二氧化矽奈米纖維基材對於神經幹細胞貼附及分化之影響

The attachment and differentiation of neural stem cell on silica nano fiber

范姜柏人(Po-Zen Fan-Jen)

指導教授 ： 金亭佑

中原大學/工學院/生物科技研究所/碩士(2012年)

https://doi.org/10.6840/cycu201201072

摘要

二氧化矽做為材料利用電紡絲技術將其製成生物三維支架大多主要應用在骨頭組織的修復、再生，而在神經前驅細胞上的影響尚未得到研究，而我們主要利用初級培養的老鼠神經前驅細胞作為研究的模組，根據過去研究室以及文獻研究指出dbcAMP對於神經幹細胞具有誘導分化的特性，因此我們結合兩者去觀察，二氧化矽奈米纖維三維支架是否具有輔助神經幹細胞貼附、分化之功能，之後再利用改質劑使支架帶有胺基觀察輔助之效應。在初步研究中發現神經幹細胞可以貼附在二氧化矽奈米纖維上，並且隨培養的天數觀察細胞數的增生相較聚離胺酸沒有顯著差異，之後處理dbcAMP後發現，改質劑組別細胞在量化神經突長的表現明顯得比聚離胺酸組別來的長，掃描式電子顯微鏡的結果也可發現神經前驅細胞在支架上貼附並分泌胞外基質輔助其生長，而可觀察到神經突長並沒有沿著支架纖維生長而主要是藉由支架的空間去做伸展；之後我利用qRT-PCR偵測在處理dbcAMP五天後的分化表現，發現改質胺基後的二氧化矽奈米纖維有更好的輔助分化效應，綜合以上結果指出二氧化矽本身對於輔助分化沒有特殊效應，但經過改質劑處理後則有明顯的輔助分化效果。

關鍵字

二氧化系奈米纖維 ； 神經球 ； 神經幹細胞

並列摘要

Electrospinning technique using organic and inorganic materials into three dimensional fibrous structures was proven to be useful.
In the past, pointed out that the electrospinning technique using organic and inorganic materials into three dimensional spinning fiber structures is conducive to cell attachment and growth. Silica is a low toxicity, stability of inorganic compounds. Our research aims to find suitable adhesion and differentiation of neural stem cells (NSCs) in silica nanofiber materials. NSCs isolated from 13.5 days of embryonic brains were propagated as neurospheres and culture under organic and inorganic nanomaterial conditions. Silica or Silica conjugated amino groups (Silica-AP) were used as substrates to culture NSCs to examine the effects of these biomaterials on the properties of neural stem/progenitor cells (NSPCs) in the presence of dibutyryl cyclic AMP (dbcAMP) in growth factors-free medium. To test whether the NSCs-derived neurons could express neuronal specific properties, the differentiated cells are characterized by morphological observation, immunostaining and QRT-PCR for neuronal specific proteins. We use the microscopic observation of neural stem cells in different types of nano-materials growth situation. Then we used QRT-PCR identification the mRNA expression level of neuron-specific enolase (NSE) which is neuron labeled protein. Using SEM to identified the morphalogy of neural stem cell. Present results indicate that the silica conjugated amino groups help the differentiation of neural stem cells.

並列關鍵字

neurosphere ； silica nanofiber ； neuralstem cell

參考文獻

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