臺灣博碩士論文加值系統

褥瘡是由於組織長時間的受到壓迫，導致該部位的微血管阻塞，沒有足夠的血液輸送氧氣，使細胞死亡速度加快，進而導致皮膚及組織壞死、潰瘍的現象。目前對於褥瘡傷口感染的治療仍然是將抗生素大量投入人體中，然而在未正常使用下往往會產生具抗藥性的細菌出現，導致傷口感染治療難度提升。本研究提出運用光動力控制系統抑制傷口分離菌之構想，利用光動力療法對細菌作用機轉與抗生素治療相異，降低細菌發生抗藥性的機率，選擇並設計適合大面積貼附於傷口表面之光源，藉由控制系統穩定光源能量輸出，提升患者治療之便利性與穩定性，再搭配分次照光模式克服褥瘡傷口組織缺氧的問題。本研究針對褥瘡傷口常見之具有抗藥性與非抗藥性金黃色葡萄球菌，使用光感藥物靛花青(Indocyanine green, ICG)以不同照光模式、光照能量、及光感藥物濃度進行系統性光動力殺菌驗證。
本研究為了使光源能夠盡量貼附於人體表面，光源溫度應必須考量，因此選擇在光功率密度為5 mW/cm2下，光源表面溫度42 oC，並以光譜儀驗證LED發光波長為 770 ± 5 nm，符合光感藥物ICG之吸收峰值。本研究自製之光照控制系統能夠在分次照光下，提供穩定光功率照設。由光動力殺菌實驗結果顯示，以低光功率密度5 mW/cm2進行光動力治療時，光照能量在25 J的效果優於其他能量實驗組；藥物濃度在12.5 g/ml的效果較其他藥物濃度實驗組佳；分次照光模式殺菌效果也較連續照光實驗組效果佳。本研究藉由自製之光動力控制系統試驗體外殺菌實驗結果可以顯示出在傷口分離菌上確實有滅菌成效，並具有褥瘡傷口臨床應用之潛能。

Pressure sores are usually caused by impaired blood supply and tissue malnutrition owing to prolonged pressure over bony prominences. Pressure sores are difficult to treat due to wound infection and the development of drug resistant bacteria. This study proposed the photodynamic therapy (PDT) control system to inactivate bacteria isolated from the wound. The bacteria killing mechanism of PDT is different from antibiotics and thus can reduce the development of drug resistance. We designed the light source so that it can be attached to the skin and used fractionated illumination mode to solve the problem of wound hypoxia. We used two bacterial strains which are common pathogens on pressure sores: oxacillin-resistant Staphylococcus aureus (ORSA) and oxacillin-susceptible Staphylococcus aureus (OSSA). Indocyanine green (ICG) was used as photosensitizer.
This system consists of LED light source which emits 770 ± 5 nm wavelength of light. The absorption peak of ICG was confirmed with a spectroscope. Considering light source will apply to irradiate closely to patient. The temperature on light source surface was 42 oC with irradiance was at 5 mW/cm2. The results showed that the PDT control system can provide stable irradiance on fractionated illumination mode. The antibacterial effects in the energy of 25 J and concentration of 12.5 μg/ml were found to be more effective than other parameters. Moreover, the fractionated illuminated mode provided better results than continuous illumination mode. In conclusion, through in vitro study results, the implemented PDT control system has potential to inactivate bacterial infections of pressure sores and is a good reference for future clinical application.

目錄

摘要 I
Abstract II
謝誌 III
目錄 IV
圖目錄 VII
表目錄 IX
第一章 緒論 1
1.1 研究背景 1
1.2 文獻回顧 2
1.2.1 光動力治療歷史 2
1.2.2 光動力治療殺菌歷史 3
1.2.3 光動力治療於金黃色葡萄球菌之成效 4
1.2.4 分次與低光功率密度累積之光動力治療應用 4
1.2.5 靛花青(indocyanine green, ICG) 5
1.3 研究動機與目的 6
1.4 論文架構 7
第二章 理論基礎 8
2.1 微生物簡介 8
2.2 抗藥模式與機轉 9
2.3 光動力治療 9
2.3.1 光動力治療機制 9
2.3.2 光感藥物 11
2.3.3 氧氣 12
2.3.4 光源 12
第三章 研究方法與材料 14
3.1 褥瘡光動力治療系統設計 15
3.2 顯示介面與按鍵控制設計 16
3.3 系統驗證 17
3.3.1 光源波長測試 17
3.3.2 光源光功率及溫度測試 17
3.3.3 光動力控制系統功能驗證 17
3.4 微生物滅菌實驗 18
3.4.1 菌種介紹 18
3.4.2 微生物初始濃度配置 18
3.4.3 光動力治療滅菌實驗設計 19
第四章 研究結果 22
4.1 系統驗證 22
4.1.1 光源溫度及光功率測試 22
4.1.2 光動力治療波長驗證 24
4.1.3 光動力控制系統功能驗證 24
4.1.4 顯示介面呈現 25
4.2 光動力殺菌實驗結果 26
4.2.1 累積照光能量12.5 J/cm2結果 27
4.2.2 累積照光能25 J/cm2結果 29
4.2.3 累積照光能31.25 J/cm2結果 31
4.2.4 溶氧實驗結果 33
4.2.5 氧氣控制細菌實驗結果 33
第五章 討論 35
5.1 光源光功率密度與溫度探討 35
5.2 不同藥物濃度與累積光照能量殺菌效果探討 36
5.3 連續照光與分次照光殺菌效果探討 39
5.4 OSSA及ORSA之光動力治療差異探討 40
第六章 結論與未來展望 41
6.1 結論 41
6.2 未來展望 42
參考文獻 43
附錄 A 實驗試劑及使用儀器 49
附錄 B 實驗藥物配置 50

圖目錄
圖1- 1研究目的示意圖 6
圖2- 1光動力治療反應機制示意圖 10
圖2- 2 ICG吸收鋒值，最大吸收峰波長775 nm 11
圖3- 1研究架構 14
圖3- 2控制裝置區塊圖架構 15
圖3- 3控制裝置實照圖 16
圖3- 4 LED陣列光源實照圖 16
圖3- 5 系統運作示意圖 16
圖3- 6連續光照實驗設計圖 19
圖3- 7分次光照實驗設計圖 20
圖4- 1光源表面溫度量測結果 23
圖4- 2光源背面溫度量測結果 23
圖4- 3光功率量測結果 23
圖4- 4LED波長測試結果 24
圖4- 5控制系統穩定度測試結果 25
圖4- 6介面顯示結果 25
圖4- 7殺菌實驗示意圖 26
圖4- 8 連續照光模式，累積光照強度12.5 J/cm2 28
圖4- 9分次照光模式，累積光照強度12.5 J/cm2 28
圖4- 10連續照光模式，累積光照強度25 J/cm2 30
圖4- 11連續照光模式，累積光照強度25 J/cm2 30
圖4- 12連續照光模式，累積光照強度31.25 J/cm2 32
圖4- 13連續照光模式，累積光照強度31.25 J/cm2 32
圖4- 14　 LB broth培養液通氧實驗結果 33
圖4- 15 ORSA 氧氣控制細菌實驗結果 34
圖4- 16 OSSA 氧氣控制細菌實驗結果 34
圖5- 1不同濃度ICG吸收波長量測結果 36
圖5- 2 不同濃度ICG於比色管中顏色差異 38
圖5- 3 兩組光源進行光動力治療示意圖 38
圖5- 4 兩組光源進行光動力治療實驗結果 38

表目錄
表2- 1常見用於光動力治療之光源特性 13
表3- 1各種細菌稀釋溶液與吸光值 18
表5- 1 ORSA兩種照光模式統計分析結果 39
表5- 2 OSSA兩種照光模式統計分析結果 39
表5- 3 連續照光模式ORSA與OSSA統計分析結果 40
表5- 4分次照光模式ORSA與OSSA統計分析結果 40

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