電紡絲製備幾丁聚醣/褐藻酸鈉之奈米複合纖維結構對細胞貼附與增生之研究

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研究生：	余興寧 Hsing-ning Yu
論文名稱：	電紡絲製備幾丁聚醣/褐藻酸鈉之奈米複合纖維結構對細胞貼附與增生之研究 Electrospinning of Chitosan/Alginate Composite Nanofibrous Structures for Cell Adhesion and Proliferation
指導教授：	胡威文 Wei-wen Hu
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：	100
語文別：	中文
論文頁數：	98
中文關鍵詞：	細胞、褐藻酸鈉、幾丁聚醣、電紡絲
外文關鍵詞：	electrospinning, chitosan, alginate, cell
相關次數：	點閱：14 下載：0
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為了精確調控生物材料表面性質以利於組織工程的應用，我們研
發了利用電紡絲技術製備幾丁聚醣/褐藻酸鈉之複合纖維。首先，先
進行純幾丁聚醣與純褐藻酸鈉的電紡絲研究。聚氧化乙烯(PEO)分別
摻入於幾丁聚醣與褐藻酸鈉溶液中，並藉由掃描式電子顯微鏡(SEM)
研究高分子組成對於纖維型態的影響。利用高分子溶液的黏度分析證
實了高濃度的高分子造成的高黏度，影響纖維絲結構的型態。之後我
們研究進料流率與纖維沉積的關係，藉由控制幾丁聚醣/聚氧化乙烯
和褐藻酸鈉/聚氧化乙烯溶液的進料流率，可得到不同沉積比的奈米
纖維。最後，所合成的複合奈米纖維被應用於細胞培養，因細胞在幾
丁聚醣與褐藻酸鈉纖維絲的型態不同，我們可以透過奈米纖維的比例
調控細胞的型態。在生物相容性上，發現幾丁聚醣與褐藻酸鈉的奈米
纖維可以改善膜的細胞活性，此外複合纖維甚至比幾丁聚醣與褐藻酸
鈉纖維絲有更好的生物相容性。這些結果表示，奈米纖維的比例可以
透過同時紡絲技術加以控制，我們能夠操控複合材料的表面特性，這
應有利於在生物材料的應用。

To custom-tailor biomaterial surfaces for tissue engineering
application, we developed chitosan/alginate composite surfaces using
electrospinning technique. Firstly, chitosan and alginte were solely
electrospun, respectively. Poly (ethylene oxide) (PEO) was added to both
chitosan and alginte solutions, and the effect of polymer composition on
fibrous morphology was studied through scanning electrical microscopy
(SEM). In addition, viscosities of polymer solution were also analyzed,
demonstrating that higher concentrations of polymer caused higher
viscosities, which directly determined the morphology of nanofibrous
structure. Then, the relationship between flow rates and fiber deposition
was also studied. By controlling the flow rates of chitosan/PEO and
alginate/PEO solution separately, these nanofibers were able to be spun
simultaneously with different deposition ratios. Finally, the composite
nanofibrous surfaces were applied for cell culture. Because cell
morphologies were different in chitosan and alginate nanofibrous surfaces,
we may regulate cell morphologies by controlling the ratios of nanofibers,
About biocompatibility, it was found that nanofibers can improve the cell
viability over chitosan and alginate films. In addition, composite
nanofibers even demonstrated greater biocompatibility than that of
chitosan and alginate fibers. These results suggested that the ratio of
nanofiber can be finely controlled through simultaneous electrospinning
technique, and we may manipulate composite surface properties, which
should be beneficial to biomaterial application.

III
目錄
摘要................................................................................................................................ I
Abstract......................................................................................................................... II
目錄..............................................................................................................................III
圖目錄...........................................................................................................................V
第一章  緒論............................................................................................................1
1-1  背景............................................................................................................1
1-2  實驗目的....................................................................................................3
第二章  文獻回顧....................................................................................................5
2-1  組織工程....................................................................................................5
2-2  電紡絲於組織工程之應用........................................................................7
2-3  電紡絲簡介..............................................................................................11
2-3-1  電紡絲技術之發展......................................................................11
2-3-2  電紡絲之優點..............................................................................12
2-3-3  電紡絲原理..................................................................................12
2-3-4  影響電紡絲之因素......................................................................14
2-4  幾丁聚醣..................................................................................................16
2-4-1  幾丁聚醣之來源..........................................................................16
2-4-2  幾丁聚醣之性質..........................................................................17
2-4-3  電紡絲製備幾丁聚醣於組織工程應用......................................17
2-5  褐藻酸鈉..................................................................................................20
2-5-1  褐藻酸鈉之來源..........................................................................20
2-5-2  褐藻酸鈉之性質..........................................................................20
2-5-3  電紡絲製備褐藻酸鈉於組織工程應用......................................21
2-6  複合材料..................................................................................................22
2-6-1  複合材料之簡介..........................................................................22
2-6-2  複合材料於組織工程之應用......................................................23
2-7  電紡絲製備複合纖維..............................................................................25
2-8  電紡絲製備複合纖維於組織工程應用..................................................26
2-9  幾丁聚醣與褐藻酸鈉之複合材料..........................................................28
第三章  實驗..........................................................................................................32
3-1  實驗藥品..................................................................................................32
3-2  實驗儀器..................................................................................................33
3-3  實驗方法..................................................................................................35
3-3-1  電紡絲溶液之配製......................................................................35
3-3-2  纖維製備......................................................................................36 3-3-3  掃描式電子顯微鏡影像分析(SEM)...........................................36
3-3-4  黏度量測......................................................................................36
3-3-5  收集量量測..................................................................................37
3-3-6  混合纖維製備..............................................................................37
3-3-7  材料表面分析..............................................................................37
3-3-8  材料之生物適合性......................................................................39
第四章  結果與討論..............................................................................................41
4-1  電紡絲測試..............................................................................................41
4-1-1  Chitosan/PEO 之纖維型態測試..................................................42
4-1-2  Chitosan/PEO 溶液之黏度測量..................................................51
4-1-3  Alginate/PEO 之纖維型態測試 ..................................................54
4-1-4  Alginate/PEO 溶液之黏度測量 ..................................................63
4-2  chitosan/alginate 複合纖維製備 .............................................................66
4-2-1  纖維之收集量測試......................................................................66
4-2-2  纖維之螢光染色測定..................................................................69
4-2-3  X 射線光電子能譜儀測量..........................................................71
4-3  材料表面性質測定..................................................................................75
4-3-1  傅立葉轉換紅外線光譜分析儀(FT-IR) .....................................75
4-3-2  接觸角(contact angle)分析..........................................................78
4-4  生物適合性..............................................................................................80
4-4-1  纖維絲對細胞的貼附型態之行為..............................................80
4-4-2  細胞活性分析(MTT assay).........................................................85
4-4-3  乳酸脫氫酶(LDH )細胞毒性分析..............................................87
第五章  結論..........................................................................................................90
第六章  參考文獻..................................................................................................92

                                

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