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姓名 黃美鳳(Mei-Feng Huang) 查詢紙本館藏 畢業系所 化學工程與材料工程學系 論文名稱 幾丁聚醣接枝半乳糖簇之材料性質及其肝靶向性研究
(Meterials characters and liver targer to study of chitosan graft galactose)相關論文 檔案 [Endnote RIS 格式]
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摘要(中) 本實驗研究，以幾丁聚醣和半乳糖醛酸以共價聯接形成一個接枝半乳糖基的幾丁聚醣薄膜和奈米粒子；另以L-lysine作為中間體，以L-lysine上的胺基和半乳糖醛酸上羧基之凝結回應合成結束簇狀(cluster)接枝半乳糖基之幾丁聚醣薄膜和奈米粒子；分別針對其物化性(拉力、抗菌等等)、Gal-1吸附能力、CLSM(Confocal scanning Microscope)觀察之肝臟細胞標靶測試分析。
薄膜實驗中顯示，當幾丁聚醣接枝半乳糖醛酸後，明顯的可以提升膜材對於E.coil 的抑菌能力，其因半乳糖基為清水基團可增加抗菌能力；接枝半乳糖基之幾丁聚醣薄膜較原始未接枝之幾丁聚醣薄膜明顯具有對Gal-1較佳之結合效能；肝細胞貼附型態模式也都因半乳糖基與肝細胞表面受體ASGP-R之間的半乳糖基特化辨識(galactose-specific recognition)主導了肝細胞在材料表面之貼附型態。
奈米粒子實驗其奈米粒子約300~500nm，且相對的zeta-potential，有接枝lysine之幾丁聚醣奈米粒子或簇狀接枝半乳糖基團之幾丁聚醣奈米粒子均明顯具有較高；而奈米粒子方面，其型態受了三聚磷酸之濃度及pH值之控制；最後以奈米粒子在固定時間點以CLSM(Confocal scanning Microscope)觀察之肝細胞標靶試驗，發現標的試驗之時間越長，螢光越明顯，且觀察到簇狀接枝半乳糖基團之幾丁聚醣奈米粒子都比幾丁聚醣及單點接枝半乳糖基之幾丁聚醣奈米粒子標的至肝細胞的數目明顯為多，作為奈米粒子標的至肝臟細胞之效率評估及佐證。摘要(英) In this study, the synthesis and characterization of a novel galactosyl chitosan are reported. The novel galactosyl chitosan was prepared by covalent coupling of lactobionic acid with chitosan(gal-chit) ; and galactosyl chitosan was prepared by introducing a L-lysine spacer to chitosan,followed by covalent coupling of lactobionic acid with the lysine spacer on chitosan(gal-lys-chit) ,and witch maked into membranes and nano-particles experiment.
Morphology and mechanical properties of membranes were studied by FTIR analysis, X-ray diffraction, swelling study, antibacterial test, bind galectin-1 and cell train. Nano-particles were studied by FTIR analysis, X-ray diffraction ,nano-particles sizes , zeta potential and target to liver cells.
In the film experiment, improve the ability to resist fungus of the gal-chit and gal-lys-chit membrane materials, because the lactobionic acid is base group of the hydrophile; and the ability to effectively bind galectin-1(Gal-1); galactose and surface receptor of liver cell can lead sticking to the growing type attitude on the material of liver cell.
In the nano-particles experiment, gal-chit,gal-lys,gal-lys-chit of the value are relatively high of the nano-paticles and zeta potential ,because three dimensional obstacle of the galactose of ring structure will influence effort of the electric charge between positive electricity amine base on the chitosan and negative electricity of TPP.The nano-particles form is affected by PH value and concentration of TPP.Nano-particles at some of set time observed to target cells of liver, the longer time is gal-lys-chit NPs has more apparent result than gal-chit NPs. Can be as efficiency assessing and evidence of enduring liver cell that the rice particle marks.關鍵字(中) ★ 半乳糖
★ 幾丁聚醣關鍵字(英) ★ chitosan
★ galactose論文目次 中文摘要…………………………………………………………………….I
英文摘要…………………………………………………………………...III
目錄………………………………………………………………………...VI
圖目錄………………………………..………………………………...…..XI
表目錄……………………………………………………………………XIV
第一章 绪論............................................................................................1
第二章 文獻回顧..................................................................................3
2.1幾丁聚醣簡介..................................................................................3
2.1.1幾丁聚糖..................................................................................3
2.1.2幾丁聚醣於生醫材料上之特色與應用.................................4
2.1.3幾丁聚醣奈米粒子在薬學上之應用.....................................5
2.2 Gal-1之簡介....................................................................................6
2.3奈米藥物載體………………………………………………………7
2.4奈米藥物載體之材料………………………………………………8
2.5 肝細胞...............................................................................................9
2.6去唾液酸胎醣蛋白接受器(ASGP-R)和半乳糖..................10
VI
第三章 薄膜實驗................................................................................14
3.1實驗目的.........................................................................................14
3.2實驗藥品.........................................................................................15
3.3實驗儀器設備...............................................................................16
3.4實驗方法.........................................................................................17
3.4.1膜材的製備..........................................................................17
3.4.1.1 戊二醛交聯幾丁聚醣薄膜的製備.............................17
3.4.1.2單點接枝半乳糖基之幾丁聚醣薄膜的製備.............18
3.4.1.3簇狀接枝半乳糖基之幾丁聚醣薄膜的製備.............19
3.4.2 紅外線光譜分析(FTIR)實驗...............................................21
3.4.3 X-ray光譜分析(XRD)實驗...................................................22
3.4.4膨潤度實驗............................................................................22
3.4.5抗菌實驗................................................................................23
3.4.6拉力測試實驗........................................................................24
3.4.7接枝率實驗............................................................................24
3.4.8 吸附GAL-1之效率..............................................................26
3.4.9 細胞實驗...............................................................................28
3.4.9.1電子顯微鏡觀察細胞的生長情形..............................28 VII
3.4.9.2 MTT assay...................................................................30
3.5結果與討論.....................................................................................32
3.5.1紅外線光譜分析(FTIR).......................................................32
3.5.2 X-ray光譜分析(XRD)分析...................................................33
3.5.3拉力測試分析........................................................................34
3.5.4膨潤性質分析........................................................................36
3.5.5抗菌分析................................................................................38
3.5.6接枝率分析............................................................................39
3.5.7 GAL-1之吸附分析...............................................................40
3.5.8 細胞.......................................................................................42
3.5.8.1倒立式光學顯微鏡觀察細胞的生長情形.................42
3.5.8.2MTT分析......................................................................47
第四章 奈米粒子實驗.....................................................................48
4.1實驗目的：.....................................................................................48
4.2實驗藥品.........................................................................................48
4.3實驗儀器設備................................................................................49
4.4實驗方法.........................................................................................50
4.4.1膜材的製備............................................................................50 VIII
4.4.1.1 幾丁聚醣奈米粒子之製備.........................................50
4.4.1.2 單點接枝半乳糖基之幾丁聚醣奈米粒子的製備....51
4.4.1.3 簇狀接枝半乳糖基之幾丁聚醣奈米粒子的製備....53
4.4.2 紅外線光譜分析(FTIR)實驗..............................................56
4.4.3 X-ray光譜分析(XRD)實驗....................................................56
4.4.4 動態散射式粒徑分析儀DLS（Dynamic Light Scattering）.........................................................................................................56
4.4.5 Zeta potential.........................................................................57
4.4.6示差掃瞄式熱分析儀DSC....................................................57
4.4.7 場發式電子顯微鏡..............................................................57
4.4.8 奈米粒子之肝細胞標靶實驗..............................................58
4.5結果與討論.....................................................................................59
4.5.1紅外線光譜分析(FTIR)分析................................................59
4.5.2 X-ray光譜分析(XRD)分析....................................................61
4.5.3粒徑分析................................................................................62
4.5.4 介面電位分析Zeta potential................................................65
4.5.5示差掃瞄熱分析儀DSC分析.................................................67
4.5.6場發式電子顯微鏡...............................................................68
4.5.7 奈米粒子之肝細胞標靶實驗..............................................71
第五章結論.............................................................................................74
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