數: 65
中文關鍵詞: 滅菌劑reuterinLactobacillus reuteri生物組織材料
外文關鍵詞: reuterin, Lactobacillus reuteri, biological tiss
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  • 生物組織材料在臨床上,可以用來暫時性或永久性的取代病人缺損的組織或器官。這些生物組織材料在經過適當的交聯劑交聯處理後,可以減低這些異種生物組織材料來源的抗原性,並增加生物組織材料在植入人體後抵抗酵素分解的能力。然而這些經過交聯劑化學修飾的生物組織材料,在植入人體之前仍需經過適當的滅菌處理,以大量減少或完全去除材料上所帶有的微生物,避免病人遭受致病微生物的感染。然而目前臨床上對於生物組織材料,尚未有一個理想、適合的滅菌方法。
    本實驗室目前正著手研究一種新的天然滅菌劑-Reuterin。Reuterin是由人或動物的腸胃道所分離出的乳酸菌Lactobacillus reuteri在厭氧的狀態下發酵glycerol後所產生的滅菌物質。其抑菌作用機制可能與RNA氧化還原


    摘要Ⅰ 目錄Ⅲ 圖索引Ⅵ 第一章 緒論1 1.1 生物組織材料1 1.2 滅菌方法1 1.3天然滅菌劑reuterin2 1.3.1 Lactobacillus reuteri3 1.3.2 Reuterin3 1.4 研究動機與目的4 第二章 Lactobacillus reuteri發酵glycerol生成reuterin6 2.1 研究目的6 2.2 實驗材料6 2.2.1 Lactobacillus strain6 2.2.2 Reuterin的製備6 2.3 實驗方法8 2.3.1 Reuterin分離純化8 2.3.2 FT-IR、GC-MS及NMR分析9 2.3.3 L. reuteri選擇及鑑定10 2.3.4 L. reuteri發酵glycerol最適化條件的探討10 2.3..4.1 生長曲線10 2.3.4.2 發酵glycerol最適化條件探討12 2.4 實驗結果與討論13 2.4.1 Reuterin分離純化13 2.4.2 FT-IR、GC-MS及NMR分析14 2.4.3 乳酸菌選擇及鑑定18 2.4.4 L. reuteri發酵glycerol最適化條件的探討19 2.4.4.1 生長曲線19 2.4.4.2 發酵glycerol最適化條件探討21 2.5 結論25 第三章 Reuterin滅菌性質與細胞毒性測試26 3.1 研究目的26 3.2 實驗材料26 3.2.1 Reuterin製備26 3.2.2 Genipin交聯組織27 3.2.3 實驗菌株與細胞28 3.3 實驗方法29 3.3.1 Reuterin滅菌性質29 3.3.2 細胞毒性測試31 3.3.2.1 SEM觀察細胞生成情形31 3.3.2.2組織病理切片32 3.3.2.3 MTT assay34 3.4 實驗結果與討論35 3.4.1 Reuterin的滅菌性質35 3.4.2 細胞毒性測試39 3.4.2.1 SEM觀察細胞生成情形39 3.4.2.2組織病理切片40 3.4.2.3 MTT assay44 3.5 結論45 第四章Reuterin穩定性探討46 4.1 研究目的46 4.2材料與方法46 4.2.1 Reuterin製備46 4.2.2 實驗方法46 4.2.2.1 Reuterin溶液穩定性測試46 4.2.2.2 Reuterin降解後物質的滅菌性質以及細胞相容性47 4.3 實驗結果與討論51 4.3.1 Reuterin溶液穩定性測試51 4.3.2 Reuterin降解後的滅菌性質與細胞毒性測試54 4.4 結論58 參考文獻59 附件63

    1.M. E. Nimni, D. Cheung, B. Strates, M. Kodama, and K. Sheikh, “Bioprosthesis derived from cross-linked and chemically modified collagenous tissues,” in Collagen, Vol.Ⅲ, 1-38, CRC Press, Boca Raton, FL, 1998.
    2.J. P. Iguidbashian, D. M. Follette, and J. P. Contino, “Pericardial patch repair of left ventricular aneurysm,” Ann. Thorac. Surg., 55, 1022-1024, 1993.
    3.S. Gabbay, U. Boltolotti, and S. Factor, “Calcification of implanted xenograft pericardium: Influence of site and function,” J Thorac. Cardiovasc. Surg., 87, 782-787, 1984.
    4.Okoshi, Y. Noishiki, and Y. Tomizawa, “Development of an antithrombogenic cardiac wall substitute which can be reconstructed by infiltration of host cells,” Trans. Am. Soc. Artif. Intern. Organs, 34, 532-537, 1988.
    5.H. Kurosawa, Y. Imai, and M. Nakazawa, “Standardized patch infundibuloplasty for tetralogy of Fallot,” J. Thorac. Cardiovasc. Surg., 92, 396-401, 1986.
    6.D. J. Dempsey and R. P. Thirucote, “Sterilization of medical devices: a review,” J. Biomat. Appl., 3, 1989.
    7.E. A. Wooddroff, “Use of glutaraldehyde and formaldehyde to process tissue heart valves,” J. Bioeng., 2, 1-10, 1978.
    8.R. Trujillo and T. J. David, “Sporostatic and sporocidal properties of aqueous formaldehyde,” Appl. Microbiol., 23, 618-622, 1972.
    9.A. Carpentier, A. Deloche, J. Relland, N. Fabiani, J. Forman, J. P. Camilleri, R. Soyer, and R. Dubost, “Six-year follow-up of glutaraldehyde-preserved heterografts,” J. Thorac. Cardiovas. Surg., 68, 771, 1974.
    10.P. M. Borick, F. H. Dondershine, and V. L. Chandler, “Alkinized glutaraldehyde: A new antimicrobial agent,” J. Pharm. Sci., 52, 1273, 1964.
    11.J. M. Lohre, J. L. Baclig, S. Sagartz, K. T. Guida, and R. Tu, “Evaluation of two epoxy ether compounds for biocompatible potential,” Artif. Organs., 16, 630-633, 1992.
    12.J. M. Lohre, L. Baclig, E. Wickham, S. Guida, J. Farley, K. Thyagarajan, R. Tu, and R.C. Quijano, “Evaluation of epoxy ether fixed bovine arterial grafts for mutagenic potential,” ASAIO J., 39, 106-113, 1993.
    13.C. Nishi, N. Nakajima, and Y. Ikada, “In vitro evaluation of cytotoxicity of diepoxy compounds used for biomaterial modification,” J. Biomed. Mater. Res., 29, 829-834, 1995.
    14.D. P. Speer, M. Chvapil, C. D. Eskelson, and J. Ulreich, “Biological effects of residual glutaraldehyde in glutaraldehyde-tanned collagen biomaterials,” J. Biomed. Mater. Res., 14, 753-764, 1980.
    15.W. J. Dorogosz and S. E. Lindgren: U.S. Patent 5413960, 1995.
    16.S. E. Lindgren and W. J. Dorogosz, “Antagonistic activities of lactic acid bacteria in food and feed fermentation.” FEMS Microbiol. Rev., 87, 149-164, 1990.
    17.FDA, “Pasteurized milk ordinance. Publication No. 229, Public Health Service. Food and Drug Administration. U. S. Dept Health and Human Services.” 1989.
    18.M. L. Speck, W. J. Dobrogosz and I. A. Casas, “Lactobacillus reuteri in food supplementation.” Food Technol., 47, 90-94, 1993.
    19.T. L. Talarico, I. A. Casas, T. C. Chung and W. J. Dobrogosz, “Production and Isolation of Reuterin, a Growth Inhibitor Produced by Lactobacillus reuteri,” Antimicrob. agents chemother., 1854-1858, Dec 1988.
    20.T. L. Talarico and W. J. Dobrogosz, ”Chemical Characterization of Antimicrobial Substance Produced by Lactobacillus reuteri,” Antimicrob. agents chemother., 674-679, May 1989.
    21.M. G. El-Ziney, T. van den Tempel, J. M. Debevere and M. Jakobsen, “Application of reuterin produced by Lactobacillus reuteri 12002 for meat decontamination and preservation.” J. food prot., 62, 3, 257-261, 1999.
    22.陳勁初, “以乳酸菌保存食品之機制.” 食品工業, 23, 9, 17-21, 1999.
    23.M. G. El-Ziney and J. M. Debevere, “The effect of reuterin on Listeria monocytogenes and Escherichia coli O157:H7 in milk and cottage cheese.” J. food prot., 61, 10, 1275-1280, 1998.
    24.H.-W. Sung, R.-N. Huang, L.L.-H. Huang and C. C. Tsai, “In vitro evaluation of cytotoxicity of a naturally occurring cross-linking reagent for biological tissue fixation.” J. Biomater. Sci. Polymer Edn., 10, 1, 63-78, 1999.
    25.H.-W. Sung, Y. Chang, C.-T. Chiu, C.-N. Chen and H. C. Liang, “Crosslinking characteristics and mechanical properties of a bovine pericardium fixed with a naturally occurring crosslinking agent.” J. Biomed. Mater. Res., 47, 116-126, 1999.
    26.H.-W. Sung, Y. Chang, C.-T. Chiu, C.-N. Chen and H.-C. Liang, “Mechanical properties of a porcine aortic valve fixed with a naturally occurring crosslinking agent.” Biomaterials., 20, 1759-1772, 1999.
    27.H.-W. Sung, C.-C. Tsai and R.-N. Huang, “Extraction of a naturally occurring crosslinking agent (genipin) from Gardenia fruits and its applications in biological tissue fixation.” Biomaterials and Drug Delivery toward New Mellenium, 623-632, 2000.
    28.M. L. Shuler and F. Kargi, “Bioprocess engineering basic concepts.” Prentice Hall P T R, Englewood Cliffs, New Jersey.
    29.陳俊男,博士論文,中央大學化工所,2001.
    30.蔡文城,微生物學,第三版,藝軒圖書出版社,1996.
    31.H. B. Chan-Myers, S. H. Guida, C. G. Roberts, K. Thyagarajan, R. Tu and R. C. Quijano, “Sterilization of a small caliber vascular graft with a polyexpoxy compound — Denacol.” Baxter Technical Jouunal.
    32.A. Balows, W. J. Hausler and J. P. Truant, “Manual of clinical microbiology.” 3rd edition, American Society for Microbiology, Washington, D.C. 1980.
    33.G. Herman and J. R. Richey, “Fundamentals of Organic Chemistry.” New Jersey, Prentice-Hall, ch. 9, 247-271, 1983.
    34.Manasses. K. Yunmbam and J. F. Roberts, “In vivo evaluation of reuterin and its combinations with suramin, melarsoprol, DL-α-difluoromethylornithine and bleomycin in mice infected with Trypanosoma brucei brucei.” Comp. Biochem. Physiol., 105c, 3, 521-524, 1993.

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