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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃慶璨 | |
| dc.contributor.author | Jia-Lin Hong | en |
| dc.contributor.author | 洪佳麟 | zh_TW |
| dc.date.accessioned | 2021-06-14T17:09:43Z | - |
| dc.date.available | 2011-08-22 | |
| dc.date.copyright | 2011-08-22 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-12 | |
| dc.identifier.citation | 1. Ackroyd, R., C. Kelty, N. Brown, and M. Reed. 2001. The history of photodetection and photodynamic therapy. Photochem Photobiol 74:656-669.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40975 | - |
| dc.description.abstract | 生物膜 (biofilms) 為附著於物體表面生長的微生物群，可生長於醫療器材及傷口組織，形成持續性感染，對大部分殺菌劑有極強的抗性，因此不易治療。光動力治療 (photodynamic therapy) 為一新興的微生物感染治療方法，其殺菌能力不受細菌抗藥性影響而備受重視，已知對革蘭氏陽性之抗藥性菌株及生物膜具有殺菌效果，但對革蘭氏陰性菌的效果不佳，因此本研究探討以甲殼素 (chitosan) 促進光動力對革蘭氏陰性菌的殺菌作用。實驗結果發現亞甲基藍 (methylene blue) 光動力治療對革蘭氏陰性菌效果不彰，但經0.0004% 甲殼素處理10分鐘，光動力治療對革蘭氏陰性菌懸浮細胞的效果有顯著提升。在0.05% 甲殼素處理90分鐘，光照劑量480 J/cm2 的條件下，可對綠膿桿菌 (Pseudomonas aeruginosa) 生物膜完全殺菌。利用螢光光譜儀測定懸浮細胞的亞甲基藍激發量，發現激發量越高，殺菌效果越好，且當激發量超過約1 nmol/mL時可達完全殺菌，這與前人研究所發現光感物質激發量必須達到閾值 (threshold) 以上才能完全殺菌的現象一致。以甲殼素先處理，細菌的光感物質激發量顯著上升，這可說明甲殼素的確能幫助亞甲基藍吸附於菌體上。掃描式電子顯微鏡觀察發現經甲殼素處理後菌體會有聚集現象。雖甲殼素與光感物質的結合實驗無法完整解釋菌體吸附更多亞甲基藍的原因，但甲殼素合併光動力治療應是極具潛力的治療模式。 | zh_TW |
| dc.description.abstract | Biofilms are groups of microbes that attached to a surface. They can grow on indwell medical devices or human tissues, resulting in chronic or persistent infections. Because biofilms have stronger resistance to antibiotics than planktonic cells, those infections of biofilms are hard to treat. Photodynamic therapy (PDT) is an emerging method to treat microbial infections. So far, there is no strain resistant to PDT being observed. PDT is effective against Gram-positive planktonic cells and biofilms as well as drug-resistant strains. However, Gram-negative bacteria are more resistant to PDT. In this study, we used chitosan to enhance the efficacy of PDT. The results show that methylene blue-mediated PDT has a poor efficacy to Gram-negative bacteria, but it is more effective to planktonic cells treated with 0.0004% of chitosan for 10 minutes. Biofilms treated by 0.05% of chitosan for 90 minutes are also more sensitive to PDT. We observed that there is a threshold of methylene blue excitation level at 1 nmol/mL. If the excitation level of methylene blue is higher than the threshold, all the bacteria will be killed. After the treatment of chitosan, the amount of methylene blue binding to bacteria is increased, and the excitation level of methylene blue is higher. It implies that chitosan can enhance the binding between bacteria and methylene blue. The images taken by scanning electron microscope show bacteria aggregated after treatment of chitosan. Our result shows that chitosan have weak binding ability to methylene blue, it cannot explain the mechanism of chitosan enhancing PDT. However, the use of chitosan to enhance the efficacy of PDT may be a potent method. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-14T17:09:43Z (GMT). No. of bitstreams: 1 ntu-100-R98b47416-1.pdf: 10097689 bytes, checksum: 0f327426254b28d7c51131b49574219f (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 摘要 i
Abstract ii 目錄 iii 圖目錄 vi 壹. 前言 1 1、 微生物感染之防治 1 2、 生物膜 2 2.1. 環境中之生物膜 2 2.2. 生物膜之形成 2 2.3. 生物膜的結構 3 2.4. 生物膜之抗藥性機制 4 2.5. 生物膜對人類的害處 6 2.6. 生物膜防治 6 3、 光動力治療 7 3.1. 光動力治療之歷史 7 3.2. 光動力治療之機制 8 3.3. 光動力治療之要素 9 3.4. 亞甲基藍光動力治療 10 4、 甲殼素 (chitosan) 11 4.1. 甲殼素的特性 11 4.2. 甲殼素的應用 11 4.3. 甲殼素的抗菌機制 13 5、 研究動機及目的 14 貳. 材料與方法 16 1、 菌株 16 1.1. 懸浮菌體培養 16 2、 儀器 16 2.1. 光源 16 2.2. 生物膜培養系統 16 3、 藥品配製 19 3.1. 磷酸緩衝液 (PBS) 之配製 19 3.2. 亞甲基藍母液之配製 19 3.3. 甲殼素母液配製 19 4、 生菌數定量分析 19 5、 生物膜培養 20 6、 亞甲基藍對懸浮細胞之光動力治療 20 7、 亞甲基藍對生物膜之光動力治療 20 8、 亞甲基藍激發量分析 21 9、 雷射掃描共軛焦顯微鏡觀察 21 10、 甲殼素對懸浮細胞光動力治療之影響 21 11、 甲殼素對生物膜光動力治療之影響 22 12、 亞甲基藍與chitosan結合能力測試 22 13、 掃描式電子顯微鏡觀察 22 13.1. SEM觀察懸浮細胞 22 13.2. 蓋(載)玻片明膠覆膜 23 13.3. SEM觀察生物膜 23 13.4. 統計分析 23 參. 結果與討論 24 1、 亞甲基藍對懸浮細胞之光動力治療 24 1.1. 革蘭氏陽性菌 24 1.2. 革蘭氏陰性菌 27 2、 甲殼素合併光動力治療對革蘭氏陰性菌的殺菌效果 32 3、 亞甲基藍光動力治療於生物膜的殺菌效果 39 3.1. 生物膜培養與定量 39 3.2. 金黃色葡萄球菌生物膜累積曲線 39 3.3. 綠膿桿菌生物膜累積曲線 40 3.4. 亞甲基藍光動力治療於生物膜之殺菌效果 41 4、 甲殼素合併光動力治療對革蘭氏陰性菌生物膜的殺菌效果 44 5、 掃描式電子顯微鏡觀察菌體形態之變化 49 6、 以透析膜分析甲殼素與亞甲基藍的結合 54 7、 討論 56 7.1. 懸浮菌體光動力治療 56 7.2. 生物膜光動力治療 57 7.3. 透析膜實驗 57 肆. 結論 61 伍. 未來研究方針 62 陸. 參考文獻 63 柒. 附錄 69 | |
| dc.language.iso | zh-TW | |
| dc.subject | 光動力治療 | zh_TW |
| dc.subject | 生物膜 | zh_TW |
| dc.subject | 甲殼素 | zh_TW |
| dc.subject | 亞甲基藍 | zh_TW |
| dc.subject | methylene blue | en |
| dc.subject | chitosan | en |
| dc.subject | photodynamic therapy | en |
| dc.subject | biofilm | en |
| dc.title | 甲殼素對革蘭氏陰性菌懸浮細胞及生物膜亞甲基藍光動力殺菌效果之影響 | zh_TW |
| dc.title | Effects of chitosan on the efficacy of methylene blue mediated photodynamic therapy against Gram-negative bacterial planktonic cells and biofilms | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 許瑞祥,陳進庭,蔡翠敏,簡雄飛 | |
| dc.subject.keyword | 光動力治療,生物膜,亞甲基藍,甲殼素, | zh_TW |
| dc.subject.keyword | photodynamic therapy,biofilm,methylene blue,chitosan, | en |
| dc.relation.page | 72 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-12 | |
| 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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