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    請使用永久網址來引用或連結此文件: http://ir.lib.ncu.edu.tw/handle/987654321/6363


    題名: Galectin-1蛋白促進老鼠軟骨細胞於幾丁聚醣修飾之聚乳酸-聚乙酸醇共聚物支架生長之研究;Galectin-1 Stimulates Immortal Rat Chondrocyte Proliferation on the Surface of Chitosan-coated PLGA Scaffold
    作者: 段維杰;Wei-Cheh Tuan
    貢獻者: 生命科學研究所
    關鍵詞: 幾丁聚醣;聚乳酸-聚乙酸醇;軟骨細胞;組織工程;chitosan;PLGA;chondrocyte;galectin-1;tissue engineering
    日期: 2004-07-07
    上傳時間: 2009-09-22 10:18:15 (UTC+8)
    出版者: 國立中央大學圖書館
    摘要: 組織工程研究組織與器官之再生與形成,利用材料科學與生物科技,在一個模仿組織與器官形狀的材料中植入細胞,使細胞依著模型來長成新的組織與器官,以供修復人體缺損。目前廣泛使用之生醫材料聚乳酸-聚乙酸醇【polylactide- co-glycolide (PLGA)】可藉由新的機械製程-熱融熔成型法製備成具有精確孔洞大小、分佈均勻之立體支架。本研究利用幾丁聚醣披覆於PLGA支架上(chitosan-PLGA),再利用Galectin-1(GAL1)與PLGA支架上之幾丁聚醣結合,作為軟骨細胞生長之支架(GAL1-chitosan-PLGA)。實驗結果發現幾丁聚醣披覆可有效增加PLGA支架之膨潤度及吸水性,顯示幾丁聚醣披覆可降低PLGA支架表面疏水性質;結果也顯示GAL1蛋白能自動吸附於chitosan-PLGA立體支架上,並促進老鼠軟骨細胞IRC生長於chitosan-PLGA支架,於GAL1- chitosan-PLGA支架上之細胞外觀較偏圓形且易聚集生長,而GAL1蛋白促使細胞貼附後一小時造成ERK磷酸化能力降低。GAL1蛋白促進細胞於chitosan-PLGA支架增生能力會因細胞酪胺酸磷酸化大量表現而加速,然而當絲胺酸/息寧胺酸的磷酸化被抑制,細胞存活和促進增生作用會被明顯抑制。本實驗證實幾丁聚醣及GAL1蛋白結合PLGA之立體支架可有效促使軟骨細胞之貼附及生長,可應用作為軟骨組織工程的生物因子。 Tissue engineering employs the principles and methods of engineering and life sciences toward fundamental understanding of structure function relationship in normal and pathological mammalian tissues as well as the development of biological substitutes for the repair or regeneration of tissue or organ function. The widespread biomaterial polylactide-co-glycolide (PLGA) could fabricate by new condition mechanical system of rapid prototyping system prepared the 3D biocompatible scaffold with the advantage of precise pore size and distribute higher characteristic. In this study, we investigated the application of chitosan- and GAL1-modified PLGA in tissue engineering. The results show that the appearance of chitosan modified PLGA scaffold was extensively expansive than PLGA after immersing on ddH2O for 2 weeks and water absorption ratio was also 70% higher than PLGA . This result indicated that the chitosan-modified PLGA would be more hydrophilic than PLGA itself. Experimental results also showed that GALl can be efficiently and spontaneously coat on the surface of chitosan-PLGA scaffold to promote the adhesion and proliferation of IRC cells. IRC adhesion and proliferation on GAL1-chitosn- PLGA scaffold was dose-dependently inhibited by the presence of TDG (the specific inhibitor of galectin on CRD). These results strongly suggested that GAL1 plays a critical role in enhancing IRC cells adhesion on chitosan-PLGA scaffold. IRC growing on GAL-chitosan-PLGA tend to form cell-cell aggregation on the surface, an indicating of tissue differentiation. In addition, level of ERK phosphorylation were inhibited 1 hr after IRC cultured on GAL1-chitosan-PLGA scaffold. The adhesion and proliferation of IRC on GAL1-chitosan-PLGA scaffold were enhanced by tyrosine phosphorylation, but attenuated by serine/threonine phosphorylation. In conclusion, the present results suggest that both chitosan and GAL1 play important role in enhancing IRC cell adhesion and proliferation on PLGA scaffold, and GAL1-potentiated cell adhesion and/or proliferation might associate with cellular phosphorylation.
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