靜電紡絲製備具生物相容性之二氧化矽奈米纖維及其於神經組織工程之應用

Preparation of the biocompatible silica nanofibers by an electrospinning method for application of nerve tissue engineerings

簡郁珊(Yu-Shan Chien)

指導教授 ： 陳玉惠

中原大學/理學院/化學研究所/碩士(2014年)

https://doi.org/10.6840/cycu201400799

摘要

本研究係利用溶膠-凝膠法以靜電紡絲技術製備二氧化矽奈米纖維，並修飾成具有生物活性之生物性支架。製備過程以二氧化矽前驅溶液與Polyvinylpyrrolidone (PVP增稠劑) 均勻混合，為紡絲用前驅溶液，並於高電壓環境下進行紡絲，而得二氧化矽-PVP奈米纖維 (SNFP)，經熱處理移除PVP增稠劑後，可得立體絲狀結構且具多孔性之二氧化矽奈米纖維 (silica nanofiber, SNF)。接著，二氧化矽奈米纖維以胺基官能化後，使其為生物活性之多孔立體生物支架 (SNF-AP3)。接著以Sulfosuccinimidyl – 4 – (N-maleimidomethyl) cyclohexane – 1 – carboxylate (Sulfo-SMCC) 作為交聯劑，分別以化學接枝及物理吸附方式修飾層黏連蛋白 (laminin, L) 固定於SNF-AP3材料上，來探討修飾後之材料對神經幹細胞生長及分化的影響。其次，以純二氧化矽前驅溶液，進行靜電紡絲技術製備奈米纖維，成功得到纖維型態，即純二氧化矽奈米纖維 (pure silica nanofiber, PSNF)，並與SNF系列比較材料降解型態。
研究過程中除利用SEM觀察其表面形貌及FTIR、TGA、BET、接觸角及降解行為，探討其物理、化學性質外，並利用免疫螢光染色技術以螢光顯微鏡的方法對於層黏連蛋白在纖維上的分布情形進行觀察。結果顯示SNF-AP3/L及SNF-AP3/SL進行神經幹細胞培養，能有效分化成神經元，因此這兩種材料有做為生物支架之潛力。降解性之比較，證實SNF系列屬於生物可降解性之材料， PSNF系列則在短期間無明顯之降解性，未來可參雜無毒性之模板，製備成具孔洞之結構調整其降解能力，並加入功能性載體，以期做為具有潛力之生物醫學材料。

關鍵字

靜電紡絲 ； 神經幹細胞 ； 二氧化矽 ； 層黏連蛋白 ； 降解

並列摘要

In this study, the silica nanofiber was prepared via the sol-gel polymerization by the electrospinning method. In the process, polyvinylpyrrolidone (PVP) was mixed with the silica precursor as a thickener to form the electrospinnable solution for the electrospinning process. The polymer was then removed by the calcination to obtain the porous silica nanofiber (SNF). Subsequently, the as-prepared SNF was modified with (3-aminopropyl) trimethoxysilane (APTS) to form the amino functionalized SNF (SNF–AP3) for application in the neural tissue engineering.
SNF–AP3 was further modified by sulfosuccinimidyl–4–(N-¬maleimido-methyl)¬¬cyclohexane–1–carboxylate (Sulfo-SMCC) as a crosslinking agent, followed by grafting Laminin, a growth factor for neuron stem cell, on the SNF-AP3 fiber. The as-prepared chemically grafting Laminin SNF (SNF-AP3/SL) was then compared to the physical adsorpted SNF-AP3 fiber (SNF-AP3/L) for the impact on the growth and differentiation of neural stem cell. In addition, pure silica nanofibers (PSNFs) were also successfully prepared by the electrospinning method with the silica precursor solution without polymer thickener.
All the structures and properties of the as-prepared SNF, the modified SNF (SNF–AP3、SNF-AP3/L and SNF-AP3/SL) and PSNFs were confirmed by a combination of FTIR, TGA, nitrogen adsorption/desorption isotherms, contact angle analysis and SEM measurements. The distributions of the laminin on the surface of SNF-AP3/L and SNF-AP3/SL fibers were investigated with the immunofluorescence technique. The results showed that the neural stem cells could effectively differentiate into neurons on both SNF-AP3/L and SNF-AP3/SL. Therefore, these two materials have the potential as biological scaffolds. Besides, the degradation behaviors of the as-prepared pure silica nanofibers (PSNF) were compared with that of the SNF series materials. The results indicated that the SNF series fibers were biodegradable, while PSNF series is relatively insoluble due to non-porous structure. In the future, a porous PSNF may be obtained by using non-toxic and water soluble materials as templates, leading to be potential of biodegradable materials.

並列關鍵字

silica ； laminin ； biodegradable ； neural stem cell ； electrospinning

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