臨界尺寸骨缺損(critical-sized bone defect)的治療至今仍是臨床骨科中面臨的難題，由於大量的骨質流失，結構不穩定及缺血導致傷處產生大量的ROS及發炎反應，對整體癒合進程產生不良影響。一般以自體骨移植或同種異體骨移植作為兩種主要的治療方式，其仍有許多限制如:取得體積有限、具感染風險或不具骨誘導性等。在本研究中，我們將礦化膠原蛋白(mineralized collagen)結合矽奈米顆粒(silicon nanoparticles)製備為仿生骨組織工程支架，並利用控制溫度梯度製備出具有放射狀排列的纖維結構。本研究分為三部份，在材料方面，我們合成出的礦化膠原蛋白表現出近似生物骨的組成，並透過比較結構差異、機械強度、孔隙度及骨傳導性等條件找到最適合動物實驗之支架配方，而摻雜於支架內的矽奈米顆粒能夠在體外釋放具抗發炎能力的氫氣及具骨誘導性之矽酸。在細胞實驗中，含有矽奈米顆粒之支架具有降低免疫細胞內ROS及促炎細胞因子的水平、促進成骨細胞分化及誘導細胞遷移至支架中心等效果。在動物實驗中，我們將支架植入大於臨界尺寸之小鼠顱骨缺損內，證實此含有矽奈米顆粒之骨仿生支架能夠有效提高新骨生成比例，顯示其應用於臨界尺寸骨缺損再生的潛力。

Treatment of critical-sized bone defects is still a difficult problem in clinical orthopaedics due to massive bone loss. Structural instability and ischemia result in high ROS and inflammatory response in the defect area, which adversely affects the overall healing process. Autograft or allograft are the two choices of treatment, which still have many limitations, such as limited volume acquisition, risk of infection, or lack of osteoinductive properties. In this study, we prepared mineralized collagen combined with silicon nanoparticles as a biomimetic bone tissue engineering scaffold, under controlled temperature gradients, to produce a radially aligned fibrous scaffold. The as-synthesized mineralized collagen, which exhibited a composition similar to that of biological bone, and compared the structural differences, mechanical strength, porosity, and osteoconductivity to find the most suitable scaffold formulation for animal experiments, while the silicon nanoparicles incorporated in the scaffold were able to release anti-inflammatory hydrogen and osteoinductive silicic acid. In cellular experiments, the silicon nanoparticles-containing scaffold could effectively reduce the levels of ROS and pro-inflammatory cytokines in immune cells, promote osteoblast differentiation, and induce cell migration to the scaffold center. In an animal study, we implanted the scaffold into a mouse cranial defect larger than the critical size and demonstrated that the silicon nanoparticles-containing bone biomimetic scaffold was effective in increasing the rate of new bone formation, demonstrating its potential for the repair of critical-sized bone defects.

摘要 I
Abstract II
目錄 III
圖錄 V
表目錄 VII
第一章 緒論 1
1.1 臨界尺寸骨缺損 (critical-sized bone defect) 1
1.2 仿生骨組織工程支架 2
1.3 過量活性氧物質與骨缺損再生之關係 3
1.4 氫氣與抗發炎效果 4
1.5 矽應用於促進骨缺損再生之潛力 5
1.6 研究動機與實驗目的 7
第二章 材料與方法 10
2.1 支架製備 10
2.1.1 實驗材料 10
2.1.2 MCol製備方法 10
2.1.3 支架製備 10
2.2 MCol合成鑑定 11
2.2.1 傅立葉轉換紅外光譜(FTIR) 11
2.2.2 X光繞射儀(XRD) 11
2.2.3 同步熱分析儀(SDT) 11
2.2.4 元素分析 11
2.3 支架製備最佳化 12
2.3.1 MCol濃度最佳化 12
2.3.2 nSi摻雜量最佳化 13
2.4 nSi-MCol支架之釋放分析 13
2.4.1 氫氣釋放分析 13
2.4.2 離子釋放分析 13
2.5 細胞培養 13
2.6 nSi-MCol之生物毒性測試 14
2.7 nSi-MCol抑制細胞內ROS之定性評估 14
2.8 nSi-MCol抑制細胞內ROS之定量評估 15
2.9 nSi-MCol抑制促炎細胞因子分泌之定性評估 15
2.10 nSi-MCol抑制促炎細胞因子分泌之定量評估 15
2.11 nSi-MCol促進成骨細胞分化效果探討 16
2.11.1 成骨細胞增生 16
2.11.2 成骨細胞分化 16
2.12 nSi-MCol骨傳導性測試 16
2.13 細胞遷移實驗 17
2.14 動物實驗模型 17
2.15 成骨體積分析 18
2.16 組織染色切片 18
第三章 實驗結果與討論 19
3.1 支架特性分析 19
3.1.1 MCol合成結果探討 19
3.1.2 支架配方之最佳化 22
3.1.3 nSi-MCol之釋放性能分析 27
3.2 體外實驗 28
3.2.1 材料毒性測試 28
3.2.2 nSi-MCol之抗發炎效果 29
3.2.3 nSi-MCol對成骨細胞之影響 33
3.2.4 排列結構促進細胞遷移之效果 36
3.3 動物實驗 38
3.3.1 微型電腦斷層掃描 (𝜇CT)分析 38
3.3.2 組織切片染色分析 40
第四章 結論 42
參考文獻 43

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