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研究生： 林幸二
Lin, Sing-Er
論文名稱： 非熱微電漿系統之抑菌效用評估-使用細菌於模擬傷口之培養基模型
Bacterial inhibition effect for applying a non-thermal micro-plasma system:a proof-of-concept study with bacteria in a simulated wound-medium model
指導教授： 廖峻德
Liao, Jiunn-Der
共同指導教授: 劉浩志
Liu, Bernard Haochih
王士豪
Wang, Shyh-Hau
學位類別： 碩士
Master
系所名稱： 工學院 - 材料科學及工程學系
Department of Materials Science and Engineering
論文出版年： 2017
畢業學年度： 105
語文別： 中文
論文頁數： 68
中文關鍵詞： 非熱微電漿 、模擬傷口模型 、抑菌
外文關鍵詞： Non-thermal micro-plasma, simulated wound model, inhibition
相關次數： 點閱：123  下載：2 
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    • 電漿醫療器材於近幾年快速發展，本實驗室所開發的非熱微電漿裝置已通過國際電工委員會所設的醫療電氣設備的安全和有效性技術標準IEC 60601-1之電磁性安全規範，透過德國DIN工業標準局所設的DIN SPEC 91315法規與臨床動物安全試驗，驗證了此醫療器材之安全性，並通過了人體試驗委員會的審核標準，已可以應用在人體之上。以非熱微電漿系統應用於人體傷口之醫療器材，其功能性主要訴求改善傷口處微環境，提供一個適合傷口修復之環境，進而促使傷口癒合。遂本研究將使用本實驗室所開發之非熱微電漿裝置，觀察微電漿系統產生的抑菌圈大小與對於不同環境下之抑菌功效。使用菌種為臨床傷口上最常見之兩種菌種，金黃色葡萄球菌與綠膿桿菌。
    本研究先進行微電漿溫度與反應性物種診斷，實驗結果顯示微電漿系統於激發功率13 W，氮氣添加1%於氦氣中為最佳參數，依照診斷結果作為後續實驗參數。進行菌種生長曲線量測，找出菌種之活性較高，繁殖力也較強的生長時間點。實驗結果顯示金黃色葡萄球菌需生長3 hr，綠膿桿菌需生長4.5 hr。
    抑菌圈評估實驗，抑菌圈大小隨著處理時間的增加而增大。液相抑菌評估實驗，主要藉由反應性物種水解進入液相中，與水中分子與離子交互反應，產生氧化劑破壞液相中菌種之細菌膜，抑制水中菌種的生長。氣相抑菌評估實驗，以培養基模擬傷口溫床，模擬電漿於潔淨與惡劣微環境改善的實驗，電漿處理的組別與各別控制組相比，菌落數均有顯著差異的下降。其中對於潔淨微環境之環境改善程度較惡劣微環境的改善效果好。於細菌膜完整性評估實驗，經過電漿處理，其細菌膜皆有破損的情況產生，使染劑穿透破損細菌膜，於螢光顯微鏡下呈現鮮紅色。抑菌效果的有效性與生物膜的深度有關，可能與生物膜厚度成比例地下降。

    Plasma medical equipment evolve rapidly in recent years. The functional requirement of non-thermal micro-plasma system used on wound care is mainly to improve the wound micro-environment, to provide a suitable environment for wound repair, and thus promote wound healing. In this study, a non-thermal micro-plasma device is used to observe the size of the inhibition zone produced by the micro-plasma system and the inhibition effect for different in-vitro environments. The inhibition zone experimental results show that the size of the inhibition zone formed by the plasma increases with the increase in the treatment time. In liquid phase, the reactive species produced by micro-plasma system are hydrolyzed and produce oxidant to destroy the cell membrane of the bacteria and inhibit the growth of the bacteria. On the simulated wound model, the number of colonies of the test group significantly declined compared to the control group. Having two different bacteria cultivated on either clean micro-environment or harsh environment, after plasma treatment, the inhibition effect on the clean micro-environment was improved more significantly. The inhibition effect in the biofilm after the plasma treatment showed that the bacterial viability declined compared with the control group. The bacteria appeared to be in bright red under the microscope, due to the dye going through the damaged bacterial membrane. The effectiveness of the plasma is related to the depth of the biofilm and decreases with respect to the thickness of the biofilm.

    摘要 I Extended abstract II 誌謝 IX 目錄 XI 圖目錄 XIV 表目錄 XVIII 第一章 緒論 1 1.1 前言 1 1.2 研究動機 2 1.3 研究目的 6 第二章 理論基礎與文獻回顧 7 2.1一般傷口照顧 7 2.1.1 閉塞敷料(Occlusive dressing) 7 2.1.2 皮膚替代物(Skin substitutes) 7 2.1.3 超聲波(Ultrasound): 7 2.1.4 電療(Electrostimulation) 7 2.1.5 高壓氧治療(Hyperbaric oxygen therapy) 8 2.1.6 負壓傷口治療(Negative pressure wound therapy) 8 2.2 電漿 8 2.2.1 電漿簡介 8 2.2.2 大氣電漿 9 2.2.3 微電漿 9 2.3 電漿中的物種及效應 13 2.3.1 電漿物種與抑菌機轉 13 2.3.2 微電漿物種測量與診斷 16 2.4 微電漿應用於體外抑菌之評估 17 2.4.1 液相抑菌之評估 17 2.4.2 模擬傷口培養基抑菌之評估 19 2.4.3 生物膜上抑菌之評估 21 2.5 微電漿應用於細胞測試與傷口照顧 23 第三章 材料與方法 26 3.1 實驗設計與流程 26 3.2 非熱微電漿系統 27 3.3 細菌培養 28 3.3.1 培養基製作 28 3.3.2 菌種培養與菌種生長曲線 29 3.3.3 菌種計數 30 3.4 DIN法規-生物評估標準:微生物測試 31 3.5 細菌接種與體外測試 32 3.6 抑菌存活率評估 34 3.7 細菌螢光影像 34 3.8 分析儀器 35 3.8.1 螢光顯微溫度計 35 3.8.2 光學放射光譜儀 36 3.8.3 螢光顯微鏡 36 第四章 微電漿系統診斷與菌種生長曲線 37 4.1 電漿溫度與物種量測 37 4.2 微電漿系統之特性光譜分析 38 4.2.1 電漿全區掃描分析 38 4.2.2 電漿特定物種之光譜掃描與半定量分析 39 4.3 菌種生長曲線量測 43 4.4 第四章整理 44 第五章 微電漿系統之體外抑菌測試 45 5.1 DIN法規測定-生物評估標準:微生物測試 45 5.1.1 抑菌圈大小評估 45 5.1.2 液相抑菌效果評估 47 5.2 模擬傷口與微環境改善評估 48 5.2.1 潔淨微環境 48 5.2.2 惡劣微環境 50 5.3 細菌膜完整性評估 53 5.4 第五章整理 57 結論 58 未來展望 59 參考文獻 60

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