臺灣博碩士論文加值系統

聚己內酯(Polycaprolactone, PCL)是普遍使用的生醫材料，但由於低親水性表面相對其他材料蛋白質與細胞貼附的效率較低，此外如同其他材料容易遭受細菌的汙染本研究以3,4,5-Trihydroxybenzaldehyd (THBA)與AMN的聚合物塗層改善PCL改善親水性，並搭配帶有醛基的聚乙烯醇和幾丁聚醣改良塗層抑菌效果。以XPS、SEM、水接觸角和表面電位做表面分析，以MTT實驗觀察細胞毒性和Live/Dead觀察材料抑菌效果。結果顯示AMN聚合物塗層改善PCL表面親水性，且無細胞毒性，但是PEG-Aldehyde無法顯著改善塗層表面抑菌性，然而藉由表面電位顯示幾丁聚醣吸附於表面，並且增強抑菌效果。本實驗結果確認胺基丙二氰聚合物搭配幾丁聚醣可應用於PCL表面改質親水性以及提升抑菌之可行性。

Polycaprolactone (PCL) is one of the biomaterials used in a variety of biomedical applications. The efficiency of protein or cell attachment were lower on PCL compared with other biomaterials; therefore, numerous PCL surface modification studies have been developed.In this study, we use 3,4,5-tirhydroxybenzaldehyde and Amino-malononitrile p-toluenesulfonate dissolved in alkalic water solution to improve hydrophilicity of PCL surface. Meanwhile, PEG-aldehyde or chitosan was added to AMN layer to improve antibacterial property. XPS, contact angle, SEM and MTT were used to determine the characters of modified surface of PCL and Live/Dead staining to measure the antibacterial activity. The result show that AMN coating successfully improve hydrophilicity of PCL surface; however, PEG-Aldehyde coating can’t significantly improve antibacterial property. On the contrary, chitosan plus AMN coating layer shows good antibacterial property.

目錄
指導教授推薦書
口試委員審定書
致謝 iii
摘要 iv
Abstract v
目錄 vi
圖目錄 ix
表目錄 xi
縮寫目錄 xii
第一章 緒論 - 1 -
第二章 文獻回顧 - 3 -
2-1.材料表面親水性對細胞貼附影響 - 3 -
2-2. Aminomalononitrile介紹 - 7 -
2-3.Aminomalononitirle應用於表面改質 - 9 -
2-4.材料表面細菌汙染 - 15 -
2-5聚乙二醇應用於抑菌貼附 - 17 -
2-6 幾丁聚醣應用於抑菌貼附 - 21 -
2-7 研究動機與目的 - 25 -
第三章 材料與方法 - 26 -
3-1.實驗藥品 - 26 -
3-2.使用儀器 - 28 -
3-3 實驗方法 - 29 -
3-3-1.製備PCL膜 - 29 -
3-3-2.製備AMN聚合物塗層 - 29 -
3-3-3.幾丁聚醣吸附層製備 - 30 -
3-3-4.樣品表面之特性分析 - 30 -
3-3-5. 細胞毒性實驗 - 30 -
3-3-6.金黃色葡萄球菌培養 - 32 -
3-3-7.抑菌測試實驗 - 32 -
3-3-8. 統計分析 - 33 -
第四章 結果與討論 - 34 -
4-1. SEM影像觀察 - 34 -
4-2. 表面元素分析 - 36 -
4-3.表面電位量測 - 38 -
4-4.細胞毒性測試 - 39 -
4-5.表面親水性測試 - 40 -
4-6.抑菌測試實驗 - 43 -
第五章 結論 - 46 -
參考文獻 - 47 -
 
圖目錄
圖2-1：CHO、MEF、BPAEC細胞於PE表面SEM影像。[32] - 4 -
圖2-2：三種塗層HUVECs貼附量與水接觸角關係圖。[33] - 5 -
圖2-4：以AMN合成氫化氰聚合物路徑圖。[35] - 7 -
圖2-5：AMN與(1)丙烯腈(2)乙醛酸反應與酸水解化學反應式。[37] - 9 -
圖2-6：表面XPS檢測N/C元素比值結果。[24] - 10 -
圖2-7 : AMN塗層於不同底材表面水接觸角結果。[24] - 11 -
圖2-8：AMN塗層細胞貼附實驗。[24] - 11 -
圖2-9：AMN塗層細胞毒性實驗。[24] - 12 -
圖2-10：(a)TFEA以及(b)TFAEH濃度與AMN聚合物F/C元素比關係圖。[24] - 12 -
圖2-11：於(a)第一天加入含有E.coli去離子水(b)隔一個禮拜加入純去離子水經過過濾器並檢測過濾後水中菌濃度結果圖。[38] - 14 -
圖2-12：材料表面細菌汙染流程示意圖。[44] - 16 -
圖2-13：聚乙二醇結構式。 - 17 -
圖2-14：(a)底材(b)聚多巴胺(c)PEG catechol塗層表面接觸角。[48] - 18 -
圖2-15：Amphora coffeaeformis貼附於(i)底材(ii)聚多巴胺(iii)PEG catechol塗層之(a)螢光影像與(b)量化數據結果。[48] - 18 -
圖2-16：PEG polymer brush製備於PDMS表面流程圖。[49] - 19 -
圖2-17：(a,b)E.coli與(c,d) S. aureus貼附於(a,c)PDMS與(b,d)PEG brush表面SEM影像以及(e)量化數據結果。[49] - 19 -
圖2-18：PEG分子量對水接觸角與細菌貼附量影響曲線。[50] - 20 -
圖2-19：幾丁質脫乙醯基化[51]。 - 21 -
圖2-20：S. aureus於不同表面鈦金屬表面存活度量化圖。[54] - 22 -
圖3-1：細胞代謝MTT產生紫色結晶示意圖。 - 31 -
圖4-1：(A)PCL (B) TAPCL (C)3P7T-APCL (D) 5P5T-APEG (E) PAPCL (F)1C-TAPCL (G) 3C-TAPCL 之SEM影像圖。(放大倍率：300倍，scare bar為100 µm) - 35 -
圖4-2：PEG-Aldehyde結構式 - 37 -
圖4-3：表面電位量測結果圖。 - 38 -
圖4-4：細胞毒性測試實驗結果。虛線為70 %基準線。 - 39 -
圖4-5：(A)PCL (B)TAPCL (C) 3P7T-APCL (D)5P5T-APCL (E) PAPCL (F) 1C-TAPCL (G) 3C-TAPCL Live & Dead螢光染色影像圖。（放大倍率：200x，Scale bar為100 µm） - 45 -
圖4-6：Live & Dead螢光染色量化直方圖。 - 45 -

 
表目錄
表2-1：各樣品表面水接觸角與XPS檢測結果。[34] - 6 -
圖2-3：NIH 3T3細胞於表面含有(A)5%血清 (B)2.1mg/ml BSA的不同種類樣品上細胞貼附情形。[34] - 6 -
表2-2：胺基丙二氰聚合物水解產物。[36] - 8 -
表2-3：AMN聚合物塗層表面元素分析與物理吸附力。[25] - 13 -
表2-3：E.Coli與S.aureus貼附實驗結果。[55] - 23 -
表2-4：金屬奈米粒子對三種菌種抑菌圈實驗結果。[56] - 24 -
表4-1：XPS表面元素atomic concentration檢測結果 - 37 -
表4-2：SEM-EDS表面元素分析結果 - 37 -
表4-3：樣品接觸角影像圖 - 42 -

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