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研究生:林恕安
研究生(外文):Shu-An Lin
論文名稱:血小板裂解液與高能聚焦超音波對軟骨再生治療之應用
論文名稱(外文):Applications of Platelet Lysate and High Intensity Focused Ultrasound in Treatment of Cartilage Regeneration
指導教授:郭柏齡郭柏齡引用關係李百祺
指導教授(外文):Po-Ling KuoPai-Chi Li
口試委員:沈哲州廖愛禾
口試委員(外文):Chen-Chou ShenAi-Ho Liao
口試日期:2017-11-16
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:生醫電子與資訊學研究所
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2018
畢業學年度:106
語文別:中文
論文頁數:60
中文關鍵詞:退化性關節炎膝蓋軟骨Safranin-OMasson基質侵入式脈衝式高能聚焦超音波血小板裂解液注射治療超音波影像發炎反應恢復週期
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退化性關節炎是運動或勞動中常見的傷害因素。目前臨床治療主要僅為減緩受損或退化速度,而再生治療為目前新興的治療方式,但其處裡的程序尚未被確立。因此,本研究主要的目的在於確立一個有效的軟骨重建流程,藉由針與高能量聚焦超音波直接對膝蓋軟骨造成破壞,再利用血小板裂解液注射療法於紐西蘭白兔膝蓋軟骨,使其再生。
第一部分為破壞實驗。首先,我們利用針戳以及高能聚焦超音波 (High intensity focused ultrasound, HIFU)對膝蓋軟骨造成傷害,其中,藉由HIFU產生之傷害另外以超音波B-mode影像之斑塊訊號的變化進行確認膝蓋軟骨基質 (matrix)是否有微破壞之產生,最後再針對膝蓋軟骨中的Safranin-O及Masson染色上做比較,進而評估破壞之程度。
第二部分為軟骨血小板裂解液注射療法 (platelet lysate, PL)之驗證。在確認膝蓋軟骨傷害之效果後,我們比較了細胞自行修復以及配合血小板裂解液注射療法之兩種情況。本部分透過4隻紐西蘭公兔 (每隻約3公斤)之活體實驗驗證,內容包含探討配合血小板裂解液治療之療效、是否有機會減短軟骨再生所需之時間。同時,我們透過超音波系統抓取特徵值,利用三角面積計算方式,抓取軟骨的厚度,對未來的長期追蹤實驗預作準備。
本研究亦規劃在未來將脈衝式高能聚焦超音波應用於治療的可能性。我們計畫在活體實驗中加上脈衝式高能聚焦超音波造成的傷害與復原程度之比較,藉由每周定位再生之軟骨厚度來追蹤回復情況,同時比較各種回復狀況,最後確立血小板裂解液注射療法在活體實驗中的效果。
Osteoarthritis (OA) is commonly seen and responsible for substantial morbidity both in sports and in the workplace. In general, the purpose to disease treatment is to recover original activity. Regeneration therapy is new method right now, but still not effective treatment program in clinical. We use needle trauma and HIFU (High intensity focused ultrasound) to cause damage to cartilage, and use PL (Platelet lysate) treatment to treat in New Zealand rabbit’s articular cartilage to cause regeneration.
First part, we demonstrated the pulsed-HIFU can significantly cause damage articular cartilage in ex-vivo as indicated by the change echogenicity in ultrasound B-mode images and results in the Safranin-O and Masson stains to the damage of extracellular matrix. Second part, we use New Zealand rabbit for each one is 3 kg, 3 months, and use 24G needle to cause damage, to compare with needle trauma and needle trauma with PL treatment to prove that PL treatment can reduce cartilage recovery cycle and cartilage regeneration. Otherwise, we use ultrasound to detect the depth of cartilage by used triangle area method to auto catch the cartilage in image and calculate the depth of cartilage to prepare long-term follow-up experiment in future.
In the future, we plan the possibility of applying pulsed-HIFU to therapy. We plan to compare the extent of injury and recovery with pulsed-HIFU in vivo and track recovery by locating the regenerated cartilage thickness weekly, and also comparing status of recovery. Finally, To establish PL therapy effect in vivo experiment.
口試委員會審定書 #
誌謝 i
中文摘要 ii
ABSTRACT iii
目錄 iv
圖目錄 vii
表目錄 xi
Chapter 1 緒論 1
1.1 研究動機 1
1.2 膝關節退化和治療之介紹 1
1.2.1 軟骨結構之介紹 1
1.2.2 膝關節退化 2
1.2.3 膝關節疾病治療方法 3
1.2.4 高濃度血漿血小板注射治療 3
1.2.5 血小板裂解液注射治療 4
1.3 臨床前研究 5
1.3.1 手術誘導膝蓋關節炎模型 5
1.3.2 注射藥劑誘導膝蓋關節炎模型 6
1.3.3 針戳及高能聚焦超音波造成膝蓋關節炎模型 6
1.4 研究目標 7
1.5 論文架構 7
Chapter 2 實驗原理與方法 8
2.1 高能聚焦超音波夾具製作 9
2.1.1 製具設計原理 9
2.1.2 製具共焦點 10
2.2 活體實驗架構 13
2.2.1 血小板裂解液實驗及針戳破壞 13
2.2.2 超音波量測軟骨厚度 18
Chapter 3 結果與討論 20
3.1 兔子針戳破壞結果 20
3.2 離體豬膝蓋軟骨於高能聚焦超音波破壞性結果 21
3.3 活體實驗 25
3.4 討論 34
3.4.1 離體豬膝蓋軟骨於高能聚焦超音波破壞性 34
3.4.2 活體實驗 34
Chapter 4 結論與未來工作 35
4.1 結論 35
4.2 未來展望 35
Chapter 5 附錄 36
5.1 附錄一:治療性高能量聚焦超音波探頭製造與測量 36
5.1.1 研究動機 36
5.1.2 超音波探頭基本結構 36
 背膠層 38
5.1.3 研究目標 39
5.1.4 實驗方法與量測 39
5.1.5 結果 48
5.1.6 討論 49
5.1.7 結論 50
5.2 附錄二:超音波彈性影像對肌腱之應用於紐西蘭白兔 50
5.2.1 研究目標 50
5.2.2 肌腱疾病和修復之介紹 50
5.2.3 超音波影像基礎理論 51
5.2.4 靜態彈性影像 52
5.2.5 動態彈性影像 52
5.2.6 傳統剪切波彈性陣列架構 53
5.2.7 超聲速彈性影像 54
5.2.8 實驗方法 55
5.2.9 結果與討論 55
 結果 55
 討論 56
 結論 56
REFERENCE 57
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