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    National Tsing Hua University Institutional Repository > 生命科學院  > 分子醫學研究所 > 博碩士論文 >  利用表面電漿共振儀研究固定式表皮生長因子於骨組織工程之應用


    Please use this identifier to cite or link to this item: http://nthur.lib.nthu.edu.tw/dspace/handle/987654321/85691


    Title: 利用表面電漿共振儀研究固定式表皮生長因子於骨組織工程之應用
    Authors: 袁知敏
    Yuan, Chih-Min
    Description: GH02101080604
    碩士
    分子醫學研究所
    Date: 2014
    Keywords: 表面電漿共振儀;表皮生長因子;固定化
    Surface Plasmon Resonance;Epidermal Growth Factor;Immobilizatiom
    Abstract: 固定表皮生長因子已被證實具有抑制胞吞現象即調控細胞增生與分化的功能。在本研究中將依序分析固定後的表皮生長因子於蛋白質-蛋白質間結合的狀態、細胞訊息啟動、組織修復的影響。因此我們探討三個與固定相關的因素:(1)固定密度的影響。(2)固定方式的影響。(3)添加中藥小分子牡丹皮增強結合能力的影響。
    在蛋白質-蛋白質間結合狀態結果顯示,當晶片上固定密度在50 pg/mm2-1000 pg/mm2其親和力是不受到密度影響,並且固定量越多所能結合的受體也越多。具有適當的方向固定方式可以維持表皮生長因子的活性60%長達16天以及啟動受體二聚化的作用。在細胞試驗中,發現磷酸化的現象成功被啟動以及動物實驗中新生骨組織的增加。最後利用添加牡丹皮發現能夠增強10倍的親和力及具有更好的骨組織修復程度。因此我們推論,適當調控表皮生長因子與受體結合的機制對於之後的訊息傳遞現象是息息相關且非常重要的。
    Immobilization of Epidermal Growth Factor (EGF) has been shown to inhibit endocytosis and regulate cell proliferation and cell differentiation. However, nonspecific immobilization of EGF on substrate has been demonstrated to drastically diminish EGF bioactivity. In this thesis, we would like to study immobilized EGF property and binding ability by protein-protein interaction, cell signaling transduction and tissue formation. Three factors of EGF immobilization will be discussed: (1) Density of immobilization. (2) Linkage of immobilization. (3) EGF-EGFr binding enhancement by Moutan Cortex.
    The protein-protein interaction result showed that immobilized EGF density with 50-1000 pg/mm2 did not affect its affinity and activity. Immobilized EGF with appropriate orientation on SPR chip can maintain its function and activity for 16 days, inducing EGFr phosphorylation in cell experiment and more new bone tissue formation in bone repair assessment. In the case of binding enhancer treatment, Moutan Cortex enhanced EGF and EGFr 10-fold binding affinity in SPR experiment and better bone repair in animal experiment. These results supported that affinity enhancement can induce more cell signaling transduction for tissue regeneration. Therefore, the regulation of EGF-EGFr interaction is significant for signaling transduction and bone formation.
    URI: http://nthur.lib.nthu.edu.tw/dspace/handle/987654321/85691
    Source: http://thesis.nthu.edu.tw/cgi-bin/gs/hugsweb.cgi?o=dnthucdr&i=sGH02101080604.id
    Appears in Collections:[分子醫學研究所] 博碩士論文

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