綠梔子素交聯明膠製造多孔及無孔性神經導管之周邊神經再生與相關蛋白質表現評估;Evaluation of Regenerated Nerves and Their Protein Expression in Porous and Nonporous Genipin-Cross-linked Gelatin Conduits

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Title:	綠梔子素交聯明膠製造多孔及無孔性神經導管之周邊神經再生與相關蛋白質表現評估;Evaluation of Regenerated Nerves and Their Protein Expression in Porous and Nonporous Genipin-Cross-linked Gelatin Conduits
Authors:	詹瞿霙;Chu-Ying Chan
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;biodegradable nerve guide conduits;gelatin;genipin
Date:	2006-07-24
Issue Date:	2009-08-10 16:01:13 (UTC+8)
Abstract:	周邊神經損傷的修復是學者們致力研究的方向，藉由神經導管（nerve conduit）引導斷端神經再生為一理想的修復方式，其中，使用降解性材料製成之神經導管，具有避免導管植入後需再次開刀取出的優點，為近年來周邊神經修復的熱門研究主題。在眾多降解性材料中，明膠（gelatin）是由膠原蛋白經水解加熱處理而來，免疫排斥性較低，具有良好的生物相容性。綠梔子素（genipin）是從中藥梔子果實中萃取出的天然交聯劑，細胞毒性較低，並可增強材料的機械強度，延長材料的使用時間。本研究以明膠為基材、綠梔子素為交聯劑製造多孔性（PGGC）及無孔性（GGC）神經導管，比較PGGC、GGC與矽膠導管三者引導SD大鼠坐骨神經缺損10 mm間距的神經再生情況，並利用西方墨點法分析相關蛋白質GAP43、synapsin I和TGF-β1表現，術後八週再生神經組織通過間距比率矽膠導管組為54％，PGGC、GGC通過間距比率為100％。組織切片結果矽膠管組再生神經總面積最大，PGGC組和GGC組再生神經周圍有一層纖維化結締組織，此結果與兩組TGF-β1表現量較高有相同的趨勢。另外，PGGC組組織切片再生神經成熟度與GAP43表現量均優於GGC組，證明PGGC的通透性較有利於神經再生。而在PGGC和矽膠導管比較方面，PGGC組的GAP43表現量較高，再生神經組織的生長和萌芽情況較佳，但若考量到周邊纖維化結締組織對神經功能恢復與成熟度的影響，以synapsin I表現量較高的矽膠導管組，再生神經組織末端的成熟度較好。 The way to improve peripheral nerve regeneration has been devotedly studied by the researchers. Using the nerve conduits to bridge axonotmesis has been proven as a practical repairing method. The nerve conduits made of biodegradable materials can avoid a second surgery to remove the non-degradable conduits. Therefore, the subjects become popular research area. Among the biodegradable materials, gelatin derived from collagen posses the characteristic of lower antigenicity and better biocompatibility. Genipin extracted from the Chinese herb-Gardenia jasminoides Ellis is a natural substance and low-cytotoxic cross-linking agent. In the presented study, biodegradable genipin-cross-linked gelatin conduits (GGCs) and highly permeable genipin-cross-linked gelatin conduits (PGGCs) were fabricated for next assays. GGC, PGGC and non-degradable silicone tubes were all to be evaluated of the capability of bridging a 10mm gap within the sciatic nerve of the Sprague-Dawley rat. The expression of GAP43, synapsin I and TGF-β1 was also compared in this study. Overall gross examination of the GGCs and the PGGCs after 8 weeks of post implantation revealed 100％ of nerve formation in the tubes, and the silicone tubes only demonstrated 54％, respectively. The histological study further showed that the regenerated nerve area in the silicone tubes was the greatest. Abundant scar tissues surrounding the regenerated nerves were obviously observed in the PGGCs and GGCs. Coordinately, these nerves also showed apparent expression of TGF-β1 than that in the non-degradable silicone tubes. In addition, the regenerated nerves in the PGGCs demonstrated relatively the characteristic of more maturity and had much more expression of GAP43 than those in the GGCs. These data suggested that the PGGCs could offer effective aids for regenerating nerves compared with the GGCs. Compared with the regenerated nerves in the silicone tubes, the nerves in the PGGCs expressed high levels of GAP43 and had better regeneration ability. However, considering the impedance of scar tissue at the nerve repairing site, the silicone tubes expressed high levels of synapsin I, which implied better nerve terminal maturation.
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