利用細胞效應與滅菌效率評估大氣低溫平面電漿對傷口癒合之影響

非熱介電層阻擋放電電漿（DBD）目前已廣泛於醫療應用上，為新穎有效傷口治療工具，可促進細胞增殖、細胞遷移與傷口癒合，且可對活體組織進行滅菌，成為具有潛力的工具。低劑量的活性氧物種透過生長因子相關機制，可增加內皮細胞的遷移、增生及血管形成；反之，高劑量活性氧物種會造成細胞傷害甚至凋亡。傷口癒合時期，發炎期須避免受細菌感染，而纖維母細胞在增生期扮演著重要角色。因此本研究使用由實驗室共同開發之大氣低溫平面式電漿系統，此系統反應器經出氣改良後可在一大氣壓下產生電漿，且證實對細胞有高敏感性以及具滅菌效果之活性氧物種，利用光學放射光譜儀(OES)分析得知本系統在波長777、845、927 nm觀察到單態氧及297、309 nm觀察到氫氧自由基等物種。在連續激發電漿600秒時處理溫度低於34℃，不會對組織或傷口造成熱傷害。利用0~300秒時間參數進行電漿處理評估纖維母細胞移動能力，經培養36小時觀察，在電漿處理300秒以下可使纖維母細胞的細胞移動速率上升，其中以30、60秒移動距離最佳，相較於控制組移動距離增加51.2、50.2 μm。本研究利用此系統對大腸桿菌及金黃色葡萄球菌之滅菌效率評估，經電漿分別處理300秒（大腸桿菌）與480秒（金黃色葡萄球菌）滅菌程度可達D-value，處理時間300秒以上（大腸桿菌）及540秒以上（金黃色葡萄球菌）可達完全滅菌。綜合以上研究結果，依照傷口癒合時期訂定使用建議，於發炎期時搭配傷口滅菌效果以電漿治療300秒，可達大腸桿菌99.99%、金黃色葡萄球菌70.15%滅菌效果，纖維母細胞較控制組活性下降25%、細胞移動距離增加19.32%；增生期搭配電漿治療60秒，與控制組相比纖維母細胞活性較提高50%，細胞移動距離增加42.11%，大腸桿菌與金黃色葡萄球菌則有44.58%及28.07%滅菌效果。在傷口癒合發炎期與增生期搭配不同療程將促進傷口癒合達到最優化。

關鍵字

大氣低溫電漿；纖維母細胞；傷口癒合；電漿滅菌

並列摘要

The applications of non-thermal dielectric barrier discharge plasma (DBD) have been used widely in biomedical area nowadays. Throughout the wound healing period, the wound should avoid to any bacterial infection during the inflammation stage and the fibroblast plays an important role in the proliferation phase. Therefore, the study is to develop a non-thermal atmospheric planar plasma system. The ameliorate of the nozzle on the planar reactor can produce plasma under atmospheric pressure and it has been proved to have high sensitivity to the cells with high sterilization effect. Using the optical emission spectroscopy, the singlet oxygen can be detected under 777, 845 and 927 nm wavelength and 297 and 309 nm for free radical species. In 600 s and 34ºC excitation of plasma, it will not cause any thermal damage to tissues or wound. While the migration rate of the fibroblast is found to be best under 300s of treatment throughout 36 h observation on the cultivation. Among them, 30s and 60s are the best parameters for the period of treatment, which increase of 51.2 and 50.2 µm cell movement distance respectively. The study also showed on the inactivation of E. coli and S. aureus can achieve D-value under 300s and 480s respectively and complete sterilization when the time is over 300s and 540s for E. coli and S. aureus respectively. In comprehensive of the result, it is suggested to treat the wound with plasma during inflammation phase for 300s, in order to achieve 99.99% of E. coli and 70.15% of S. aureus sterilization effect, but the activity of fibroblast will be decreased by 25%, migration rate increased by 19.32%. In 60s of treatment during proliferative phase, the activity of the fibroblast is increased by 50%, migration rate increased by 42.11%, but the sterilization effect on the E. coli and S. aureus can only achieve 44.58% and 28.07% respectively. It is recommended to use suggested parameters matching with inflammation phase and proliferative phase for the best effect on wound healing.