聚鄰甲氧基苯胺電紡絲於組織工程之應用

Study of Electrospun Poly (o-methoxyaniline) Nanofibers for Tissue Engineering

謝沐峰(Mu-Fong Hsieh)

指導教授 ： 李文婷

中原大學/工學院/醫學工程研究所/碩士(2009年)

https://doi.org/10.6840/CYCU.2009.00818

摘要

摘要
導電高分子同時具有有機高分子的物理和化學特性以及金屬的導電性，已被應用在光電材料及生物感測上。本研究旨在探討以電紡方式所製備的聚鄰甲氧基苯胺奈米絲薄膜之生物相容性及其在組織工程應用之可行性。掃瞄式電子顯微鏡結果發現聚鄰甲氧基苯胺奈米絲直徑在 200 ~ 300 nm；表面接觸角為 104 ± 8¬ o，為疏水性結構；熱重分析儀分析得知其裂解溫度在300 ~ 600 oC；循環伏安法分析得知本材料具有氧化還原之特性。根據 ISO 10993-5 的規範，我們發現聚鄰甲氧基苯胺奈米絲薄膜的間接細胞毒性，不論是在電刺激前後，皆屬於生物體可接受的範圍。L929 小鼠纖維母細胞、C2C12 小鼠肌原母細胞之貼附與增生結果發現，聚鄰甲氧基苯胺奈米絲薄膜對於C2C12細胞的貼附以及增生均較L929細胞來的佳；進一步探討C2C12 細胞之MyoD、Myogenin、Myf-5、MRF4基因表現，初步證實聚鄰甲氧基苯胺奈米絲薄膜可促進 C2C12 細胞之分化能力。利用巨噬細胞探討材料之體外生物相容性，發現聚鄰甲氧基苯胺奈米絲薄膜會誘發巨噬細胞釋放些微一氧化氮與發炎激素；進ㄧ步量測 LDH 的釋放量證明材料對細胞會造成些微損傷，引起些微發炎反應，但仍屬可接受之範圍。蛋白質吸附試驗發現，此材料在與人類血清白蛋白以及血漿纖維蛋白原反應 48 小時之吸附量達 80 μg/cm2。將聚鄰甲氧基苯胺奈米絲薄膜植入 Wistar 大鼠背部皮下，ㄧ及四週之組織切片中發現有些許嗜中性球、巨噬細胞的聚集，八及十二週則有血管的增生，證實聚鄰甲氧基苯胺奈米絲薄膜不會引起慢性發炎反應。本研究初步證實聚甲氧基苯胺電紡絲薄膜屬於低毒性、良好生物相容性、低蛋白質吸附、低免疫抗原性；本材料雖然不利於纖維母細胞貼附，但卻適合肌肉細胞之貼附生長，此材料未來可能適合應用於肌肉組織工程。

關鍵字

生物相容性 ； 靜電紡絲 ； 聚鄰甲氧基苯胺奈米絲 ； 組織工程 ； 導電高分子

並列摘要

Abstract
Conducting polymers have both the physical and chemical characteristics of organic polymer as well as the conductivity of metals, which make them suitable to be used as photoelectric or biosensing materials. The purpose of the research was to study the biocompatibility of electrospinning poly o-methylaniline nanofiber and the feasibility for tissue engineering application. According to the SEM images, the diameter of poly o-methylaniline nanofiber was between 200 and 300 nm. Contact angle was 104 ± 8¬ o, which made its surface hydrophobic. Thermogravimetry analysis revealed that Tg was between 300 and 600 oC. The material had redox potential by cyclic voltammetry analysis. According to the ISO 10993-5 specification, we found that cytotoxicity of electrospinning poly o-methylaniline nanofiber was under the scope of acceptance prior and after electrical stimulation. From cell attachment and proliferation study, L929 fibroblasts had poor cell adhesion and proliferation on electrospinning poly o-methylaniline nanofiber compared with C2C12 myoblasts. By studying the gene expression of myoD, myogenin, myf-5, MRF4, myogenic differentiation of C2C12 cells was enhanced. In vitro biocompatibility studied by macrophages found that electrospinning poly o-methylaniline nanofiber induced the release of nitric oxide and inflammation cytokines. LDH release further demonstrated that the material caused some cell damage which elicited minor inflammation. Protein adsorption assay showed that human serum albumin and human plasma fibrinogen reached 80 μg/cm2 after 48 hr. In vivo biocompatibility was performed by implanting electrospinning poly o-methylaniline nanofiber in the back of Wistar rats subcutaneously. Some neutrophil and macrophages appeared after 1 and 4 weeks, and blood vessels were observed after 8 and 12 weeks. Our study suggested that electrospinning poly o-methylaniline nanofiber had relatively low toxicity, good biocompatibility, low protein adsorption and low immunogenicity. Although the material may not be suitable for fibroblast attachment, it is suitable for the attachment and growth of myoblasts. Electrospinning poly o-methylaniline nanofiber may be applied in muscle tissue engineering applications.

並列關鍵字

Biocompatibility ； Polyaniline nanofiber ； Electrospinning ； Conducting Polymer ； Tisssue Engineering

參考文獻

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