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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊台鴻(Tai-Horng Young) | |
| dc.contributor.author | "Lin, Yu-Hsiang" | en |
| dc.contributor.author | 林鈺翔 | zh_TW |
| dc.date.accessioned | 2021-06-13T01:07:30Z | - |
| dc.date.available | 2007-07-27 | |
| dc.date.copyright | 2007-07-27 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-23 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/29453 | - |
| dc.description.abstract | 本實驗是以乾式法將高分子乙烯乙烯醇(EVAL)聚合物製成薄膜之後，用化學改質的方式將表面氫氧基(-OH)接枝甲基丙烯醯氯(MAC)以間位氯化過氧苯酸(mCPBA)將烯烴雙鍵上進行環氧化反應化後接枝離胺酸(L-lysine)單體和各式不同碳鏈的雙胺。並且將接枝離胺酸(L-lysine)後的薄膜用碳化二亞胺(EDC)修飾離胺酸上的羧酸基接上丁二胺，並利用FT-IR以及接觸角變化分析薄膜改質前後差異和Orange II salt定量膜材表面胺基(-NH2)之接枝密度。
同時利用體外細胞培養的方法，來探討材料與細胞之間的生物適合性，將改質前以及改質後的薄膜培養小鼠初代顆粒型神經細胞，以MTT測試來比較細胞的活性並且用光學顯微鏡和掃描式電子顯微鏡來觀察細胞貼附在接枝不同碳鏈的二胺、離胺酸以及經丁二胺修飾之離胺酸並和塗佈聚離氨酸(poly-D-lysine)的乙烯乙烯醇薄膜做比較。 小鼠顆粒型神經細胞培養於接枝在經丁二胺修飾過後的lysine其細胞活性表現優於EVAL塗佈聚離胺酸薄膜更勝於未改質薄膜，效果比接枝雙胺類更能提升神經細胞粒線體活性。 | zh_TW |
| dc.description.abstract | We prepare poly(ethylene-co-vinyl alcohol)(EVAL) membranes by the dry processing ,and then grafted methacryloyl chloride (MAC)on the hydroxyl group of these membrane surface by the way of chemical modification.Furthermore, we grafted different diamines, lysine and modified lysine with 1,4-diaminobutane by EDC/NHS method on them.
We used FT-IR and contact angle to assay the material properties after modification then quantification the surface grafted density by orange II salt. We cultured the cerebellar granule neurons of the Wistar rat on the modified membranes and compared with the relative activity by MTT assays. In addition, the cell morphology was observed by optical microscope and scanning electron microscope (SEM). The modification of the EVAL with lysine modified 1, 4-diamino -butane, in general, have better relative activity than coating PDL. We observe this method could effectively induce the growth of neuron cells. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T01:07:30Z (GMT). No. of bitstreams: 1 ntu-96-R94548035-1.pdf: 4397896 bytes, checksum: e1525a30ad91c83925913df4eb856f65 (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 誌謝 I
中文摘要 II Abstract III 目錄 IV 圖表目錄 VII 第1章 緒論 1 第2章 文獻回顧 3 2-1高分子薄膜 3 2-1-1高分子薄膜定義 3 2-1-2高分子薄膜結構 3 2-1-3薄膜的製備 3 2-2神經細胞培養 4 2-2-1神經系統簡介 5 2-2-2培養神經細胞用途 6 2-2-3影響神經細胞生長因素 7 2-3薄膜改質 8 2-3-1改質簡介 8 2-3-2材料表面改質與細胞生長的關係 9 第3章 實驗材料與方法 11 3-1實驗架構 11 3-2試藥原料 11 3-3儀器 13 3-4試藥配置 18 3-5薄膜製備 21 3-5-1EVAL薄膜製備 21 3-5-2薄膜表面改質 21 3-5-2-1EVAL高分子薄膜接枝不同碳數雙胺和離胺酸單體 21 3-5-2-2 丁二胺修飾接枝離胺酸之EVAL高分子薄膜 22 3-6薄膜表面親疏水性質 26 3-7材料表面化學鑑定 27 3-7-1衰減全反射式傅立葉紅外光譜儀(ATR-FTIR) 27 3-8薄膜表面接枝雙胺量分析 27 3-9神經細胞培養 28 3-9-1薄膜前處理 28 3-9-2塗佈聚離胺酸 28 3-9-3初代細胞培養(primary cell culture)步驟 29 3-9-4 MTT測試評估神經細胞生長情形 30 3-10 電子顯微鏡 30 3-10-1細胞固定 31 3-10-2零界點乾燥 32 3-10-3樣本鍍金 33 第4章 結果與討論 34 4-1薄膜表面改質材料分析 34 4-1-1衰減全反射傅立葉紅外光譜儀(ATR-FTIR)分析 34 4-1-2薄膜表面接枝雙胺量分析 35 4-1-3薄膜表面親疏水性 36 4-2小鼠顆粒型神經細胞培養 37 4-2-1 EVAL接枝各式雙胺、離胺酸、修飾羧酸基之離胺酸細胞活性MTT測試 37 4-2-2神經細胞在不同材料的光學顯微鏡和電子顯微鏡(SEM)照片， 39 4-2-3神經細胞在不同材料的光學顯微鏡照片，細胞密度 2.5 x 105 cells/cm2。 40 第5章 結論 42 第6章 參考文獻 43 附錄 51 圖3-2 EVAL薄膜化學改質流程圖 23 圖3-3 EVAL接枝雙胺類和離胺酸單體之步驟 24 圖3-4 EVAL接枝丁二胺修飾離胺酸之步驟 25 圖3-5 接觸角示意圖 26 圖1-1 EVAL薄膜改質接枝不同胺基示意圖 54 圖3-1 實驗流程圖 54 圖3-6 初代小腦顆粒型神經細胞動物實驗步驟 56 圖4-1. EVAL接枝甲基丙烯醯氯(MAC)對時間(t)吸收強度 56 圖4-2. EVAL改質前後之ATR-FTIR圖 57 圖4-3. EVAL接枝不同胺類之ATR-FTIR圖 57 圖4-4各種改質EVAL薄膜培養小腦顆粒型神經細胞MTT測試結果 60 圖 4-5 顆粒型神經細胞初代培養第一天之光學顯微鏡照片；200x。 61 (a).EVAL，(b). PDL，(c). E-Lysine，(d). E-Ly-1, 4-diaminobutane 61 (e).E-1, 2-diaminoethylene，(f). E-1, 3-diaminopropane， 62 (g).E-1, 4-diaminobutane， (h). E-1, 5-diaminopetane， 62 圖 4-7顆粒型神經細胞初代培養第一天之光學顯微鏡照片；200x。 62 (i).E-1, 5-diaminohexane 62 圖 4-8 顆粒型神經細胞初代培養第三天之光學顯微鏡照片；200x。 63 (a).EVAL，(b). PDL，(c). E-Lysine，(d). E-Ly-1, 4-diaminobutane 63 圖 4-9 顆粒型神經細胞初代培養第三天之光學顯微鏡照片；200x。 64 (e).E-1, 2-diaminoethylene，(f). E-1, 3-diaminopropane， 64 (g).E-1, 4-diaminobutane， (h). E-1, 5-diaminopetane 64 圖 4-10 顆粒型神經細胞初代培養第三天之光學顯微鏡照片；200x。 64 (i).E-1, 5-diaminohexane 64 圖 4-11 顆粒型神經細胞初代培養第五天之光學顯微鏡照片；200x。 65 (a).EVAL，(b). PDL，(c). E-Lysine，(d). E-Ly-1, 4-diaminobutane 65 圖 4-12 顆粒型神經細胞初代培養第五天之光學顯微鏡照片；200x。 66 (e).E-1, 2-diaminoethylene，(f). E-1, 3-diaminopropane， 66 (g).E-1, 4-diaminobutane， (h). E-1, 5-diaminopetane， 66 圖 4-13 顆粒型神經細胞初代培養第五天之光學顯微鏡照片；200x。 66 (i).E-1, 5-diaminohexane 66 圖 4-14 顆粒型神經細胞初代培養第一天之SEM照片；500x。 68 (a). EVAL，(b). PDL，(c). E-Lysine，(d). E-Ly-1,4-diaminobutane， 68 (e).E-1, 2-diaminoethylene，(f). E-1, 3-diaminopropane， 68 (g). E-1, 4-diaminobutane， (h). E-1, 5-diaminopetane， 68 (i). E-1, 5-diaminohexane 68 圖 4-15 顆粒型神經細胞初代培養第三天之SEM照片；500x。 70 (a).EVAL，b. PDL，(c). E-Lysine，(d). E-Ly-1, 4-diaminobutane， 70 (e).E-1, 2-diaminoethylene，(f). E-1, 3-diaminopropane， 70 (g).E-1, 4-diaminobutane， (h). E-1, 5-diaminopetane， 70 (i).E-1, 5-diaminohexane 70 圖 4-16 顆粒型神經細胞初代培養第三天之SEM照片；500x。 72 (a).EVAL，(b). PDL，(c). E-Lysine，(d). E-Ly-1, 4-diaminobutane， 72 (e).E-1, 2-diaminoethylene，(f). E-1, 3-diaminopropane， 72 (g).E-1, 4-diaminobutane， (h). E-1, 5-diaminopetane， 72 (i).E-1, 5-diaminohexane 72 圖4-17小腦顆粒型神經細胞培養在EVAL。 73 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 73 (d) 7 DIV _ 200x 73 圖4-18小腦顆粒型神經細胞培養在EVAL-PDL 。 74 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 74 (d) 7 DIV _ 200x 74 圖4-19小腦顆粒型神經細胞培養在EVAL-Ly 。 75 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 75 (d) 7 DIV _ 200x 75 圖4-20小腦顆粒型神經細胞培養在EVAL-diaminobutane 。 76 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 76 (d) 7 DIV _ 200x 76 圖4-21小腦顆粒型神經細胞培養在EVAL-diaminoethylene 。 77 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 77 (d) 7 DIV _ 200x 77 圖4-22小腦顆粒型神經細胞培養在EVAL-diaminopropane 。 78 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 78 (d) 7 DIV _ 200x 78 圖4-23小腦顆粒型神經細胞培養在EVAL-diaminobutane 。 79 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200x、(c) 7 DIV _ 100x、 79 (d) 7 DIV _ 200x 79 圖4-24小腦顆粒型神經細胞培養在EVAL-diaminopantane 。 80 (a)1 DIV _ 200x 、(b) 7 DIV _ 200x 80 圖4-25小腦顆粒型神經細胞培養在EVAL-diaminohexane 。 80 (a.) 1 DIV _ 100X、(b) 1 DIV _ 200X、(c) 7 DIV _ 100X、 80 (d) 7 DIV _ 200ｘ 80 圖4-26小腦顆粒型神經細胞培養在各種改質材料第25天 81 (a)EVAL、(b)E-PDL、(c)E-Ly、(d)E-Ly-diamine(4) 、(e)E-diamine(2) 、 81 (f)E-diamine(3) 、(g)E-diamine(4) 、(h)E-diamine(5) 、(i)E-diamine(6) 81 表4-1 ATR-FTIR光譜吸收峰 34 表4-2不同濃度之OrangeⅡ於479nm的UV光之校正曲線 58 表4-3 EVAL薄膜接枝雙胺量之分析 58 表4-4接枝各式雙胺之EVAL薄膜表面接觸角 59 | |
| dc.language.iso | zh-TW | |
| dc.subject | 聚離胺酸 | zh_TW |
| dc.subject | 聚乙烯乙烯醇 | zh_TW |
| dc.subject | 小腦顆粒神經細胞 | zh_TW |
| dc.subject | 甲基丙烯醯氯 | zh_TW |
| dc.subject | 間位氯 | zh_TW |
| dc.subject | 化過氧苯酸 | zh_TW |
| dc.subject | 碳化二亞 | zh_TW |
| dc.subject | cerebellar granule cells | en |
| dc.subject | lysine | en |
| dc.subject | EVAL | en |
| dc.subject | 4-diaminobutane | en |
| dc.subject | PDL | en |
| dc.subject | EDC | en |
| dc.subject | methacryloyl Chloride | en |
| dc.title | 利用聚乙烯乙烯醇薄膜接枝雙胺探討其對神經元細胞之影響 | zh_TW |
| dc.title | The Study of the Behavior of Neurons Cultured on EVAL
Membranes Modified With Diamines | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 宋信文,謝松蒼 | |
| dc.subject.keyword | 聚乙烯乙烯醇,小腦顆粒神經細胞,甲基丙烯醯氯,間位氯,化過氧苯酸,碳化二亞,聚離胺酸, | zh_TW |
| dc.subject.keyword | cerebellar granule cells,methacryloyl Chloride,EDC,PDL,1,4-diaminobutane,EVAL,lysine, | en |
| dc.relation.page | 81 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-07-23 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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