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論文中文名稱:幾丁聚醣與褐藻酸鈉其交聯物製成之多孔性支架緩慢釋放已酮可可鹼
之抗發炎效果 [以論文名稱查詢館藏系統]
論文英文名稱:The Effects of Controlled Release of Pentoxifylline from Crosslinked Chitosan and Alginate Porous Scaffolds in Suppressing Inflammatory Reactions [以論文名稱查詢館藏系統]
院校名稱:臺北科技大學
學院名稱:工程學院
系所名稱:生物科技研究所
畢業學年度:97
出版年度:98
中文姓名:邱欣洋
英文姓名:Sin-Yang Ciou
研究生學號:96688012
學位類別:碩士
語文別:中文
口試日期:2009-07-20
論文頁數:86
指導教授中文名:林忻怡
指導教授英文名:Hsin-Yi Lin
口試委員中文名:黃聲東;蔡偉博
口試委員英文名:Sheng-Tung Huang;Wei-Bor Tsai
中文關鍵詞:幾丁聚醣褐藻酸藥物釋放抗發炎
英文關鍵詞:ChitosanAlginateDrug releaseAnti-inflammatory
論文中文摘要:本研究主要在利用幾丁聚醣 ( CS ) 與褐藻酸鈉 ( A ) 研發出適合用於緩慢傳送藥物的多孔性支架並與抗發炎藥物 ( 已酮可可鹼 ) 結合,並探討抗發炎之功效。將交聯形成之複合高分子製成四組多孔性支架:1.幾丁聚醣 ( CS );2.幾丁聚醣與褐藻酸鈉交聯 ( CSA );3.幾丁聚醣、褐藻酸鈉經氯化鈣交聯( CSACa );4.褐藻酸鈉經氯化鈣交聯 ( ACa )。製作出之支架再進行物理性質測試與體外細胞抗發炎之實驗。由紅外線光譜觀察得知,CS的胺基與A上羧酸基交聯產生離子吸引,形成混合型吸收波峰,A中的羧酸基因受到鈣離子吸引,使的羧酸基吸收峰轉移到較高波長,鈣離子同樣會影響CSA之交聯物,均為有效交聯,再以掃描式電子顯微鏡觀察發現,經交聯的支架可使孔洞結構更為緊密;因此在壓縮結果分析,CSACa交聯複合物大幅提高抗壓強度並可減少降解程度。此外,在接觸角結果顯示,CS與A交聯後親水性增加,但CSA經氯化鈣交聯後較原支架疏水性提高;由於CS經交聯後對多孔性支架結構及親水性的改變,交聯後支架膨潤率較低。其上述因素皆會影響藥物釋放之程度,結果顯示,CSACa支架釋放藥物的速率顯著的較其他三組低。因此,幾丁聚醣與褐藻酸鈉多孔性支架確實可藉由交聯方式,有效延長藥物輸送的時間。另外,在細胞相容性實驗中顯示,幾丁聚糖與褐藻酸鈉其交聯物支架都具備有良好的細胞適應性;經LPS刺激RAW264.7細胞與多孔支架緩慢釋放PTX共同培養,測試其分泌的TNF-α及IL-6作為細胞發炎反應的指標。結果證實經培養6及24小時後,支架緩慢釋放PTX皆可達到較佳的抗發炎效果,CSACa釋放之PTX為四組中最持久,其抑制發炎因子效果較另三組為佳,可有效增強抑制發炎反應。
論文英文摘要:The purpose of this research was to develop a crosslinked chitosan ( CS ) and aliginate ( A ) porous scaffold that can be suitable for controlled drug release. Four materials made porous scaffolds were 1. chitosan ( CS ) , 2. chitosan crosslinked with alginate ( CSA ) , 3. chitosan crosslinked with alginate and Ca2+ ( CSACa ) , and 4. alginate crosslinked with Ca2+ ( ACa ). Esperiments performed on the crosslinked materials and scaffolds that physical properties were examined and in vitro effects of anti-inflammatory. The FTIR results showed Amide bonds were formed between CS and A because of the ionic attraction between NH3+ and COO-. Also, the ionic bonding between calcium ion and the alginate carbonyl group shifted the carboxyl peak to higher wavenumber. The calcium ion also effectively bonded to the COO- and improved the CSA mixture crosslinking. SEM showed CS crosslinked with A scaffold had smaller pores and more compact structure. CS hybrid scaffolds had stronger mechanical property ( higher Young’s modulus ) and less susceptible to enzymatic degradation than CS scaffolds. The hydrophilicity of CS increased after it was crosslinked with alginate. However, after crosslinked with Ca2+ the hydrophilicity of CSA decreased. Scaffolds made of CS crosslinked with A that change scaffold mechanical and hydrophilicity had less swell ratio than CS scaffolds. The release rate of PTX of CSACa slowest compared with the others groups. When the four groups of scaffolds were loaded with PTX and cultured with lipopolysaccharide-activated macrophages, the CSACa group had the best results in suppressing the release of inflammatory factors ( TNF-α and IL-6 ) after 6 and 24 hours. The slow PTX release from the CSACa scaffolds was thought to be the reason that improved the anti-inflammatory effect of PTX. We have successfully developed a tissue engineered scaffold that could not only used in controlled release but also has more effectively suppress inflammation.
論文目次:中文摘要 i
英文摘要 iii
誌謝 v
目錄 vi
表目錄 ix
圖目錄 x
第一章 前言 1
第二章 文獻回顧 3
2.1組織工程 3
2.2幾丁質與幾丁聚醣之介紹 5
2.3褐藻酸鈉 9
2.4幾丁聚醣與褐藻酸鈉交聯材料 11
2.5冷凍乾燥法………………………………………………………...13
2.6免疫系統與發炎…………………………………………………….14
2.7 巨噬細胞 15
2.8 脂多醣體 16
2.9抗發炎藥物Pentoxifylline簡介 17
第三章 實驗方法、藥品與儀器 19
3.1實驗材料 19
3.1.1細胞來源……………………………………………………....19
3.1.2儀器設備…………………………………………………… …19
3.1.3實驗藥品……………………………………………………….20
3.2幾丁聚醣與其交聯物薄膜、支架的製備…………………………...23
3.2.1幾丁聚醣與褐藻酸鈉其交聯物分組表與配製…………… ….23
3.3實驗設計與架構……………………………………………………...26
3.3.1幾丁聚醣與褐藻酸鈉其交聯物物理性質測試架構…………..26
3.3.2掃描式電子顯微鏡……………………………………………..27
3.3.3傅立葉轉換紅外線光譜儀……………………………………..27
3.3.4接觸角測試…………………………..........................................27
3.3.5壓縮測試………………………………………………………..28
3.3.6酵素降解測試…………………………………………………..29
3.3.7膨潤率…………………………………………………………..29
3.3.8藥物釋放測試…………………………………………………..30
3.3.9幾丁聚醣與其交聯物細胞測試架構與方法…………………..31
3.3.10 RAW264.7 巨噬細胞株之培養……………………………....32
3.3.11 RAW264.7 巨噬細胞與支架相容性測試……………………32
3.3.12 RAW264.7 巨噬細胞抗發炎測試……………………………33
3.3.13細胞活性………………………………………………………35
3.3.14細胞數………………………………………………………....35
3.3.15細胞激素之測定……………………………………………….36
3.3.15.1腫瘤壞死因子………………………………………….36
3.3.15.2介白素6………………………………………………..36
3.4統計分析……………………………………………………………...37
第四章 結果與討論…………………………………………………………….38
4.1幾丁聚醣與褐藻酸鈉其交聯物物理性質分析………………………38
4.1.1傅立葉轉換紅外線光譜分析…………………………………..38
4.1.2幾丁聚醣與褐藻酸鈉其交聯物外觀…………………………...42
4.1.3幾丁聚醣與褐藻酸鈉其交聯物以SEM觀察…………………43
4.1.4接觸角測試…………………………………………………….47
4.1.5膨潤率測試…………………………………………………….49
4.1.6壓縮測試……………………………………………………….51
4.1.7酵素降解測試………………………………………………….53
4.1.8藥物釋放分析………………………………………………….55
4.2幾丁聚醣與褐藻酸鈉其交聯物細胞測試……………………………57
4.2.1支架對RAW 264.7 細胞的相容性分析……………………...57
4.2.2支架緩慢釋放藥物對RAW 264.7 細胞抗發炎之影響……...62
第五章 結論………………………………………………………………….74
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