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聚(四氟乙烯 - 共 - 六氟丙烯)的肽表面修饰增强了其与中枢神经系统神经元的相互作用。

Peptide surface modification of poly(tetrafluoroethylene-co-hexafluoropropylene) enhances its interaction with central nervous system neurons.

作者信息

Tong Y W, Shoichet M S

机构信息

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ontario, Canada.

出版信息

J Biomed Mater Res. 1998 Oct;42(1):85-95. doi: 10.1002/(sici)1097-4636(199810)42:1<85::aid-jbm11>3.0.co;2-n.

Abstract

Poly(tetrafluoroethylene-co-hexafluoropropylene) (FEP) film surfaces were chemically surface modified to introduce one of three laminin adhesive peptides: GYIGSR, GRGDS, or SIKVAV. FEP film surfaces were first reduced with sodium naphthalide to introduce surface carbon-carbon double bonds at two reaction conditions: 20 min at -78 degrees C, and 3 h at 25 degrees C. Scanning electron microscopy and atomic force microscopy indicated that surface topography was unaffected by the reaction conditions. Reduced FEP film surfaces were further modified to introduce hydroxyl groups via hydroboration/oxidation or carboxylic acid groups via oxidation. The hydroxyl (FEP-CHxOH) and carboxylic acid (FEP-COOH) functionalized surfaces provided reactive handles for peptide coupling using tresyl chloride. Surface elemental composition data, determined from X-ray protoelectron spectroscopy, indicated that equivalent amounts of GYIGSR, GRGDS, and SIKVAV were introduced. Two additional coupling reagents, SMCC and TSU, were compared to tresyl chloride for the coupling of radio-labeled tyrosine of GYIGSR. Between 8 and 150 fmol/cm2 of peptide was introduced to the hydroxyl and carboxylic acid functionalized surfaces, with the tresyl coupling reagent showing the greatest amount of peptide incorporated. The tresyl-coupled peptide-modified surfaces were compared in terms of the response of primary, embryonic hippocampal neurons plated from serum-free medium for 4 days. The number and length of neurites extending from the cell bodies were averaged over 50 cells after 1 and 4 days FEP-CHxO-peptide surfaces had either a greater or equivalent hippocampal neuron interaction than the corresponding FEP-COO-peptide surfaces. All peptide-functionalized surfaces had a greater hippocampal neuron interaction than the corresponding FEP-CHxOH, FEP-COOH, and FEP controls after 4 days underlying the importance of the peptides over hydrophilic or hydrophobic surfaces. After 4 days differences in neurite extension were evident among the peptide-functionalized surfaces, with the longest neurites observed on the SIKVAV-functionalized surfaces.

摘要

聚(四氟乙烯 - 共 - 六氟丙烯)(FEP)薄膜表面经过化学表面改性,以引入三种层粘连蛋白粘附肽之一:GYIGSR、GRGDS或SIKVAV。FEP薄膜表面首先用萘钠还原,在两种反应条件下引入表面碳 - 碳双键:在-78℃下反应20分钟,以及在25℃下反应3小时。扫描电子显微镜和原子力显微镜表明,表面形貌不受反应条件的影响。还原后的FEP薄膜表面通过硼氢化/氧化进一步改性以引入羟基,或通过氧化引入羧酸基团。羟基(FEP-CHxOH)和羧酸(FEP-COOH)功能化表面提供了用于使用三甲苯磺酰氯进行肽偶联的反应性基团。由X射线光电子能谱测定的表面元素组成数据表明,引入了等量的GYIGSR、GRGDS和SIKVAV。比较了另外两种偶联试剂SMCC和TSU与三甲苯磺酰氯用于GYIGSR的放射性标记酪氨酸的偶联。在羟基和羧酸功能化表面上引入了8至150 fmol/cm² 的肽,其中三甲苯磺酰偶联试剂显示掺入的肽量最大。比较了三甲苯磺酰偶联肽修饰表面在无血清培养基中培养4天的原代胚胎海马神经元的反应。在1天和4天后,从细胞体延伸的神经突的数量和长度在50个细胞上进行平均,FEP-CHxO-肽表面比相应的FEP-COO-肽表面具有更大或相当的海马神经元相互作用。4天后,所有肽功能化表面比相应的FEP-CHxOH、FEP-COOH和FEP对照具有更大的海马神经元相互作用,这突出了肽相对于亲水或疏水表面的重要性。4天后,肽功能化表面之间神经突延伸的差异很明显,在SIKVAV功能化表面上观察到最长的神经突。

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