溫感水凝膠及酵素部分融釋於骨韌帶癒合之研究

Thermosensitive Hydrogel and Partial Enzyme Digestion on Tendon-Bone Healing

黃裕閔(Yu-Min Huang)

指導教授 ： 林峯輝

共同指導教授 ： 曾永輝 ； 陳志華(Chih-Hwa Chen)

國立臺灣大學/工學院/醫學工程學研究所/博士(2020年)

https://doi.org/10.6342/NTU202002322

摘要

背景:前十字韌帶斷裂是膝蓋常見的疾病，當發生斷裂時會造成相當程度的功能喪失，手術重建前十字韌帶是目前治療的標準，然而韌帶重建的過程會有韌帶及骨頭癒合不良的問題，此一問題會造成復健的困難及影響手術的成功率，此研究是希望利用溫感水凝膠與酵素融釋的方式來解決骨韌帶癒合之問題，以期開發能臨床使用於前十字韌帶重建手術。
方法：將不同濃度的酵素及溫感水凝膠施行體外實驗，進行細胞毒性及酵素釋放效率測試後決定最適當之酵素濃度。接著進行兔子的動物實驗，取用兔子的長伸趾肌腱及脛骨長軸進行骨韌帶癒合之研究，利用36隻紐西蘭白兔的雙膝施作動物實驗，溫感水凝膠及酵素於手術時注射治療，術後進行組織切片檢查、斷層掃描、韌帶拉力測試來檢驗酵素融釋的效果。
結果與討論：酵素的體外實驗證實使用溫感水凝膠合併酵素可以有效促進韌帶的自我修復能力且無明顯細胞毒性，進行兔子動物實驗的切片檢查可見在四週及八週都有明顯的組織增生，且於八週時有明顯新生骨行成，斷層掃描可見於八週有顯著新生骨組織於酵素治療組，拉力測試可見酵素融釋可促進66%的最大荷重，證實使用溫感水凝膠合併酵素治療可有效促進骨韌帶癒合。
結論：溫感水凝膠與酵素融釋能成功促進骨韌帶癒合及新生骨的形成，並且提高生物力學的穩定性。這些結果表明溫感水凝膠與酵素融釋將可能使用前十字韌帶重建手術上。

關鍵字

溫感水凝膠 ； 酵素融釋 ； 骨韌帶癒合 ； 前十字韌帶 ； 組織工程

並列摘要

Background: Anterior cruciate ligament rupture is one of the most commonly injured structure in the knee joint, which induces severe comorbidity and functional deficits. Anterior cruciate ligament reconstruction is the gold standard treatment now. However, healing of an anterior cruciate ligament in bone tunnel yields weaker fibrous scar tissue, which may prolong the healing process within the tendon-bone interface. Poor tendon-bone healing may cause delay rehabilitation program and poor surgical results. In this study, we applied thermosensitive hydrogel with enzyme digestion to improve the healing process between tendon-bone interface.
Methods: In vitro study, we evaluated the different concentration enzyme to promote tendon graft healing. Cytotoxic test and enzyme release profile were also identified for the optimal enzyme concentration. New Zealand’s white rabbit’s long digital extensor tendon was detached and translated into a 2.5-mm diameter tibial plateau tunnel. Thirty-six rabbits underwent bilateral surgery and hydrogel injection with and without collagenase. After scarifying, we performed histological examination, computed tomography imaging, and biomechanical test to evaluate the effect of collagenase partial digestion on tendon-bone healing.
Results Discussion: In vitro study, better tendon healing was illustrated after collagenase partial digestion with thermosensitive hydrogel. Cytotoxic and collagenase release profile was also identified. In rabbit animal study, histological analyses revealed early healing and more bone formation at the tendon-bone interface after collagenase partial digestion. Micro computed tomography showed a significant increase in total bone volume and bone volume/tissue volume in the 8 weeks in the study group. Load-to-failure was significantly higher in the treated group at 8 weeks. Treatment with collagenase digestion resulted in a. 66% increase in pull-out-strength.
Conclusions: Injection of thermosensitive hydrogel with collagenase improves tendon-to-bone healing and more woven bone formation in a rabbit model. These results demonstrate that thermosensitive hydrogels with collagenase are potential candidates for clinical applications in anterior cruciate ligament reconstruction surgery.

並列關鍵字

thermosensitive hydrogel ； collagenase digestion ； tendon-bone healing ； anterior cruciate ligament ； tissue engineering

參考文獻

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