不同材料神經導管中周邊神經再生情形及相關蛋白質表現評估;Evaluation of Regenerated Nerves and Their Protein Expression in Three Different Conduit Materials

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Title:	不同材料神經導管中周邊神經再生情形及相關蛋白質表現評估;Evaluation of Regenerated Nerves and Their Protein Expression in Three Different Conduit Materials
Authors:	陳力殊;Li-Shu Chen
Contributors:	中國醫藥大學：中國醫學研究所碩士班
Keywords:	周邊神經再生;降解性神經導管;明膠;綠梔子素;西方墨點法;Peripheral nerve regeneration;Biodegradable neural guide conduits;Gelatin;Genipin;Western-blot
Date:	2009-07-21
Issue Date:	2009-08-10 16:01:09 (UTC+8)
Abstract:	可降解性之生醫組織材料具有不需二次手術取出以免傷害神經之優點，故用於製作神經導管僑接周邊神經可使臨床後遺症減少。在眾多可降解性之生醫組織材料中，明膠為膠原蛋白之二級結構破壞後的產物，具便宜、抗原性低及毒副作用低等優點。本實驗以從中藥梔子萃取之梔子素(ginipin)及EDC/NHS〔1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/ N-hydroxysuccinimide〕二種不同交聯劑交聯而得的明膠管作為可降解性神經再生導管的材料（分別稱之為GGC導管及ENG導管），並以非降解性之矽膠管做對照，將三種導管殖入大鼠被截斷之坐骨神經之間，手術八週後，以組織切片評估三種神經導管對神經生長功能的影響；並同時取再生之神經組織，經研磨分離出蛋白質後，以西方墨點法分析在神經再生過程中會大量表現之蛋白質synapsin Ι、GAP-43以及和組織纖維化相關的TGF-β1之表現量，嘗試以分子角度評估導管材料的好壞。組織切片結果顯示，矽膠管組大鼠的再生神經成熟度最佳，ENG組次之，GGC組最差。蛋白質分析結果，三組之synapsin及GAP-43表現量皆以矽膠管組最多，ENG組次之，GGC組最少，和組織切片之結果相符合；TGF-β1之表現量則呈現ENG組最多，GGC組次之，矽膠管組最少，此結果則無法單就組織切片中纖維疤痕組織之厚度做解釋。 In the present study, we used genipin-crosslinked gelatin conduit (GGC) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide / N-hydroxysuccinimide cross-linked gelatin conduit (ENG) to bridge a 10 mm gap in the sciatic nerve of the Sprague-Dawley rat. These two biodegradable materials are both successful apparatus to reconstruct nerve gaps, and can avoid the disadvantage of a second surgery. However, the regenerated nerve in GGC and ENG were less mature than the regenerated nerve in non-biodegradable silicone tube. Moreover, the scar tissue at the outer area of the regenerated nerve caused by degrading materials could have harmful effect to the regenerating nerve. In this study, we compared the effectiveness of three conduit materials, named silicone, GGC and ENG, in repairing a 10 mm gap in the rat sciatic nerve. Eight weeks after implantation, their effectiveness as a guidance channel were evaluated by morphometric data and protein analysis of the regenerated nerve. As a result, we found that the regenerated nerves in the silicone conduits had the most mature structure, whereas the regenerated nerves in the GGC conduits were the most immature one. In addition, the higher expression of nerve regenerating associate protein synapsin and GAP43 in silicone group also indicated that the regenerating process in silicone conduits were more vigorous. On the other hand, the level of scar formation associated protein TGF-β1 was lower in the silicone group than the other two groups, which suggested the harmful effect of scar tissue in nerve regeneration.
Appears in Collections:	[Graduate Institute of Chinese Medical Science] Theses & dissertations

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