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研究生：	林秉佳 Lin, Ping-Chia
論文名稱：	透明質酸以組織工程方式修復老鼠阿基里斯腱斷裂模型之評估 Evaluation Hyaluronic Acid Tissue Engineering Repairs for Achilles Tendon Injury in Rat Model
指導教授：	葉明龍 Yeh, Ming-Long
學位類別：	碩士 Master
系所名稱：	工學院 - 醫學工程研究所 Institute of Biomedical Engineering
論文出版年：	2011
畢業學年度：	99
語文別：	英文
論文頁數：	81
中文關鍵詞：	阿基里斯肌腱、Modified Bunnell縫合技術、肌腱細胞、透明質酸、肌腱癒合
外文關鍵詞：	Achilles tendon, Modified Bunnell method, Tenocyte, Hyaluronic acid, Tendon healing
相關次數：	點閱：183 下載：1
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阿基里斯腱是全身最大的肌腱，在腳踝連接跟骨與小腿肌肉，由腓腸肌和比目魚肌的肌腱部分共同組成，其主要功能為將肌肉收縮所產生的力量傳送到骨頭，為人體最容易發生斷裂的肌腱之一。根據先前研究肌腱受傷後提供modified Bunnell縫合的固定方法，可以在早期提供較好的復原狀況肌腱受傷後提供一個穩定的固定方法，可以在早期提供較好的復原狀況。然而有許多研究顯示在肌腱斷裂處其細胞凋亡的速率會比正常情況下來的快上許多，而肌腱細胞正是提供分泌肌腱細胞外基質的主要成分。因此本實驗目的為以modified Bunnel為基礎縫合固定，再以組織工程方式將細胞透過載體放置於傷口處，期望能有更好的結果。實驗所選用之載體為透明質酸天然高分子，其存在於許多動物體內，且有良好之生物相容性。
本實驗使用大鼠為動物模型，將肌腱橫切後以Modified Bunnell縫法進行手術，接著再分成透明質酸/肌腱細胞、透明質酸以及生理食鹽水處理三個組別在術後癒合第四、七、十四、二十一天使用功能性評估以及力學測試為依據來評量。
實驗結果顯示Modified Bunnell縫法為一穩定的縫合方式，無論是在哪個組別其所形成的肌腱間間隙皆沒有明顯差異。而在力學強度與功能性上可發現各組別皆有隨著時間有所增加。值得注意的是經由透明質酸/肌腱細胞組的處理後無論是在力學、徑度、功能性評估以及踝關節活動度上都可以發現到在術後十四天即能恢復到術後二十一天的現象，顯示以這樣的方式對於肌腱癒合早期能有較佳的癒合效果。

Achilles tendon is a connective tissue composed of parallel array of collagen fiber and is the largest tendon in the body. It is one of tendons which suffer rupture most frequently, and the common rupture location is 2-7 cm above calcaneus. According to previous studies, Achilles tendon rupture repaired with suture could provide excellent fixation and functional recovery in early stage, especially using the modified Bunnell suture method that provides with the best early mechanical recovery. However, some studies have shown higher cell apoptosis in the position tendon rupture site than normal conditions. The structure and function such as secretion and synthesis of extracellular matrix of tendon are regulated mainly by the tenocytes. Hyaluronic acid (HA) is a natural biomaterial abundant in several animal tissues and present with good cell compatibility. The purpose of this study is based on tissue engineering concept to combine tenocytes with HA gel to enhance the sutured repair ruptured Achilles tendon in rat model.
The rat Achilles tendon was cut with blade and fixed with the modified Bunnell suture method. A total of 120 rats were used and were randomly assigned into three groups: normal saline (NS), hyaluronic acid (HA) and hyaluronic acid + tenocyte (HAT) groups respectively. Rats were sacrificed at 4, 7, 14 and 21 days after surgery and Achilles tendon functional assessment by gait analysis, healing strength and stiffness by mechanical testing, histology by HE stain were evaluated consequently. Observed mechanical and functional properties in each group had increased significantly over time. And noteworthy that all outcomes at 14 days in HAT group do not show any difference with 21 days, indicating the tendon might have reached full recovery at 14 days in HAT group. The results of this study suggested that using Modified Bunnell suture combining injection of hyaluronic acid mixture with tenocytes to repair the rat Achilles tendon injury had better results.

中文摘要	I
Abstract	III
致謝	V
Table of Contents	IX
List of Figures	XII
List of Tables	XVI
Chapter I Introduction	1
1 Achilles tendon structure and mechanical properties	1
2 Achilles tendon rupture	3
3 Tendon healing	4
4 Treatment of Achilles tendon ruptures	5
4.1 Surgical treatment	5
4.2 Traditional treatment	7
5 Hyaluronic acid	8
6 Literature	10
6.1 Functional assessment	10
6.2 Using different suture methods for repair of tendon mechanical properties	13
6.3 The influence of tendon gap formation on tendon healing	14
7 Rationale	15
8 Purpose	16
Chapter II Material and methods	17
1 Experimental equipment and materials	17
1.1 Material testing machine	17
1.2 Image acquisition system of tensile test	18
1.3 Fixture of tensile test	18
1.4 Gait functional evaluation system	19
1.5 Suture and suture methods	19
1.6 Experimental animals - rats	21
2 Experimental methods	21
2.1 Experimental Design	21
2.2 Surgical procedures	23
2.3 Tenocyte isolation	25
2.4 Preparation of material	26
2.5 Functional evaluation	26
2.6 Biomechanical test	29
2.7 Histological examination	31
2.8 Statistical analysis	32
Chapter III Results	33
1 Cell biocompatibility between tenocyte and HA	33
2 Postoperative anatomical appearance of the Achilles tendon	34
3 Postoperative tendon gap	35
4 Postoperative tensile strength	37
5 Postoperative stiffness	41
6 Postoperative functional evaluation	46
6.1 AFI	46
6.2 ROM	49
7 Histological analysis	60
8 Integration results	63
Chapter IV Discussion	66
1 Basic characteristics of rat Achilles tendon	66
2 Tenocytes Compatibility	66
3 Surgical suture and tendon gap	67
4 Sample storage and processing	68
5 Mechanical evaluation	68
6 Functional evaluation	70
7 Comparison with non-injection group	71
8 Limitations	72
9 Future researches	73
Chapter V Conclusion	74
Chapter VI References	75
個人基本資料與求學經歷	80

                                    

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