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支持细胞模拟单层及其与血液的相互作用。

Supported cell mimetic monolayers and their interaction with blood.

作者信息

Kaladhar K, Sharma Chandra P

机构信息

Biosurface Technology Division, BMT Wing, Sree Chithra Tirunal Institute for Medical Science and Technology, Poojappura 695012, Thiruvananthapuram, Kerala, India.

出版信息

Langmuir. 2004 Dec 7;20(25):11115-22. doi: 10.1021/la048644y.

Abstract

Surface modification using supported monolayers of phosphorylcholine containing phospholipids has been an accepted strategy for developing blood-contacting materials. We present a detailed study of the blood compatibility of the supported monolayers of phospholipid, glycolipid, and cholesterol (Chol) binary and ternary lipid combinations using in vitro techniques. The packing and orientation of these monolayers have been correlated with the blood compatibility. We have used phosphatidylcholine (PTC) for phospholipid, galactocerebroside (Gal) for glycolipid, and Chol based on the headgroup structure to represent the major lipid components of the endothelial luminal cell membrane. The interfacial behavior of various combinations of PTC, Gal, and Chol monolayers have been studied at the air/water interface and deposited on hydrophobic polycarbonate (PC) polymer substrates with the help of the Langmuir-Blodgett trough. The packing and orientation of the supported monolayers have been varied by means of changing the lipid composition rather than the deposition parameters. This approach seems to be more similar to the in vivo conditions. The different supported monolayer surfaces prepared accordingly are (1) a closely packed ordered hydrophobic surface, PC modified with the combination PTC/Chol/Gal (1:0.35:0.125), (2) a loosely packed ordered hydrophobic surface, PC modified with the combination PTC/Chol (1:0.35), and (3) a closely packed ordered hydrophilic surface, PC modified with the combination PTC/Chol (1:0.7). An optimized modified surface (PTC/Chol/Gal, 1:0.35:0.125) has been identified on the basis of the maximum transfer ratio from the air/water interface and characterized by using atomic force microscopy. The concentration of Chol has been found to be an important parameter, which influences the transfer ratio. The Gal improves the monolayer integrity under a reduced Chol concentration. The blood compatibility of these supported monolayers was studied by protein adsorption, blood cell adhesion, and calcification. The tightly packed ordered hydrophobic surface (PTC/Chol/Gal, 1:0.35:0.125), has been found to be more blood compatible because of reduced blood cell adhesion and calcification. This surface also promotes albumin adsorption and may be the reason for the reduced platelet activation, while in the case of the loosely packed ordered hydrophobic surface (PTC/Chol, 1:0.35) the protein adsorption also has been reduced along with the blood cell adhesion and calcification. When the ordered hydrophilic surface (PTC/Chol, 1: 0.7) of the monolayer has been exposed, the blood cell adhesions as well as the overall protein adsorption were significantly reduced. However, the packing of the phosphorylcholine moieties of the polar headgroup has been affecting the calcification on the surface. We have observed an increase in calcification to the surface modified with the loosely packed polar headgroup, from a relative study on chitosan and chitosan modified with the monolayer of PTC. These findings are helpful for the surface modifications for blood-contacting materials using this strategy.

摘要

使用含磷酰胆碱的磷脂支撑单分子层进行表面改性一直是开发血液接触材料的一种公认策略。我们使用体外技术对磷脂、糖脂和胆固醇(Chol)二元和三元脂质组合的支撑单分子层的血液相容性进行了详细研究。这些单分子层的堆积和取向与血液相容性相关。基于头基结构,我们使用磷脂酰胆碱(PTC)作为磷脂,半乳糖脑苷脂(Gal)作为糖脂,以及Chol来代表内皮腔细胞膜的主要脂质成分。在空气/水界面研究了PTC、Gal和Chol单分子层各种组合的界面行为,并借助朗缪尔-布洛杰特槽将其沉积在疏水性聚碳酸酯(PC)聚合物基底上。通过改变脂质组成而非沉积参数来改变支撑单分子层的堆积和取向。这种方法似乎更类似于体内条件。据此制备的不同支撑单分子层表面有:(1)紧密堆积的有序疏水表面,用PTC/Chol/Gal(1:0.35:0.125)组合改性的PC;(2)松散堆积的有序疏水表面,用PTC/Chol(1:0.35)组合改性的PC;(3)紧密堆积的有序亲水表面,用PTC/Chol(1:0.7)组合改性的PC。基于从空气/水界面的最大转移率确定了一个优化的改性表面(PTC/Chol/Gal,1:0.35:0.125),并使用原子力显微镜对其进行了表征。已发现Chol的浓度是一个重要参数,它会影响转移率。在降低的Chol浓度下,Gal可改善单分子层的完整性。通过蛋白质吸附、血细胞粘附和钙化研究了这些支撑单分子层的血液相容性。已发现紧密堆积的有序疏水表面(PTC/Chol/Gal,1:0.35:0.125)具有更高的血液相容性,因为血细胞粘附和钙化减少。该表面还促进白蛋白吸附,这可能是血小板活化降低的原因,而在松散堆积的有序疏水表面(PTC/Chol,1:0.35)的情况下,蛋白质吸附以及血细胞粘附和钙化也有所减少。当单分子层的有序亲水表面(PTC/Chol,1:0.7)暴露时,血细胞粘附以及总体蛋白质吸附显著减少。然而,极性头基的磷酰胆碱部分的堆积会影响表面的钙化。通过对壳聚糖和用PTC单分子层改性的壳聚糖的相对研究,我们观察到用松散堆积的极性头基改性的表面钙化增加。这些发现有助于使用该策略对血液接触材料进行表面改性。

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