臺灣博碩士論文加值系統

本研究目的是開發一個具藥物釋放功能之可注射性鈣矽複合骨水泥，以促進骨組織修復。使用明膠包覆中藥所形成的微粒與鈣矽組成之粉體結合成一複合材料。另外，本實驗中也探討加入抗生素後對於發炎反應的作用。我們針對此鈣矽複合骨水泥之藥物釋放、注射性、浸泡前後的徑向拉伸強度、型態、相組成以及生物性質作了一系列的測試。結果顯示加入明膠微粒之後會延長藥物釋放的時間，在注射性方面，單純的鈣矽骨水泥在五分鐘後只剩下不到40%可注射性，而包含有微粒的組別二十分鐘後還保有約一半的可注射性，三十分鐘後剩下20%。在細胞培養的部分，包含有續斷的微粒增進細胞的生長比單純微粒的高出1.35倍，而包含骨碎補的組別則高出約1.2倍。在基因表現上，此鈣矽複合材料具備有好的成骨特性，亦有許多對於發展牙科與骨科材料所應具備的優點，因此我們認為在骨科修復方面，此材料是具有發展潛力的。

The aim of this study was to develop novel injectabile calcium silicate-based composite bone cements with drug release for bone repair. The gelatin microspheres containing Chinese herbs were incorporated into calcium silicate powders to form the composite. Moreover, we investigated the effect of the inflammation after adding antibiotics. The drug release, injectability, before and after immersion behavior in physiological solution, including diametral tensile strength, morphology, and phase composition of various cements were evaluated, in addition to biology properties. The results indicated that the gelatin micropaticles may prolong drug release time. The results of injectability show that the pure calcium silicate cement only remained 20% after 5 minutes, however, the composite containing microspheres can extend injection time until 30 minutes. After 20 minutes, the composite containing microspheres can remain 40% injectability. This biocompatibility study indicated the composite containing drug significantly enhanced cell proliferation and differentiation. After Xu-Duan release in the medium could promote cell viability and have 1.35 times higher than the group without drug. Gu-Sui-Bu had about 1.2 times than pure microspheres. Gene expression showed these calcium silicate composites with high osteogenecity. It is concluded that the composite cement are a potential material for bone defect repair.

目錄
致謝 II
中文摘要 III
Abstract IV
圖目錄 IX
第一章 文獻回顧及理論基礎 1
1-1前言 1
1-2 生醫骨科材料的種類及發展 1
1-2-1 金屬材料 2
1-2-2 高分子材料 2
1-2-3 陶瓷材料 3
1-2-4 複合材料 3
1-3 骨水泥 4
1-4 天然高分子在生醫材料的應用 7
1-4-1 明膠(gelatin) 7
1-5 藥物 8
1-5-1骨碎補( Gu-Sui-Bu ) 9
1-5-2 續斷( Xu-Duan ) 9
1-5-3 抗生素(gentamicin) 9
1-6 研究動機與目的 9
第二章 材料與方法 11
2-1 材料製備 11
2-1-1 鈣矽粉體製備 11
2-1-2 微粒製備 11
2-2 物理性質分析 12
2-2-1 硬化時間測試 12
2-2-1 浸泡實驗 12
2-2-2-1 成分分析 12
2-2-2-2 徑向拉伸強度 12
2-2-2-3 重量損失 13
2-2-2-4 掃描式電子顯微鏡觀察 13
2-2-3 可注射性 13
2-2-4 藥物釋放測試 13
2-3 生物性質分析 14
2-3-1 不同藥物濃度的細胞培養 14
2-3-2 微粒釋放的細胞培養 14
2-3-3 直接培養 14
2-3-4 Ca2+染色 15
2-3-5 掃描式電子顯微鏡觀察 15
2-3-6 光學顯微鏡觀察細胞 15
2-3-7 基因測試 16
2-3-7-1 RNA萃取 16
2-3-7-2 RT-PCR放大 16
2-3-7-3 Agarose gel 16
第三章 結果與討論 17
3-1 物理性質分析 17
3-1-1微粒表面形態觀察 17
3-1-2 浸泡實驗 18
3-1-2-1 X光繞射分析 18
3-1-2-2 徑向拉伸強度 22
3-1-2-3 重量損失 24
3-1-2-4 掃描式電子顯微鏡觀察 25
3-1-3 可注射性 28
3-1-4 藥物釋放測試 29
3-2 生物性質分析 32
3-2-1 不同藥物濃度的細胞培養 32
3-2-2 微粒釋放的細胞培養 34
3-2-3 直接培養 35
3-2-4 Ca2+染色 36
3-2-5 掃描式電子顯微鏡觀察 37
3-2-6 基因表現 40
3-2-6-1 發炎反應 40
3-2-6-2 成骨作用 43
第四章 結論 46
第五章 參考文獻 47

圖目錄
Fig. 1 經交聯後之微球表面型態 17
Fig. 2 不同骨水泥浸泡於SBF後所得之X光繞射分析結果 21
Fig. 3 骨水泥含不同藥物浸泡於SBF後所得之逕向拉伸強度 (a) 骨碎補 (b) 續斷 (c)gentamicin 23
Fig. 4 骨水泥浸泡於Tris buffer後之重量損失變化 24
Fig. 5 將不同骨水泥浸泡入(a) SBF中1天 (b) Tris buffer 7天後表面及斷面的顯微結構圖 27
Fig. 6 含有不同藥物骨水泥的注射性質(A) 骨碎補 (B) 續斷 28
Fig. 7 (a) 骨碎補濃度之標準曲線 (b) 骨碎補藥物之釋放曲線 30
Fig.8 (a) 續斷濃度之標準曲線 (b) 續斷藥物之釋放曲線 31
Fig. 9 藥物濃度對於細胞生長的影響 (A)骨碎補及續斷 (B) gentamicin 33
Fig. 10 明膠微粒包含藥物對於細胞生長的影響 34
Fig. 11 細胞培養於材料表面上的生長情形 35
Fig. 12 細胞培養於材料表面之鈣化程度 36
Fig. 13 細胞培養於(A) s55 (B) GM (C) GSB (D) XD表面1天之SEM圖 37
Fig. 14 細胞培養於(A) s55 (B) GM (C) GSB (D) XD表面3天之SEM圖 38
Fig. 15 細胞培養於(A) s55 (B) GM (C) GSB (D) XD表面7天之SEM圖 39
Fig. 16 細胞培養於骨水泥表面1及3天的基因表現（A）不同組別的基因表現圖，（B）TNF-alpha、（C）iNOS、（D）IL-1及（E）IL-10的基因表現量 42
Fig. 17 細胞培養於骨水泥表面3及7天的基因表現（A）不同組別的基因表現圖，（B）collagen、（C）ALP、（D）OC及（E）BSP的基因表現量 45

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