多醣體改質poly(butyleneadipate-co-terephthalate)表面對其血液相容性、生物相容性之影響

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研究生：	謝瑞元 JUI YUAN HSIEH
論文名稱：	多醣體改質poly(butyleneadipate-co-terephthalate)表面對其血液相容性、生物相容性之影響 Effect of Surface modification with polysaccharides on the hemocompatibilityand Biocompatibility of poly(butyleneadipate-co-terephthalate)
指導教授：	楊銘乾 Ming-Chien Yang
口試委員:	李振綱 Cheng-Kang Lee 王大銘 none
學位類別：	碩士 Master
系所名稱：	工程學院 - 材料科學與工程系 Department of Materials Science and Engineering
論文出版年：	2006
畢業學年度：	94
語文別：	中文
論文頁數：	89
中文關鍵詞：	PBAT、幾丁聚醣、肝素、透明質酸、生物相容性、血液相容性
外文關鍵詞：	PBAT、chitosan、heparin、hyaluronic acid、biocom
相關次數：	點閱：289 下載：0
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本論文研究方向為以生物可分解共聚酯膜(poly(butyleneadipate-co-terephthalate)(PBAT)，利用臭氧活化方式，進行表面接枝丙烯酸（Acrylic Acid）於膜表面產生COOH基團，以共價鍵與幾丁聚醣(chitosan)結合，再以幾丁聚醣上的NH2與肝素（heparin）和透明質酸 (hyaluronic acid) 共價結合並且分別探討其表面改質後血液相容性、生物相容性、與L929纖維母細胞毒性，並且分析血液凝固性，以分析PBAT作為生醫材料之應用。

對於改質後PBAT薄膜的表面性質可利用XPS和染料分析出表面接枝的狀況。由AFM可觀察出薄膜經改質前後對表面粗糙度的影響，接枝肝素（heparin）和透明質酸(hyaluronic acid)可增加薄膜之親水性，經改質chitosan過後表面帶正電荷的聚酯與表面帶負電荷的蛋白質human serum albumin (HSA)或human plasma fibrinogen (HPF)其吸附量較接枝肝素（heparin）和透明質酸(hyaluronic acid)其聚酯表面為負電荷者高，測量APPT的結果發現接枝肝素的PBAT膜其凝血時間有大幅增長。另一方面經改質前後的PBAT薄膜以L929纖維母細胞觀察其生物毒性的結果可得PBAT薄膜未析出有毒物質影響L929纖維母細胞生長。

總結實驗結果改質後的PBAT有更好的生物相容性與血液相容性，其改質方式以共價鍵結有助於改質物固定在PBAT表面在發展成為生醫材料的領域上，有極大的潛力。

Acrylic acid was grafted to ozone treated Poly(butyleneadipate-co-
terephthalte) (PBAT) membranes. The resulting membranes were further grafted with chitosan (CS) via esterification. Afterward CS-grafted
membranes were immobilized with heparin (Hep) and hyaluronic acid (HA). The resulting PBAT membranes were characterized including the blood compatibility, the biocompatibility, and the cytotoxicity. These properties were employed to evaluate the applicability in biomedical areas.
Surface properties of immobilized PBAT membranes could be detected by XPS and dyeing. The roughness analysis between the grafted PBAT membranes and the ungrafted PBAT membranes could be measured by AFM. The effect of the positive charges of CS-grafted material on the absorption of negatively charged proteins of human serum albumin (HSA) and human plasma fibrinogen (HPF) was be studied. After grafting CS, the interaction between the membrane surface and the negatively charged proteins were electrostatic attraction . Thus the amount of adsorption /attachment increased with the surface positive charges on the membranes. One the other hand, the PBAT membranes immobilized with heparin and hyaluronic acid could decreased the amount of adsorption/attachment proteins. The heparin-grafted PBAT membrane showed longer APTT. In addition, the in vitro cytotoxicity test results from culturing fibroblasts (L929) show that these PBAT membranes were not cytotoxic.

目錄
頁數
中文摘要I
英文摘要II
誌謝III
目錄IV
圖表索引VII
第一章緒論1
1-1研究背景1
1-2研究目的2
第二章文獻與理論基礎3
2-1實驗材料簡介3
2-1-1生物分解性聚酯高分子材料3
2.1.2幾丁聚醣(Chitosan)之生物相容性及抗菌性5
2.1.3透明質酸(hyaluronic acid)之生物相容性與保水性7
2-1-4肝素（Heparin）9
2-2高分子材料表面改質(Surface modification)13
2-2-1DPPH 過氧化物分析17
2-3薄膜表面分析18
2-3-1接觸角19
2-3-2原子力顯微鏡之原理(AFM)21
2-3-3血液的成分23
2-4血液學凝固原理26
2-5
2-6凝血作用程序
凝血時間                                                                                                 28
30
第三章實驗材料與方法31
3-1實驗材料31
3-2實驗儀器32
3.3實驗流程33
3.4-1PBAT薄膜製備34
3-4-2臭氧處理35
3-4-3PBAT接枝丙烯酸(PBAT-g-PAA)36
3-4-4Chitosan(CS)固定化於改質的PBAT薄膜37
3-4-5透明質酸 (hyaluronic acid HA) 固定化於PBAT薄膜38
3-4-6肝素(Heparin ,Hep)固定化於PBAT薄膜39
3-5-1過氧化物測定(Peroxide determination)41
3-5-2表面接枝密度測定(Sufface Graft Density)42
3-5-3薄膜表面親水性43
3-5-4機械強度測試(Mechanical Test)43
3-5-5X射線光電子能譜儀43
3-5-6活性部分凝血時間 (APTT)44
3-5-7
3-5-8
3-5-9
3-5-10
3-5-11HAS和HSA蛋白質吸附
血小板貼附評估(Evaluation of platelet adhesion)
原子力顯微鏡(AFM)
細胞毒性
細胞增生45
45
46
47
50
第四章結果與討論47
4-1薄膜臭氧處理(Ozone treatment)52
4-2接枝密度的測量57
4-3射線光電子能譜儀-XPS (X-ray Photoelectron Spectroscopy－X)表面元素分析59
4-4 薄膜改質後之親水性63
4-5HAS 和HPF吸附65
4-6APTT  (Activated partial thromboplastin time)67
4-7血小板活化 ( Activation of platelets)68
4-8原子力顯微鏡—AFM70
4-9細胞毒性77
4-10細胞增生79
第五章結論(Conclusion)81
第六章參考文獻(Reference)82
作者簡介89














圖表索引
Figure 2-1Ecoflex®化學結構式3
Figure 2-2Chitin 即Chitosan化學結構式4
Figure 2-3hyaluronic acid化學結構式8
Figure 2-4肝素（Heparin）化學結構式9
Figure 2-5DPPH化學結構式17
Figure 2-6接觸角量測示意圖19
Figure 2-7血液凝結途徑29
Figure 3-1臭氧反應裝置35
Figure 3-2PBAT接枝丙烯酸(PBAT-g-PAA)40
Figure 3-3Chitosan(CS)固定化於改質的PBAT薄膜40
Figure 3-4透明質酸固(hyaluronic acid HA)定化於PBAT薄膜40
Figure 3-5肝素(Heparin,Hep)固定化於PBAT薄膜40
Figure 3-6活性部分凝血時間 (APTT) 測試流程44
Figure 4-1The effect of the peroxide density on the ozone treating time54
Figure 4-2The effect of the surface density of carboxylic group on the 55
ozone treating time.
Figure 4-3The dependence of the breaking strength of PBAT on the ozone56
treating time
Figure 4-4Compare the surface density of carboxylic group and amino group 58
on the ozone treating time.
Figure 4-5The surface density of functional groups grafted.58
Figure 4-6The XPS spectra wide scan of the PBAT membrane60
Figure 4-7The XPS spectra wide scan of the PBAT-CS-Hep membrane60
Figure 4-8The XPS spectra narrow scan of the PBAT-CS-Hep membrane61
Figure 4-9The XPS spectra narrow scan of the PBAT-CS-Hep membrane61
Figure 4-10The water contact angles of PBAT、PBAT-CS、PBAT-CS- Hep、64
PBAT-CS-HA membranes
Figure 4-11Amount of HPF and HAS absorbed onto  PBAT，PBAT-CS，66
PBAT-CS-HA，and PBAT-CS-Hep membrane surfaces
Figure 4-12Comparison of platelet adhesion and activation on untreated and  69
surfaced modified PBAT membranes
Figure 4-13(a) The Roughness analysis of PBAT membrane.72
Figure 4-13(b) AFM image of PBAT membrane.72
Figure 4-14(a)The Roughness analysis of PBAT-AA membrane.73
Figure 4-14(b)AFM image of PBAT-AA membrane.73
Figure 4-15(a)The Roughness analysis of PBAT-CS membrane.74
Figure 4-15(b) AFM image of PBAT-CS membrane.74
Figure 4-16(a) The Roughness analysis of PBAT- CS-HA membrane.75
Figure 4-16 (b) AFM image of PBAT- CS-HA membrane.75
Figure 4-17(a)The Roughness analysis of PBAT- CS-Hep membrane76
Figure 4-17 (b)AFM image of PBAT- CS-Hep membrane76
Figure 4-18The in vitro cytotoxicity test results of fibroblasts on (A)blank78
(B)DMSO，(C)PBAT，(D)PBAT-CS，(E)PBAT- CS-Hep，(F)PBAT- CS-HA
Figure 4-19 The in vitro cytotoxicity test results of PBAT、PBAT-CS、PBAT- CS-Hep  79
and PBAT- CS-HA  membranes.The initial concentration of fibroblasts
was 80000cell/ml.Fibroblasts  were incubated at 37℃for 3 days
Figure 4-20Cell growth ratios of PBAT, PBAT-CS, PBAT- CS-Hep and PBAT- CS-HA 80
membranes.The initial concentration of fibroblasts was 50000cell/ml. Fibroblasts were incubated at 37℃for 7days
Table 2-1各種表面處理法之比較                                         15
Table 4-1The atomic rate of  PBAT and PBAT- CS-Hep membranes.              62
Table 4-2PBAT, PBAT-CS ,PBAT-CS-Hep ,PBAT-CS-HA membranes of APTT         67
(activated partial thromboplastin time)
Table 4-3The mean roughness of PBAT membranes were measured by AFM.    70

                                

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