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研究生:李志鴻
研究生(外文):Chih-Hung Lee
論文名稱:將多孔海綿狀Chitosan固定於PET不織布/PNIPAAm水膠複合表面上製備三層創傷敷料
論文名稱(外文):Porous Chitosan Sponge Immobilized onto the Surface of PET Nonwoven/ PNIPAAm Hydrogel Composites to Fabricate Tri-layer Wound Dressing
指導教授:陳克紹陳克紹引用關係
指導教授(外文):Ko-Shao Chen
學位類別:碩士
校院名稱:大同大學
系所名稱:材料工程學系(所)
學門:工程學門
學類:綜合工程學類
論文種類:學術論文
論文出版年:2005
畢業學年度:93
語文別:英文
論文頁數:61
中文關鍵詞:支架細胞培養創傷敷料幾丁聚醣
外文關鍵詞:cell culturechitosanscaffoldwound dressing
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在這研究中,是將具有生物分解性以及抑菌性幾丁聚醣高分子固定在PET不織布/PNIPAAm水膠複合表面上應用於創傷敷料。PET不織布被廣泛的使用是因為具有多孔性、高表面積以及強大的機械性質。然而,PET不織布的表面疏水性是應用上的一個限制。所以我們使用電漿活化處理以及UV光接枝聚合NIPAAm水膠來改善PET不織布的適應性。
結果顯示,PET不織布經過電漿處理以及光接枝處理後是有改善其濕潤性。NIPAAm是有特別的熱感應性質。易剝離表面是由NIPAAm分子固定在PET不織布所決定的,在PNIPAAm水膠/PET不織布複合物表面固定幾丁聚醣,再以冷凍乾燥方式以及三磷酸鈉交聯反應製備出多孔海綿狀幾丁聚醣三層創傷敷料。
分析將有FT-IR以及SEM。孔洞結構可由SEM觀察到。多孔海綿狀幾丁聚醣在類似體內模式經過30天後,殘餘率還有76%,因此,多孔海綿狀幾丁聚醣為生物分解性材料,且分解率不會太快,抑菌能力也會顯現。在細胞培養中,細胞可以生長在多孔海綿狀幾丁聚醣,而老鼠胚胎幹細胞生長在經過Ar處理是沒這麼好的,而在多孔海綿狀幾丁聚醣添加過膠原蛋白以及組織培養皿的生長曲線是類似的。幾丁聚醣三層敷料使老鼠在12天後傷口愈合,且無排斥發炎現象。
Chitosan sponge, which is a biodegradable and antibacterial polymer immobilized onto the surface of PNIPAAm gel/Poly (ethylene terephthalate) (PET) non-woven composites surface for application in wound dressing was studied. Poly (ethylene terephthalate) non-woven has extensively using due to its porosity, allowing ventilation, high surface area and excellent mechanical properties. However, the hydrophobic surface of PET non-woven had limited on its application, so, we tried to use the plasma-activation treatment and subsequently UV-light graft polymerization of NIPAAm gel to improving the compatibility of PET non-woven with PNIPAAm gel. The results indicated that the wet ability was improved after plasma treatment and photo-induced graft. Graft monomer was N-isopropylacrylamide (NIPAAm), which has specific thermo-sensitive property. The substrate of a easy strip surface was prepared by introduction of NIPAAm monomer onto PET non-woven, these complexes were characterized by prepared chitosan porous scaffold, finally, chitosan was immobilized onto PNIPAAm gel/PET non-woven composites surface using freeze-dried treatment and the cross-linking reagent, Na5P3O10. The designed products were analyzed by FT-IR and scanning electron microscopy (SEM). Porous structure was shown in SEM pictures. Chitosan sponge in vivo likely status for 30 days and chitosan sponge remain rate was 76%. So chitosan sponge was bio-degradation material, and degradation rate wasn’t too fast. Antibacterial activities will be executed. For cell culture, cells can grow in chitosan sponge, and mouse embryonic stem cells grew in chitosan sponge that treatment by Ar plasma was not well. Then, mouse embryonic stem cells grew in chitosan sponge and chitosan sponge that with 10μl type I collage and tissue culture plate were similar. In animal test, chitosan tri-layer wound dressing make mice wound healed after 12 days, and no inflammation status. And it can be expected to applicative in wound dressing.
Contens
Chinese abstract…………………………………………...………...…Ⅰ
English abstract………………………………………………...…...…II
Contents………………………………………………………………..IV
List of Figures………………………………………………………....ⅥI
List of Tables……………………………………………………..….....IX
1. Introduction………………………………………………………….1
2. Literature Review…………………………………………………….…………..5
2.1.1 Chitin……………………………………………………...…..5
2.1.2 Chitosan……………………………………………………….5
2.1.3 Chitosan as biomedical membranes…………………………...6
2.1.4 Role of chitosan in drug delivery systems………………….…7
2.1.5 Chitosan as an artificial skin…………………………………..7
2.2.1 Hydrogels……………………………………………………….8
2.2.2 PNIPAAm-Thermal-sensitive polymer…………………………9
2.3 Poly (ethylene terephthalate)…………………………………….10

3. Experiment
3.1Materials…………………...……………………………………….11
3.2Plasma treatment………………..………………………………….11
3.3Photo-induced graft copolymerization………...…………………...12
3.4.1Preparation of chitosan solution………………………………….12
3.4.2Preparation of chitosan spong……………………………………12
3.4.3 Preparation of easily stripped chitosan wound dressing………...13
3.5.1 FTIR analysis……………………………………………………13
3.5.2 Morphological observation……………………………………...13
3.5.3 Water content……………………………………………………14
3.5.4 Gel content………………………………………………………14
3.5.5 Water vaporization transmission rate……………………………14
3.5.6 Degradation rate…………………………………………………15
3.6.1 Cell culture………………………………………………………15
3.6.2 MTT assay………………………………………………………16
3.6.3 Histological examination………………………………………..16
3.6.4 Assay for antibacterial activity………………………………….17
3.7 In vivo evaluation-animal test…………………………………….17
3.7.1 Histological examination………………………………………..18

4. Results and Discussion
4.1PNIPAAm hydrogel observation………...…………………………19
4.2Tri-stripped wound dressing observation……...…………………...19
4.3.1 FTIR analysis……………………………………………………19
4.3.2 Morphological observation……………………………………...20
4.3.3 Water content……………………………………………………21
4.3.4 Gel content………………………………………………………21
4.3.5 Water vaporization transmission rate……………………………22
4.3.6 Degradation rate…………………………………………………22
4.4.1 Cell proliferation and differentiation……………………………23
4.4.2 Morphology of stem cells cultured on chitosan scaffold……….23
4.4.3 Histological observation of cell culture…………………………24
4.5 Bacteriostasis activity……………………………………………..24
4.6.1 In vivo wound healing experiments……………………………..25
4.6.2 Histological observation of animal test………………………….25

5. Conclusions…………………………………………………………27

6. Reference……………………………………………………………50



List of Figures
Figure 1-1 Structure of skin…………………………………………...29
Figure 1-2 The skin area of body……………………………………...29
Figure 1-3 Schematic diagram of a bilaminar artificial……………….29
Figure 1-4 Diagram of Burke and Yannas artificial skin……………...29
Figure 2-1 Structure of chitin and chitosan…………………………...29
Figure 4-1 OM of thermal-sensitive PNIPAAm gel at temperature…..30
Figure 4-2 OM of tri-layer easily stripped wound dressing…………..31
Figure 4-3 Fourier transform infrared spectra of chitosan, chitosan cross-linked Na5P3O10, and Na5P3O10…………...………...32
Figure 4-4 SEM of freeze-dried and crosslinked chitosan sponge……33
Figure 4-5 Water vaporization transmission rate of chitosan sponge and PET non-woven and control………………………...…….36
Figure 4-6 Degradation remain rate of chitosan sponge crosslinked by Na5P3O10 and non-crosslink…………...…………………..37
Figure 4-7 OM of mouse embryonic stem cells in chitosan sponge….38
Figure 4-8 MTT of STO cells attached on tissue culture pate………...39
Figure 4-9 MTT of embryonic stem cells attached on tissue culture pate……………………………………………………...…40
Figure 4-10 MTT of STO cells attached on chitosan sponge…………41
Figure 4-11 MTT of embryonic stem cells attached on chitosan sponge……………...……………………………………...42
Figure 4-12 Histological of mouse embryonic stem cells attached on chitosan sponge……………………………………………43
Figure 4-13 SEM of mouse E. S. cells attached on chitosan sponge…44
Figure 4-14 SEM of mouse E. S. cells attached on chitosan sponge…45
Figure 4-15 The method of prevention of bacteria's further growth and the results of prevention of bacteria's further growth .…. 46
Figure 4-16 OM of animal test………………………………………..47
Figure 4-17 Histological of animal test……………………………….48
Figure 4-18 OM of amimal test2……………………………………...49




List of Tables
Table 1. Water content for chitosan sponge……………………………..34
Table 2. The effect of concentrations of crosslinking agent and chitosan on gel content……………………...……………………………35
Reference
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