幾丁聚醣/聚外消旋乳酸電紡絲複合纖維支架結構對人類纖維肉瘤細胞貼附與增生之研究

Study on the structure of electrospun chitosan/ poly(DL-lactide) composite fibrous scaffold for human fibrosarcoma cells (HT1080) attachment and proliferation

陳聖翰(Sheng-Han Chen)

指導教授 ： ARRAY(0xac68e50)

中原大學/工學院/化學工程研究所/碩士(2010年)

https://doi.org/10.6840/cycu201000941

摘要

本研究主要探討幾丁聚醣(chitosan)/聚外消旋乳酸(PDLLA)電紡絲複合纖維支架結構設計對人類纖維肉瘤細胞(human fibrosarcoma cells，HT1080)貼附(attachment)與增生(proliferation)行為之影響。研究中首先藉由控制(1)紡絲參數的變化，在PDLLA高分子濃度：15 wt%、工作距離：30 cm與針頭直徑：0.16 mm的條件下，可紡製出bead-free之PDLLA纖維結構型態，且同時提升整體纖維直徑的均一性(uniformity)。其次，對於(2)紡絲溶液性質之探討，經由chitosan紡絲溶液進行降解測試發現，當降解時間達10天時，藉由黏度的下降和chitosan-PEO分子鏈糾結程度之增加可大幅地改善chitosan的紡絲性與纖維結構均勻性，而在導入旋轉滾筒轉速為1000 rpm的條件下，所收集之chitosan纖維具有較佳的順向性。
由MTT assay細胞存活率測試結果顯示chitosan/PDLLA複合纖維的立體結構相較於PDLLA平板薄膜和cover slip的平面結構而言，可使人類纖維肉瘤細胞具有較高的增生能力，且經由雷射共軛焦顯微鏡(LSCM)觀察發現細胞除了可貼附並生長於random PDLLA纖維所建構的立體空間中，更由confocal立體影像證實細胞有滲入(infiltration)至膜內生長的趨勢，此外，當順向性chitosan纖維覆蓋率增加至12.66 g/cm2時，可使貼附的細胞進行沿著順向性chitosan纖維軸向進行生長，提供引導細胞生長方向的功能，由以上分析結果指出利用chitosan/PDLLA複合纖維支架結構之設計，可成功促進人工支架與細胞之間的生物相容性(biocompability)。

關鍵字

幾丁聚醣 ； 聚外消旋乳酸 ； 細胞外基質 ； 複合纖維支架 ； 電紡絲 ； 組織工程

並列摘要

In this study, the effect of morphology design of chitosan/ poly(DL-lactide) electrospun composite fibrous scaffold on the attachment and proliferation of human fibrosarcoma cells (HT1080)were investigated. The bead-free and the most uniformity of PDLLA fibers could be fabricate through adjusting spinning parameters, e.g. under the condition of polymer concentration: 15wt%, working distance: 30 cm and needle diameter: 0.16 mm. Otherwise, the degradation test of chitosan spunning solution were also investigated. The result showed that the viscosity of chitosan solution decrease with increasing the degradation time. After 10 days of degradation, the spinnability and morphology of chitosan fibers were dramatically improved. The better alignment of chitosan fibers were collected successfully at 1000 rpm of rotating-drum collector.
MTT assay results indicated the 3D structure of chitosan/ PDLLA composite fibrous scaffold could promote the ability of human fibrosarcoma cells proliferation compare with the 2D structure of PDLLA film and cover slip. When the collecting amount of chitosan aligned fibers increased to 12.66 g/cm2, the growth of cells were guided and elongated by the aligned chitosan fibers. And the 3D conforcal images showed that the cell growth could infiltration to the inside of fibrous scaffold, it’s consistent with the structure design of chitosan/PDLLA composite fibrous scaffold. According to the principle of tissue engineering, chitosan/PDLLA composite fibrous scaffold could have excellent potential for tissue regeneration and wound healing applications in the future.

並列關鍵字

composite fibrous scaffold、poly(DL-lactide) ； chitosan ； extracellular matrix (ECM) ； tissue engineering ； electrospinning

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