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標題: | 表面接枝兒茶素及釋放控制洛伐他汀之幾丁聚醣再生膜對牙周組織再生及癒合之促進 Chitosan Membrane Grafted with Epigallocatechin-3-Gallate and Controlled Release of Lovastatin for Periodontal Tissue Regeneration |
作者: | Wei-An Shih 施維安 |
指導教授: | 陳漪紋(Yi-Wen Chen) |
關鍵字: | 牙周病,再生膜,幾丁聚醣,EGCG,lovastatin,組織再生, periodontal disease,GTR membrane,chitosan,EGCG,lovastatin,tissue regeneration, |
出版年 : | 2014 |
學位: | 碩士 |
摘要: | 可吸收性再生膜因具有生物相容性及促進組織修復能力,已被廣泛運用於牙科再生手術。本研究利用生物可降解性材料-幾丁聚醣(Chitosan)作為再生膜基材,製備出緻密的三層膜結構,表層接枝兒茶素(EGCG),而中間層則作為洛伐他汀(Lovastatin)的釋放載體。進行微拉伸測試、FTIR-ATR分析、鹼性磷酸酶(ALPase)活性測試、細胞毒性、貼附及增生等試驗。結果顯示再生膜厚度為0.17 mm,強度為1.67MPa,為目前臨床使用膠原蛋白再生膜的2倍。以丁二酸酐(succinic anhydride)修飾EGCG後透過EDC/NHS交聯於幾丁聚醣再生膜之表面,FTIR檢測出在1215 cm-1處的芳香氫氧基吸收峰,顯示EGCG以化學鍵結方式接枝於幾丁聚醣再生膜表面。細胞測試結果顯示,以14mM EGCG濃度進行接枝反應,能增加幾丁聚醣再生膜的細胞親和性,促進人類牙齦纖維母細胞貼附及增生。而再生膜中間層搭載1mg/cm2 lovastatin,其以滲透方式釋放至磷酸緩衝液中,能達到21天的長時間釋放,與人類胚胎上顎間質細胞共同培養後會先抑制其增生,並且在第7天時提升骨分化酵素ALPase的活性,促進細胞進行骨分化。表面接枝EGCG並搭載lovastatin的幾丁聚醣再生膜能促進傷口癒合及骨質新生,對於牙周組織的修復及再生極具潛力。 Biodegradable membrane for guided tissue regeneration (GTR) has been utilized as a barrier in periodontal surgery to promote tissue healing. As epigallocatechin-3-gallate (EGCG) can reduce inflammation and lovastatin can increase bone regeneration, the aim of this study was to prepare a functional tri-layer chitosan membrane to enhance the healing of periodontal tissue. EGCG was grafted on the outer surface of the top layer and lovastatin was embedded in the middle layer for controlled release. EGCG was grafted on the outer surface using ethyl (dimethylaminopropyl) carbodiimide/ N-hydroxysuccinimide (EDC/NHS) reaction. The microtensile strength of chitosan membrane was measured and Fourier transform infrared spectroscopy (FTIR) was used for surface characterization. UV–vis spectroscopy was used to quantify lovastatin release. Cell adhesion, proliferation, nitric oxide, and alkaline phosphatase (ALPase) assay were evaluated for biological function. The microtensile strength of fabricated membrane was 1.67 MPa which was superior to commercial collagen GTR membrane. FTIR spectra indicated EGCG was grafted on membrane as the 1215 cm-1 peak corresponding to aromatic C-O was identified. The chitosan membrane grafted by 14mM EGCG could increase the adhesion and proliferation of human gingival fibroblasts to promote the tissue wound healing. The membrane which loaded with 1mg/cm2 lovastatin could release for 21 days, and inhibited the cell proliferation at 5th day followed by increasing the ALPase activity at 7th day in Homo sapiens palatal mesenchymal cells. The chitosan membrane can promote bone activity and wound healing. It has potential to be used in GTR surgery for enhancing periodontal repair. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57177 |
全文授權: | 有償授權 |
顯示於系所單位: | 口腔生物科學研究所 |
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