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通过肽键在聚合物表面实现外细胞模拟双层膜的细胞模拟侧向稳定及其血液相容性。

Cell mimetic lateral stabilization of outer cell mimetic bilayer on polymer surfaces by peptide bonding and their blood compatibility.

作者信息

Kaladhar K, Sharma Chandra P

机构信息

Biosurface Technology Division, BMT Wing, Sree Chithira Tirunal Institute for Medical Science and Technology, Thiruvananthapuram, Kerala, India.

出版信息

J Biomed Mater Res A. 2006 Oct;79(1):23-35. doi: 10.1002/jbm.a.30681.

Abstract

The biological lipid bilayer membranes are stabilized laterally with the help of integral proteins. We have simulated this with an optimized ternary phospholipid/glycolipid/cholesterol system, and stabilized laterally on functionalized poly methyl methacrylate (PMMA) surfaces, using albumin, heparin, and polyethylene glycol as anchors. We have earlier demonstrated the differences due to orientation and packing of the ternary phospholipid monolayers in relation to blood compatibility (Kaladhar and Sharma, Langmuir 2004;20:11115-11122). The structure of albumin is changed here to expose its interior hydrophobic core by treating with organic solvent. The interaction between the hydrophobic core of the albumin molecule and the hydrophobic core of the lipid molecules is confirmed by incorporating the molecule into bilayer membranes. The secondary structure of the membrane incorporated albumin is studied by CD spectral analysis. The structure of the altered albumin molecule contains more beta-sheet as compared to the native albumin. This conformation is also retained in membranes. The partitioning of the different anchors based on its polarity and ionic interactions in the monolayer is studied from the pressure-area (pi-A) isotherm of the lipid monolayers at the air/water interface using Langmuir-Blodgett (LB) trough facility. Such two monolayers are deposited onto the functionalized PMMA surface using LB trough and crosslinked by carbodiimide chemistry. The structure of the deposited bilayer is studied by depth analysis using contact mode AFM in dry conditions. The stabilized bilayer shows stability up to 1 month by contact angle studies. Preliminary blood compatibility studies reveal that the calcification, protein adsorption, as well as blood-cell adhesion is significantly reduced after the surface modification. The reduced adsorption of ions, proteins, and cells to the modified surfaces may be due to the fluidity of the microenvironment along with the contribution of the mobile PEG groups at the surface and the phosphorylcholine groups of the phospholipids. The stability of the anchored bilayer under low shear stress conditions promises that the laterally stabilized supported bilayer system can be used for low shear applications like small diameter vascular graft and modification of biosensors, and so forth.

摘要

生物脂质双分子层膜在整合蛋白的帮助下实现横向稳定。我们使用优化的三元磷脂/糖脂/胆固醇体系对此进行了模拟,并以白蛋白、肝素和聚乙二醇作为锚定物,在功能化的聚甲基丙烯酸甲酯(PMMA)表面实现了横向稳定。我们之前已经证明了三元磷脂单层的取向和堆积与血液相容性之间的差异(Kaladhar和Sharma,《朗缪尔》2004年;20:11115 - 11122)。此处通过用有机溶剂处理改变白蛋白的结构,以暴露其内部疏水核心。通过将该分子整合到双分子层膜中,证实了白蛋白分子的疏水核心与脂质分子的疏水核心之间的相互作用。通过圆二色光谱分析研究了膜结合白蛋白的二级结构。与天然白蛋白相比,改变后的白蛋白分子结构含有更多的β - 折叠。这种构象在膜中也得以保留。利用Langmuir - Blodgett(LB)槽装置,从空气/水界面处脂质单层的压力 - 面积(π - A)等温线研究了基于其极性和离子相互作用的不同锚定物在单层中的分配情况。使用LB槽将这样的两个单层沉积到功能化的PMMA表面,并通过碳二亚胺化学交联。在干燥条件下,使用接触模式原子力显微镜通过深度分析研究沉积双分子层的结构。通过接触角研究表明,稳定的双分子层在长达1个月的时间内保持稳定。初步血液相容性研究表明,表面改性后钙化、蛋白质吸附以及血细胞黏附显著减少。离子、蛋白质和细胞对改性表面吸附的减少可能归因于微环境的流动性以及表面移动的PEG基团和磷脂的磷酰胆碱基团的贡献。锚定双分子层在低剪切应力条件下的稳定性保证了横向稳定的支撑双分子层系统可用于小直径血管移植物和生物传感器改性等低剪切应用场景。

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