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神经元细胞与共价固定层粘连蛋白寡肽YIGSR和IKVAV的氟化乙烯丙烯膜的附着。II.

Neuronal cell attachment to fluorinated ethylene propylene films with covalently immobilized laminin oligopeptides YIGSR and IKVAV. II.

作者信息

Ranieri J P, Bellamkonda R, Bekos E J, Vargo T G, Gardella J A, Aebischer P

机构信息

Division of Surgical Research, Centre Hospitalier Universitaire Vaudois, Lausanne University Medical School, Switzerland.

出版信息

J Biomed Mater Res. 1995 Jun;29(6):779-85. doi: 10.1002/jbm.820290614.

Abstract

Material surfaces that can mediate cellular interactions by the coupling of specific cell membrane receptors may allow for the design of a biomaterial that can control cell attachment, differentiation, and tissue organization. Cell adhesion proteins have been shown to contain minimum oligopeptide sequences that are recognized by cell surface receptors and can be covalently immobilized on material surfaces. In this study, cell attachment to fluorinated ethylene propylene (FEP) films functionalized with the laminin-derived oligopeptides, YIGSR and a 19-mer IKVAV-containing sequence, was assessed using NG108-15 neuroblastoma and PC12 cells. A radiofrequency glow discharge (RFGD) process that replaces the FEP surface fluorine atoms with reactive hydroxyl functionalities was used to activate the film surfaces. The oligopeptides were then covalently coupled to the surface by their C-terminus using a standard nucleophilic substitution reaction. The covalent attachment of the oligopeptides to the FEP surface was verified using electron spectroscopy for chemical analysis (ESCA). Receptor-mediated NG108-15 cell attachment on the YIGSR-modified films was determined using competitive binding assays. Average cell attachment on the oligopeptide immobilized films in medium containing soluble CDPGYIGSR was reduced by approximately a factor of 2, compared to cell attachment in serum-free medium alone. No significant decrease in cell attachment was noted in medium containing the mock oligopeptide sequence CDPGYIGSK. FEP films immobilized with the 19-mer IKVAV sequence demonstrated a higher percentage of receptor mediated cell attachment on the film surfaces.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

能够通过特定细胞膜受体的偶联来介导细胞相互作用的材料表面,可能有助于设计一种能够控制细胞附着、分化和组织构建的生物材料。细胞黏附蛋白已被证明含有可被细胞表面受体识别的最小寡肽序列,并且可以共价固定在材料表面。在本研究中,使用NG108-15神经母细胞瘤细胞和PC12细胞评估了细胞对用层粘连蛋白衍生的寡肽YIGSR和含19个氨基酸的IKVAV序列功能化的氟化乙烯丙烯(FEP)膜的附着情况。采用射频辉光放电(RFGD)工艺,用反应性羟基官能团取代FEP表面的氟原子,以活化膜表面。然后通过标准亲核取代反应,将寡肽的C末端共价偶联到表面。使用化学分析电子能谱(ESCA)验证寡肽与FEP表面的共价连接。使用竞争性结合试验确定受体介导的NG108-15细胞在YIGSR修饰膜上的附着情况。与仅在无血清培养基中的细胞附着相比,在含有可溶性CDPGYIGSR的培养基中,寡肽固定膜上的平均细胞附着减少了约2倍。在含有模拟寡肽序列CDPGYIGSK的培养基中,未观察到细胞附着有显著下降。固定有19聚体IKVAV序列的FEP膜在膜表面显示出更高百分比的受体介导的细胞附着。(摘要截短于250字)

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