利用聚乙烯乙烯醇薄膜接枝雙胺探討其對神經元細胞之影響

本實驗是以乾式法將高分子乙烯乙烯醇(EVAL)聚合物製成薄膜之後，用化學改質的方式將表面氫氧基(-OH)接枝甲基丙烯醯氯(MAC)以間位氯化過氧苯酸(mCPBA)將烯烴雙鍵上進行環氧化反應化後接枝離胺酸(L-lysine)單體和各式不同碳鏈的雙胺。並且將接枝離胺酸(L-lysine)後的薄膜用碳化二亞胺(EDC)修飾離胺酸上的羧酸基接上丁二胺，並利用FT-IR以及接觸角變化分析薄膜改質前後差異和Orange II salt定量膜材表面胺基(-NH2)之接枝密度。同時利用體外細胞培養的方法，來探討材料與細胞之間的生物適合性，將改質前以及改質後的薄膜培養小鼠初代顆粒型神經細胞，以MTT測試來比較細胞的活性並且用光學顯微鏡和掃描式電子顯微鏡來觀察細胞貼附在接枝不同碳鏈的二胺、離胺酸以及經丁二胺修飾之離胺酸並和塗佈聚離氨酸(poly-D-lysine)的乙烯乙烯醇薄膜做比較。小鼠顆粒型神經細胞培養於接枝在經丁二胺修飾過後的lysine其細胞活性表現優於EVAL塗佈聚離胺酸薄膜更勝於未改質薄膜，效果比接枝雙胺類更能提升神經細胞粒線體活性。

關鍵字

聚乙烯乙烯醇；小腦顆粒神經細胞；甲基丙烯醯氯；間位氯；化過氧苯酸；碳化二亞；聚離胺酸

並列摘要

We prepare poly(ethylene-co-vinyl alcohol)(EVAL) membranes by the dry processing ,and then grafted methacryloyl chloride (MAC)on the hydroxyl group of these membrane surface by the way of chemical modification.Furthermore, we grafted different diamines, lysine and modified lysine with 1,4-diaminobutane by EDC/NHS method on them. We used FT-IR and contact angle to assay the material properties after modification then quantification the surface grafted density by orange II salt. We cultured the cerebellar granule neurons of the Wistar rat on the modified membranes and compared with the relative activity by MTT assays. In addition, the cell morphology was observed by optical microscope and scanning electron microscope (SEM). The modification of the EVAL with lysine modified 1, 4-diamino -butane, in general, have better relative activity than coating PDL. We observe this method could effectively induce the growth of neuron cells.

並列關鍵字

cerebellar granule cells ； methacryloyl Chloride ； EDC ； PDL ； 1,4-diaminobutane ； EVAL ； lysine

參考文獻

1. K.E. Crompton , J.D. Goud and R.V. Bellamkonda , “Polylysine-functionalished thermoresponsive chitosan hydrogel for neural tissue engineering”, Biomaterial , 28 , 441-449 , 2007

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國際替代計量

利用聚乙烯乙烯醇薄膜接枝雙胺探討其對神經元細胞之影響

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