【摘要】：神經(jīng)損傷后可植入性組/細胞支架設計是周圍神經(jīng)損傷、和脊髓損傷后神經(jīng)修復一個重要思路。本研究利用靜電紡絲技術成功構建具有隨或有序拓撲結構聚甲丙烯酸甲酯（PMMA）納米纖維材料，并進行大鼠原代背根神經(jīng)節(jié)神經(jīng)元（DRGn）培養(yǎng)，及其與大鼠原代雪旺細胞（SCs）共培養(yǎng)。結果顯示所構建PMMA電紡納米纖維具有良好生物相容性，適合進行神經(jīng)細胞培養(yǎng)；通過所轉染綠色熒光蛋白表達，發(fā)現(xiàn)DRGn能夠順利在電紡纖維上貼壁并生長，電紡纖維拓撲結構能夠影響DRGn生長能力和生長方向，DRGn在有序電紡納米纖維上能夠生成更長神經(jīng)突，且神經(jīng)突生長方向能夠和及質(zhì)纖維延伸方向保持高度一致；通過進一步分析及掃描電鏡檢查，發(fā)現(xiàn)不管在隨或有序PMMA電紡納米纖維，DRGn神經(jīng)突均能夠沿纖維行，，但相比隨纖維，有序電紡纖維對DRGn引導能力更強，DRGn似乎更傾向于循有序纖維生長；通過DRGn與SCs共培養(yǎng)，則發(fā)現(xiàn)較PMMA薄膜和隨電紡纖維，有序電紡纖維能夠在形態(tài)上促成DRGn與SCs建立更多共定位，估計能夠促進后續(xù)髓鞘化進程。本課研究結果示有序PMMA電紡納米纖維具有作為神經(jīng)損傷后植入性支架力，值后續(xù)更進一步體內(nèi)研究。
[Abstract]:The design of implantable group / cell scaffold after nerve injury is an important idea for peripheral nerve injury and nerve repair after spinal cord injury. In this study, poly (methyl methacrylate) (PMMA) nanofibers with random or ordered topological structure were successfully constructed by electrospinning technique, and the primary dorsal root ganglion neurons (DRG) of rats were cultured with (DRGn). And co-cultured with rat primary Schwann cells (SCs). The results showed that the PMMA electrospun nanofibers had good biocompatibility and were suitable for nerve cell culture. Through the expression of green fluorescent protein, it was found that DRGn could attach and grow on electrospun fibers smoothly. The topological structure of electrospun fibers could affect the growth ability and growth direction of DRGn, and DRGn could produce longer nerve processes on ordered electrospun nanofibers. And the direction of neurite growth was highly consistent with the direction of fiber extension. By means of further analysis and scanning electron microscopy, it was found that the DRGn nerve processes were able to walk along the fibers, regardless of whether they were electrospun with or without PMMA, but the ordered electrospun fibers had stronger ability to guide DRGn than with fibers. DRGn seems to be more inclined to follow the order of fiber growth; By co-culture of DRGn and SCs, it was found that the ordered electrospun fibers could promote more co-localization of DRGn and SCs in morphology than those of PMMA thin films and electrospun fibers, and could promote the subsequent myelinization process. The results of this study show that ordered PMMA electrospun nanofibers have implantable scaffold force after nerve injury and further study in vivo.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R651.3;R318.08

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