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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (7): 32-37    
研究报告     
材料表面化学刺激对纤连蛋白构象及细胞的影响
霍润兰, 罗彦凤, 李岩, 严维维, 邢娟, 林福春, 王远亮
生物流变科学与技术教育部重点实验室 重庆大学生物工程学院 重庆 400030
Surface Chemistry Modulates Fibronectin Conformation and Control Cell Behavior
HUO Run-lan, LUO Yan-feng, LI Yan, YAN Wei-wei, XING Juan, LIN Fu-chun, WANG Yuan-liang
Key Laboratory of Biorheological Science and Technology, Bioenginering College of Chongqing University, Research Center of Bioinspired Material Science and Engineering, Chongqing 400030, China
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摘要:

生物材料的表面化学性质通过改变吸附蛋白的构象,影响细胞粘附铺展,产生不同的细胞行为。采用金-硫自组装单分子层技术(alkanethiol self-assembled monolayers,SAMs)构建了不同化学基团(-NH2、-COOH、-OH和-CH3)修饰的基底材料表面。运用X射线光能质谱(XPS)和接触角仪表征材料表面化学组分以及亲疏水性,通过免疫荧光法测定材料表面预吸附蛋白—纤连蛋白的构象, 同时,应用MTT法检测细胞在基质材料上第1、3、5、7、9、11、13天的增殖活力,观察了成骨细胞铺展的影响。实验结果表明,亲水性材料表面(-OH与-COOH)对纤连蛋白构象影响较低,疏水性材料表面(-CH3)影响最大,中性材料表面(-NH2)介于二者之间。成骨细胞粘附和增殖能力也与蛋白质构象变化相关。

关键词: 自组装单分子层纤连蛋白构象细胞增殖细胞铺展    
Abstract:

Biomaterial surface properties modulate protein adsorption and cell adhesion, proliferation, spreading to elicit diverse cellular responses in biomedical and biotechnological applications. Alkanethiol self-assembled monolayers (SAMs) presenting well-defined chemistries (NH2, COOH, OH and CH3) were employed to analyze the effects of surface chemistry on controlling fibronectin (FN) conformation as well as osteoblasts behaviors. X-ray photoelectron spectroscopy (XPS) is used to confirm the presence of well-formed monolayers of expected physicochemical characteristics, and the hydrophilicity of SAMs was characterized by water contact angle. Immunostaining were applied to indicate conformation of FN. In addition, the effects of SAMs with pre-coated FN on the cytoskeleton organization and proliferation were also examined. Water contact angle data reveals the-CH3groups lead to a hydrophobic surface, while COOH, OH groups lead to a hydrophilic surface and NH2groups a neutral surface. SAMs presenting terminal NH2, COOH, OH and CH3 functionalities modulated adsorbed FN conformation as determined through differences in the binding affinities of monoclonal antibodies raised against the central cell-binding domain,-OH surfaces exhibited a statistically greater HFN 7.1 positive area than the other surfaces. Then, the changes in osteoblasts proliferation and spreading correlated with the FN conformation.

Key words: Self-assembled monolayers    Fibronectin    Conformation    Proliferation    Spreading
收稿日期: 2011-03-04 出版日期: 2011-07-25
ZTFLH:  Q2  
基金资助:

国家自然科学基金(30760251)、教育部博士点基金(20070678002)、云南省科技计划(2009CA005)资助项目

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王远亮
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引用本文:

霍润兰, 罗彦凤, 李岩, 严维维, 邢娟, 林福春, 王远亮. 材料表面化学刺激对纤连蛋白构象及细胞的影响[J]. 中国生物工程杂志, 2011, 31(7): 32-37.

HUO Run-lan, LUO Yan-feng, LI Yan, YAN Wei-wei, XING Juan, LIN Fu-chun, WANG Yuan-liang. Surface Chemistry Modulates Fibronectin Conformation and Control Cell Behavior. China Biotechnology, 2011, 31(7): 32-37.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I7/32


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