||Efficacy study of non-thermal micro-plasma for promoting wound healing
||Department of Materials Science and Engineering
皮膚上有許多問題如皺紋、疤痕、色素沉澱與粉刺等問題，可經由雷射手術將上述問題獲得改善。經雷射照射後的傷口需時四至十二周可完成皮膚重塑，而在傷口癒合的照護上使用aquaphor healing ointment、biafine toptical emulsion、polysporin、neosporin等常見臨床用治療藥膏，但部分藥膏有刺激性，使用時會伴隨刺痛感，且藥物過敏者不建議使用，且使用藥膏需每天更換敷藥。所以提出一種非接觸式、無痛、不造成過敏並具抗菌附加效果，且免除每天頻繁換藥的治療方式。故使用非熱微電漿系統在雷射照射後傷口進行治療，探討其表皮與真皮介面接合與促進傷口癒合。
非熱微電漿系統以射頻電源供應器作為激發源，工作距離固定為4 mm。主要激發氣體為氬氣與氮氣，流量分別為5 slm與5 sccm。在此工作距離下調控電漿激發功率與氮氣及氬氣混合比例進行電漿物種與溫度診斷。因此以激發功率為17 W時，電漿溫度低於37℃；並以氮氣混合比例為0.1%時，NO物種之相對強度為純氬氣電漿之八倍為治療參數。主要在非熱微電漿中的物種為NO、OH、O、Ar等。實驗分為三組：未治療組、電漿治療一次組、電漿治療三次組(術後前三天)。以傷口面積評估法評估癒合速率。癒合品質部分，以光學同調斷層掃描(OCT)、免疫組織化學染色與血流分布造影評估。傷口面積評估顯示，電漿治療一次後平均十八天結痂掉落，較未治療組平均二十五天結痂掉落縮短28% 的癒合時程。OCT與組織化學染色觀察中，電漿治療三次組在術後第七天觀察到表皮與真皮介面接合，即上皮再生。而電漿治療一次與三次組在術後七天也觀察到發展良好的肉芽組織。而未治療組在術後第十四天才出現表皮與真皮介面接合。OCT影像積分密度量化，在術後第十四天電漿治療三次組上升87.2%，電漿治療一次組上升87.6%，未治療組上升49.6%。血流通量分布造影部分，在術後第二十一天電漿治療組三次組上升60.4%，電漿治療一次組上升35.2%，未治療組上升37.0%。由縮短癒合時程、OCT影像積分密度與血流通量分佈，顯示電漿治療在促進傷口癒合與表皮與真皮介面接合之成效。
Currently, wound healing and skin regenerations are one of the major public health concerns which required considerable method and takes time. The laser irradiation is the frequently using method for the facial problems such as wrinkle, scar, pigmentation and acne. In addition, it could be promotes the remodeling of dermal-epidermal junction and need 4~12 weeks for complete recovery from wound healing and skin regeneration. In general, the common clinical prescription of the laser irradiation of the wound has been used by healing ointment such as aqurphor healing ointment (AHO), biafine topical emulsion (BTE), polysporin and neosporin, etc. However, healing ointment has might feel uncomfortable, tingling and allergy problem for some patients which indicates the urgent need to develop effective techniques to overcome the above problems. In the recent years, plasma therapy has attracted widespread interest which providing more advantage such as contact-free, painless, non-allergy and with antibacterial effect.
In this present work, we are introducing non-thermal micro-plasma techniques which significantly inducing skin remodeling dermal-epidermal junction and promoting wound healing. Herein, the animal modeling was divided into three conditions such as non-treatment (NT), plasma treatment once (PT1) and plasma treatment three times (PT3). The non-thermal micro-plasma was generated from radio frequency and working distance between skin surface and micro-plasma was 4 mm. The optimized micro-plasma plume temperature 37°C was controlled by adjusting applied power and nitrogen gas insert. The plasma species was investigated with optical emission spectroscopy (OES) which resulted that the NO, OH, O, and Ar were major species in the non-thermal micro-plasma system. The reduction percentage of wound area at the 21 postoperative days (POD) was found to be 80%, 90% and 100% for NT, PT1 and PT3 respectively in presence of nitrogen 0.1%.
In addition, the highest intensity of NO species was significantly increased by the laser irradiated wound healing. The wound healing quality assessments were studied by the optical coherence tomography (OCT), histochemistry and immunohistochemistry staining. The remodeling of dermal-epidermal junction, reepithelialization and well-developed granulation tissues were observed at 7 POD for PT1, PT3 group and 14 POD for NT group. The quantification of OCT image intensity at 14 POD and blood flow intensity at 21 POD results suggest that NT, PT1, PT3 group increased 49.6%, 87.6%, 87.2% and 37.0%, 35.2%, 60.4% by comparing with 1 POD intensity. Theses evidences proved that this novel non-thermal micro-plasma technique promising to be used as a fast and effective treatment for laser irradiated wound healing and remodeling the dermal-epidermal junction.
第一章 序論 1
1.1 前言 1
1.2 研究動機 1
1.3 文獻回顧 2
1.3.1 微電漿形式 3
1.3.2 非熱微電漿在動物模式上傷口癒合的可適用性 5
1.3.3 以非熱微電漿產生一氧化氮分子對傷口癒合之影響 10
1.3.4 非熱微電漿應用在雷射照射後傷口癒合 11
1.4 研究目的 13
第二章 理論基礎 14
2.1 雷射與皮膚醫學 14
2.2 傷口癒合 17
2.3 電漿 18
2.3.1 電漿簡介 18
2.3.2 大氣電漿 20
2.3.3 微電漿 21
2.3.4 電漿中的物種與效應 25
第三章 材料與方法 29
3.1 實驗設計與流程 29
3.2 實驗材料與製備 30
3.2.1 實驗動物 30
3.2.2 傷口製造 30
3.2.3 非熱微電漿系統 31
3.3 分析儀器 34
3.3.1 光學放射光譜儀 34
3.3.2 螢光式光纖溫度計 37
3.3.3 雷射都卜勒血流分佈造影 37
3.3.4 光學同調斷層掃描 39
3.3.5 組織病理學分析 40
第四章 非熱微電漿系統診斷與雷射照射傷口模式的建立 44
4.1 非熱微電漿系統溫度量測 44
4.2 非熱微電漿系統之特性光譜解析- 光學放射光譜儀 46
4.2.1 電漿全區段光譜掃描分析 46
4.2.2 電漿中特定物種之光譜掃描與半定量分析 48
4.3 二氧化碳雷射強度與照射後傷口深度關係 52
第五章 非熱微電漿系統在傷口癒合速率、品質與組織學的評估 55
5.1 傷口癒合速率 - 傷口面積評估法 55
5.2 傷口癒合品質評估 58
5.2.1 血流分布評估 58
5.2.2. 光學同調斷層掃描 61
5.3 傷口在組織學的評估 65
5.3.1 組織化學染色- H&E stain 65
5.3.2 免疫組織化學染色- 層黏蛋白(Laminin) 67
5.3.3 免疫組織化學染色- 基質金屬蛋白酵素第三型 (MMP-3) 69
5.4 非熱微電漿對重塑表皮與真皮介面接合與促進雷射照射傷口癒合之可能機轉 71
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