J. Mater. Sci. Technol. ›› 2010, Vol. 26 ›› Issue (12): 1119-1126.

• Research Articles • 上一篇    下一篇

一种温度敏感角膜上皮细胞培养材料的合成

关燕清1,刘俊明2,王馨3,倪小丽3,杨爱妮3   

  1. 1. 华南师范大学生命科学学院
    2. 南京大学
    3. 华南师范大学
  • 收稿日期:2009-10-14 修回日期:2010-01-20 出版日期:2010-12-31 发布日期:2010-12-21
  • 通讯作者: 关燕清

Synthesis of a Kind of Temperature-responsive Cell Culture Surface for Corneal Sheet

Yanqing Guan, Zhibin Li, Xin Wang, Xiaoli Ni, Aini Yang, Junming Liu   

  1. 1) School of Life Science, South China Normal University, Guangzhou 510631, China
    2) School of Physics, South China Normal University, Guangzhou 510631, China
    3) Laboratory of Solid State Microstructures, Nanjing University, Nanjing 210093, China
    4) International Center for Materials Physics, Chinese Academy of Sciences, Shenyang 110016, China
  • Received:2009-10-14 Revised:2010-01-20 Online:2010-12-31 Published:2010-12-21
  • Contact: GUAN Yan-Qing
  • Supported by:

    the National Natural Science Foundation of China (Nos. 30970731 and 50832002), the Natural Science Foundation of Guangdong Province (No. 9151063101000015). The authors thank Professor Zhicong Wang (State Key Laboratory of Ophthalmology, Guangzhou) for kindly providing corneal epithelial cells and thank Professors Qili Fen and Changlian Peng for useful comments.

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摘要： 用丙烯酸(AAc)对N-异丙基丙烯酰胺(NIPAAm)进行化学改性，合成反应中间体PIA，进一步与叠氮苯甲酸(4-Azidoaniline)合成光活性温度应答高分子AzPhPIA。通过高效液相色谱和红外光谱表征产物的结构和组成，并通过降温实验和浊度检测研究产物的温敏性。结果表明AzPhPIA共聚物的相转变温度(LCST)为21.5°C，比NIPAAm的LCST降低近10°C，具有更广泛的组织工程临床应用前景。将光活性的AzPhPIA共聚物以紫外光化学接枝法固定化于聚苯乙烯组织培养板表面并对该培养板进行细胞培养研究。结果表明，AzPhPIA固定化的聚苯乙烯组织培养板具有良好的细胞相容性。37°C时，人子宫颈癌细胞(HeLa)贴附生长情况良好。当环境温度降低至15°C时，AzPhPIA共聚物发生疏水-亲水变化，导致培养板表面的细胞层自动脱附。这种免载体细胞培养技术不仅避免了使用酶解法脱附细胞造成的细胞功能损伤，同时克服了现有组织工程角膜支架材料在体内降解引发移植排斥和并发症的难题，有着广阔的应用前景。

关键词: N-聚异丙基丙烯酰胺, 相转变温度, 组织工程, 角膜上皮细胞

Abstract: In this study, acrylic acid (AA) and 4-azidoaniline were used to modify poly (N-isopropylacrylamide) (NI-PAAm) in order to fabricate temperature-responsive surface for corneal epithelia cell adhesion and detachment. First, NIPAAm was copolymerized with acrylic acid. Then, the copolymer was coupled with azidoaniline to synthesize AzPhPIA, derivative of p(NIPAAm-co-AA), which possesses both thermo- and photo-sensitivities. Second, the synthesized copolymer was characterized by high performance liquid chromatography (HPLC), Fourier transform infrared (FTIR) and a CHN analyzer. The thermo-sensitivity was characterized by temperature reducing experiment, contact angle measurement and low critical solution temperature (LCST) testing. Third, the derivatized copolymer was immobilized by photolithography on a polystyrene plate, and then the surface characterization of AzPhPIA-coated polystyrene plate (PSt) was measured by electron spectroscopy for chemical analysis (ESCA). The thermo-sensitivity and cytocompatibility of the AzPhPIA-coated PSt were investigated by corneal epithelial cells culture. The results revealed that the AzPhPIA-coated PSt exhibited good cytocompatibility and cell detachability when temperature decreased.

Key words: N-isopropylacrylamide, Low critical solution temperature, Tissue engineering, Corneal epithelial cells