微細纖維狀仿生薄膜之製作及其作為眼組織用基質 之探討

Fabrication of biomimic fine-fibrous membrane as a matrix for eye tissue engineering

簡安儀(An-I Chien)

指導教授 ： 曾厚

臺北醫學大學/醫學院/醫學科學研究所/碩士(2006年)

摘要

近年來，利用羊膜為基底，培養角膜上皮細胞、輪部幹原細胞，作為角膜之傷口修復與重建，是眼科手術上的一大突破；另一方面，電氣紡絲技術因製程單純、製備容易也在近年組織工程領域中被廣泛研究與應用，而利用電氣紡絲製備出的纖維狀薄膜，其纖維型態又與去除上皮細胞層的羊膜基底層極為相似；因此本研究欲結合此兩種概念應用於眼組織工程之上。實驗是以生物可吸收性高分子PLLA、PBSA及PEO為原料，利用電氣紡絲技術，製備仿羊膜之纖維薄膜，製程中控制不同變因(射源、輸液速度、被覆材料、薄膜處理等)，製備表面性質不同之薄膜，探討角膜細胞之貼附、增生情形，並以細胞生長情形較佳之薄膜進行角膜細胞多層化試驗。薄膜性質以偏光顯微鏡、孔洞測定儀、掃描式熱分析儀與X射線繞射儀分析；之後再擇優將角膜上皮細胞培養於電紡薄膜上，觀察細胞之貼附與增生速度。由實驗結果可知，若將幫浦輸液速度設定為2ml/hr電紡3%PLLA之溶液，所製備之薄膜纖維直徑範圍界於0.42~1.1?m，平均直徑為0.66?m，而利用1N 之氫氧化鈉處理薄膜20分鐘後，薄膜之纖維型態改變、親水性增加，並能增加角膜細胞之貼附與增生數量，但材質變得較為易碎，另外膠原蛋白披覆之方法亦能幫助SIRC的貼附與增生，但其纖維型態與強度則無明顯改變。

關鍵字

電氣紡絲、組織工程、角膜上皮細胞、生物可吸收性高分子

並列摘要

Cultured epithelial cell or epithelial stem cell on amniotic for corneal repair and reconstruction was a great progress on ophthalmology surgical in recent years. On the other hand, electrospinning is an easy and simple polymer processing technology. Its non-woven fiber architecture is almost similar human acellular amniotic membrane. In our study, these two concepts were combined for eye tissue engineering. Bioabsorbable polymers (PLLA, PBSA, PEO) were used as materials to fabricated fine-fibrous membranes via the electrospinning process. We also prepared membranes with different surface features by controlling factors (two-feeding, feeding rate, coating proteins, and NaOH treated). To investigated which one is better for corneal epithelial cell’s attach and proliferating ability. The structure and morphology of electrospun membranes were investigated by scanning electron microscopy (SEM), differential scanning calorimeter (DSC), and X-ray diffraction scattering, contact angle detector. SIRC were cultured on the original and modified membrane and the cell morphology, attachment, proliferation were studied. We demonstrated that PLLA solution (3%) in a mixture solvent of Dichloromethane / Dimethylformamideb (9:1) was electrospun into non-woven fiber mesh with the fiber diameter ranging from 0.42 to 1.1?m. PLLA and PBSA membranes treated in 1N NaOH solution for 20 minutes can improve SIRC’s adhesion and proliferation ability. Stretch strength test revealed significant differences between treated and non-treated membranes than that of non-treated. And NaOH-treated scaffolds exhibited more surface roughness. The collagen coating method also can obviously improve cell adhesion and proliferation, and their mechanical strength is better than NaOH-treated membrane.

並列關鍵字

Electrospinning ； Tissue engineering ； Corneal epithelial cell ； bioabsorbable

參考文獻

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