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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳進庭(Chin-Tin Chen) | |
| dc.contributor.author | Chueh-Pin Chen | en |
| dc.contributor.author | 陳玨頻 | zh_TW |
| dc.date.accessioned | 2021-06-08T02:13:57Z | - |
| dc.date.copyright | 2015-12-23 | |
| dc.date.issued | 2015 | |
| dc.date.submitted | 2015-12-18 | |
| dc.identifier.citation | 1. 何漣漪. 奧妙無窮的微生物世界, 科學發展, 2003, 361, 48-53.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/19696 | - |
| dc.description.abstract | 自然界中的微生物大部份以生物膜形式存在，生物膜對殺菌劑有極強之抗性。此外，抗生素的廣泛及不當使用導致抗藥性菌株的產生。而這些抗藥性菌株的出現，讓許多研究學者開始尋求其他治療的方式。光動力殺菌為一新興的微生物感染治療方式，研究結果顯示這一個殺菌方式並不會對微生物造成抗藥性。然而光動力殺菌目前於動物實驗及臨床使用上仍存在許多問題，像是對革蘭氏陰性菌及生物膜殺菌效果不如預期。先前研究發現，利用幾丁聚醣結合光動力治療，光動力能夠對菌體造成破壞、殺死菌體，再加入幾丁聚醣可增強殺菌之作用，降低藥物的使用量，進而提高殺菌之效果。於臨床應用上，如果藥物劑型流動性高，不易附著於感染部位，將導致治療效果不彰。為因應光動力殺菌實際需求，須找出一最佳治療之配方，希望可長時間附著於感染部位上，且不會影響藥物釋放。本研究中，首先將開發一應用於治療微生物感染之幾丁聚醣水凝膠搭配光動力殺菌技術平台，並利用牙周治療來確認其效益。實驗中，以HPMC為增稠劑來提高幾丁聚醣水凝膠黏度，隨著HPMC濃度增加，幾丁聚醣水凝膠黏度及黏附力皆會增加。將幾丁聚醣水凝膠與金黃色葡萄球菌及綠膿桿菌生物膜共同培養後，進行照光，於照光後持續培養，殺菌效果隨幾丁聚醣水凝膠黏度增加，有下降之趨勢，且由共軛焦顯微鏡觀察，TBO釋放受到HPMC濃度之影響，故培養時間長短影響光動力效益。動物實驗上，使用幾丁聚醣水凝膠可減少感染傷口之菌數。接著，建立一體外牙齦槽模型並於模型中測試幾丁聚醣水凝膠之光動力效益，發現增加照光劑量及培養時間，會增加光動力殺菌之效果；照光的數目對殺菌效果也有顯著的影響。以牙周病致病菌株作測試，亦有同樣的殺菌效果。其次，探討光動力對生物膜之作用機制，發現光動力除造成菌體死亡外，亦會破壞生物膜之胞外聚合物，使幾丁聚醣能進一步達到協同殺菌之效果。最後，基於光動力處理會破壞生物膜之機制，於光動力處理後加入抗生素，發現不需提高藥物濃度即可增加殺菌效果，且此效果不僅針對標準菌株生物膜，對抗藥性菌株之生物膜亦有相同效果。 | zh_TW |
| dc.description.abstract | Antibiotics are the most common antimicrobial agents used for the treatment of bacteria and fungi infection. Drug-resistance is a growing problem largely due to the widespread use of antibiotics. Biofilms are the main growth forms of microbes in nature and have stronger resistance to antibiotics compared to planktonic cells. Photodynamic inactivation (PDI) is an emerging method to treat microbial infections. Presently, there is no report related microbial resistance to PDI. However, there are several problems to be resolved for PDI application. Previously, we showed that chitosan could increase the efficacy of PDI against both Gram-positive and Gram-negative bacteria in planktonic cells and biofilms. The purpose of this study was to develop a chitosan hydrogel containing hydroxypropyl methylcellulose (HPMC), chitosan and toluidine blue O (TBO) to improve the PDI efficacy for topical application in clinic. The PDI efficacy of hydrogel was examined in vitro against the biofilms of Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa). Confocal scanning laser microscopy (CSLM) was performed to investigate the penetration level of TBO into viable S. aureus biofilms. Incorporation of HMPC could increase the physicochemical properties of chitosan hydrogel including the hardness, viscosity as well as bioadhesion; however, higher HMPC concentration also resulted in reduced antimicrobial effect. CSLM analysis further demonstrated that higher HPMC concentration constrained TBO diffusion into the biofilm. The bactericidal efficacy could be significantly augmented by prolonged retention of hydrogel in the biofilm as well as in the infected skin burn wounds of rat after light irradiation. We found that increasing the illumination dose and incubation time could enhance the PDI efficacy. The PDI efficacy of chitosan hydrogel was also verified in the biofilm of periodontal pathogenic strains in a 3D gingival model. Further studies indicate that PDI could damage the extracellular polymer substrate of biofilm. Finally, we showed that combination of PDI and antibiotics could significantly increase the bactericidal effect against biofilm of wild type strains as well as drug-resistant strains. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T02:13:57Z (GMT). No. of bitstreams: 1 ntu-104-D00b22004-1.pdf: 6014925 bytes, checksum: 5f438140a479e5c5b5ea6078e328a90a (MD5) Previous issue date: 2015 | en |
| dc.description.tableofcontents | 中文摘要 I
Abstract II 目錄 III 圖表目錄 VII 附圖表目錄 IX 第一章、緒論 1 1-1 微生物與人類 1 1-1.1 微生物的感染防治 1 1-1.2 常見的微生物感染 2 1-1.3 生物膜 2 1-1.4 抗藥性機制 4 1-2 抗生素 6 1-2.1 抗生素的發展 6 1-2.2 抗生素之治療與瓶頸 7 1-3 光動力作用 8 1-3.1 光動力之歷史及應用 8 1-3.2 光動力治療之作用機轉 8 1-3.3 參與光動力治療的主要因子 9 1-4 光動力殺菌之發展與限制 11 1-5 光動力的應用 13 1-5.1 細菌及真菌in vitro 研究 13 1-5.2 皮膚感染 13 1-5.3 牙周病之治療 13 1-6 幾丁聚醣(Chitosan) 14 1-6.1 幾丁聚醣抗菌模式 15 1-6.2 幾丁聚醣的抗菌效能 16 1-6.3 幾丁聚醣之應用 16 1-6.4 幾丁聚醣於光動力之應用 17 1-7 幾丁聚醣水凝膠(Hydrogel) 17 1-8 本研究菌種介紹 18 第二章、研究動機與目的 20 第三章、實驗架構 21 第四章、實驗材料與方法 22 4-1 菌種來源與保存 22 4-2 實驗材料與儀器 23 4-3 實驗方法 26 4-3.1 幾丁聚醣水凝膠的製備 26 4-3.2 黏度分析(Viscosity) 26 4-3.3 質地分析(Texture Profile Analysis, TPA) 26 4-3.4 可注射性分析(Assessment of the Injectability) 27 4-3.5 懸浮菌體培養方法 27 4-3.6 生物膜培養 27 4-3.7 雷射掃描共軛焦顯微鏡(Confocal Scanning Laser Microscopy, CSLM) 27 4-3.8 懸浮菌體之光動力殺菌 28 4-3.9 光動力於生物膜之殺菌 29 4-3.10 市售抗生素凝膠於生物膜之殺菌 29 4-3.11 光動力殺菌in vivo動物實驗 30 4-3.12 建立牙齦槽模型 31 4-3.13 黏附力研究 31 4-3.14 牙齦槽模型之生物膜培養 31 4-3.15 抗生素配製 32 4-3.16 抗生素敏感性分析 32 4-3.17 抗生素於生物膜上之殺菌 32 4-3.18 光動力結合抗生素殺菌實驗 32 4-3.19 統計分析 33 第五章、結果 34 【第一部分：幾丁聚醣水凝膠開發及應用】 34 5-1 幾丁聚醣水凝膠配方設計 34 5-2 幾丁聚醣水凝膠之特性分析 34 5-3 幾丁聚醣水凝膠可注射性分析 35 5-4 幾丁聚醣水凝膠對金黃色葡萄球菌生物膜之光動力效益 35 5-5 幾丁聚醣水凝膠於金黃色葡萄球菌生物膜中TBO滲透實驗 36 5-6 持續培養對幾丁聚醣水凝膠於金黃色葡萄球菌生物膜之光動力效益 37 5-7 幾丁聚醣水凝膠對綠膿桿菌生物膜之光動力效益 38 5-8 持續培養對幾丁聚醣水凝膠於綠膿桿菌生物膜之光動力效益 38 5-9 傾斜角度對光動力之影響 38 5-10 大鼠感染傷口測試 39 【第二部分：牙齦槽模型建立及幾丁聚醣水凝膠應用於牙周病治療】 40 5-11 牙齦槽模型之建立 40 5-12 幾丁聚醣水凝膠之黏膜黏附力 40 5-13 幾丁聚醣水凝膠於牙齦槽中金黃色葡萄球菌生物膜之光動力殺菌效益 41 5-14 照光劑量及照光數量對牙齦槽中金黃色葡萄球菌生物膜之光動力殺菌效益 41 5-15 幾丁聚醣水凝膠對牙齦槽中牙周病菌株之光動力殺菌效益 42 【第三部分：光動力與幾丁聚醣對生物膜作用機制】 43 5-16 由掃描式雷射共軛焦顯微鏡觀察生物膜之活菌死菌 43 5-17 由掃描式雷射共軛焦顯微鏡觀察生物膜之胞外聚合物 44 【第四部分：光動力結合抗生素於生物膜殺菌效果之探討】 45 5-18 抗生素對懸浮菌體之敏感性分析 45 5-19 抗生素對生物膜之殺菌效益 46 5-20 抗生素增強對生物膜光動力殺菌之效益 46 5-21 抗生素對抗藥性懸浮菌體之敏感性分析 47 5-22 抗生素對抗藥性菌株生物膜之殺菌效益 47 5-23 抗生素增強對抗藥性菌株生物膜光動力殺菌之效益 47 第六章、討論 49 6-1 幾丁聚醣抗菌模式 49 6-2 幾丁聚醣水凝膠之材料篩選 50 6-3 幾丁聚醣水凝膠 51 6-4 幾丁聚醣水凝膠對生物膜之光動力殺菌探討 53 6-5 傾斜角度對光動力殺菌之影響 54 6-6 幾丁聚醣水凝膠於in vivo 測試 54 6-7 幾丁聚醣水凝膠於牙齦槽模型之光動力殺菌探討 55 6-8 齒槽模型之應用性 57 6-9 市售水凝膠與幾丁聚醣水凝膠 58 6-10 光動力作用對生物膜之影響 58 6-11 光動力增強抗生素的殺菌效果之探討 59 第七章、結論 61 圖 62 表 101 附圖表 109 參考文獻 122 | |
| dc.language.iso | zh-TW | |
| dc.title | 光動力殺菌幾丁聚醣水凝膠之開發及齒槽模型建置 | zh_TW |
| dc.title | Developments of Chitosan Hydrogel and Dental Gingival Model for Antimicrobial Photodynamic Inactivation | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 蔡翠敏,楊正昌,林晉玄,謝堅銘 | |
| dc.subject.keyword | 光動力殺菌,生物膜,幾丁聚醣,水凝膠,抗生素, | zh_TW |
| dc.subject.keyword | photodynamic inactivation,biofilm,chitosan,hydrogel,antibiotic, | en |
| dc.relation.page | 131 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2015-12-18 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科技學系 | zh_TW |
| 顯示於系所單位： | 生化科技學系 | |
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