隨機及順向聚鄰甲氧基苯胺/聚己內酯靜電紡絲法纖維在組織工程之應用探討

The Study of the Application of Random and Aligned Poly(o-methoxyaniline)/Poly(ε-caprolactone) Electrospun Fibers in Tissue Engineering

呂翊帆(Yi-Fan,Lu)

指導教授 ： 李文婷

中原大學/工學院/生物醫學工程研究所/碩士(2016年)

https://doi.org/10.6840/cycu201600146

摘要

本研究利用靜電紡絲技術分別將聚鄰甲氧基苯胺 (Poly(o-methoxyaniline), POMA) 和聚己內酯 (Poly(ε-caprolactone), PCL) 製備成隨機及順向排列之複合式奈米纖維，並摻雜樟腦磺酸 (Camphorsulfonic acid, CPSA)，探討其與心肌母細胞 (H9c2) 的相容性。掃描式電子顯微鏡 (Scanning electron microscopy, SEM) 觀察得知 POMA 纖維直徑為 219±53 nm，添加 PCL 可使其直徑增加，順向電紡與 CPSA 摻雜則會使其直徑下降。接觸角分析發現 POMA 和 PCL 電紡絲具有疏水性表面，而 CPSA 摻雜則會使其表面親水性增加。循環伏安法分析發現未摻雜 CPSA 之電紡絲不具有電活性，摻雜後可使電紡絲導電率提升到半導體範圍。原子力顯微鏡分析發現摻雜 CPSA 電紡絲的楊氏係數較高，而順向電紡絲的楊氏係數又較隨機電紡絲高。拉伸試驗分析發現順向電紡絲的機械性質較隨機電紡絲具有韌性、拉伸強度增加；CPSA 摻雜則會使其剛性增加。將電紡絲浸泡於培養基中 21 天，纖維膨潤但仍可維持纖維型態。將 H9c2 細胞培養於電紡絲上以 SEM 觀察發現，順向纖維可誘導細胞生長方向，而電場方向也會影響細胞本體延展方向。Rhodamine-phalloidin 螢光染色影像可觀察到細胞骨架排列與順向纖維的方向一致。但在隨機電紡絲上，未電刺激組的細胞骨架呈不規則排列；電刺激後，細胞骨架的排列則與電場方向平行。細胞活性分析發現 CPSA 摻雜與 POMA 比例較高的電紡絲中，細胞增生較多；而順向與隨機排列電紡絲的細胞活性則無顯著差異。施予體外電刺激則會抑制細胞生長速度，推測此時細胞可能朝向肌小管分化。本研究證實 POMA 混合 PCL 摻雜 CPSA 可紡出具導電性之順向電紡絲，細胞相容性結果顯示其具有應用於組織工程的潛能。

關鍵字

聚鄰甲氧基苯胺 ； 聚己內酯 ； 靜電紡絲 ； 導電高分子 ； 組織工程 ； 有序奈米纖維

並列摘要

The specific aims of this study were to prepare random and aligned composite nanofibers containing camphorsulfonic acid (CPSA) doped poly(o-methoxyaniline) (POMA) and poly(ε-caprolactone) (PCL) by electrospinning technique as well as explore biocompatibility of electrospun nanofibers with cardiac myoblast (H9c2). The diameter of POMA nanofiber was 219±53 nm as shown by scanning electron microscopy (SEM). Nanofiber diameter increased with increasing PCL content but decreased with CPSA doping and aligned electrospinning process. Contact angle analysis found that surfaces of POMA and PCL nanofibers were hydrophobic and became hydrophilic when doping with CPSA. Cyclic voltammetry showed no electroactivity in electrospun nanofibers without CPSA doping. Conductivity of nanofibers was raised to the range of semiconductor after CPSA doping. Atomic force microscopy analysis revealed that Young’s module of nanofiber was enhanced by CPSA doping and further increased in aligned nanofibers. Tensile test found that aligned electrospun fibers had higher toughness and tensile strength than random fibers, whereas rigidity increased by CPSA doping. The fibers became swollen but still maintained their morphology after soaking in the culture medium for 21 days. SEM images showed that direction of H9c2 cell growth on the aligned scaffolds was parallel to the direction of fiber alignment, whereas it was random on the randomly-oriented scaffolds. Direction of cell growth was also parallel to the direction of electrical field applied. Fluorescent stain of rhodamine-phalloidin found that the arrangement of cytoskeleton was parallel to the direction of fiber alignment, whereas it was random on the randomly-oriented scaffolds. Direction of cytoskeleton was also found to be parallel to the direction of electrical field applied. Cell viability analysis indicated that CPSA doping and high POMA ratio led to higher cell proliferation. No significant difference in cell viability was found between aligned and random nanofibers. In vitro electrical stimulation inhibited cell growth, which might be resulted from myotube differentiation. The study demonstrated aligned and conductive nanofibers of POMA and PCL were successfully prepared. The result of cell compatibility suggest the application potential of conductive nanofibers in tissue engineering.

並列關鍵字

poly(o-methoxyaniline) ； poly(ε-caprolactone) ； electrospining ； conductive polymer ； tissue engineering ； aligned nanofiber

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