- 學位論文
血小板裂解液與高能聚焦超音波對軟骨再生治療之應用
Applications of Platelet Lysate and High Intensity Focused Ultrasound in Treatment of Cartilage Regeneration
摘要
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。
並列摘要
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.