Preferential binding of polyethylene glycol-coated liposomes containing a novel cationic lipid, TRX-20, to human subendthelial cells via chondroitin sulfate
Preferential binding of polyethylene glycol-coated liposomes containing a novel cationic lipid, TRX-20, to human subendthelial cells via chondroitin sulfate
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DOI：
10.1023/a:1013033826974
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发表时间：
2001-09-01
影响因子：
3.7
通讯作者：
Kimura, J
中科院分区：
文献类型：
--
作者：
Harigai, T;Kondo, M;Kimura, J
目的。为了设计新型阳离子脂质体，制备了含有新合成的阳离子脂类3，5-二十五烷氧基苯甲酰胺盐酸盐(TRX-20)的聚乙二醇包被的阳离子脂质体，并研究了其在人血管内皮细胞(主动脉平滑肌细胞和系膜细胞)和人内皮细胞中的体外结合和摄取。采用挤出法制备了3种不同的聚乙二醇包被阳离子脂质体，并测定了它们的平均粒径和Zeta电位。将罗丹明标记的聚乙二醇包被的阳离子脂质体与血管、肾小球系膜细胞和血管内皮细胞在37℃下孵育24 h，通过测量细胞相关的荧光强度来估计脂质体的细胞结合量和摄取量。为了考察脂质体的结合特性，考察了不同类型的糖胺聚糖裂解酶对细胞结合的变化。荧光显微镜用于寻找脂质体在细胞中的定位。聚乙二醇包被的阳离子脂质体与平滑肌细胞的结合和摄取强烈依赖于阳离子脂质体中阳离子脂类的化学形态。与其他含有N-(1-(2，3-二油酰氧基)丙基)-N，N，N-三甲基铵盐或N-(alpha-(trimethylammonio)acetyl)-D-glutamate氯化物的阳离子脂质体相比，血管细胞与聚乙二醇包被的Trx-20脂质体的结合量更高。尽管聚乙二醇包被的TRX-20脂质体对内皮下细胞具有较高的亲和力，但它们与内皮细胞的结合很小。某些软骨素酶可抑制细胞与内皮下细胞的结合，而肝素酶不能抑制其与内皮细胞的结合。这些结果表明，聚乙二醇包被的TRX-20脂质体通过与细胞表面和细胞外基质中的某些硫酸软骨素蛋白多糖(不与硫酸乙酰肝素蛋白多糖)相互作用，对内皮下细胞具有很强的选择性结合性能。这种结合特性不同于已报道的其他阳离子脂质体。聚乙二醇包被的TRX-20脂质体可以通过某些类型的硫酸软骨素蛋白多糖与内皮下细胞强烈和选择性地结合，作为一种特异的药物输送系统将具有优势。
Purpose. To design novel cationic liposomes, polyethylene glycol (PEG)-coated cationic liposomes containing a newly synthesized cationic lipid, 3,5-dipentadecyloxybenzamidine hydrochloride (TRX-20) were formulated and their cellular binding and uptake investigated in vitro in the following cells: human subendothelial cells (aortic smooth muscle cells and mesangial cells) and human endothelial cells.Methods. Three different PEG-coated cationic liposomes were prepared by the extrusion method, and their mean particle size and zeta potential were determined. Rhodamine-labeled PEG-coated cationic liposomes were incubated with smooth muscle cells, mesangial cells, and endothelial cells at 37 degreesC for 24 h. The amounts of cellular binding and uptake of liposomes were estimated by measuring the cell-associated fluorescence intensity of rhodamine. To investigate the binding property of the liposomes, the changes of the binding to the cells pretreated by various kinds of glycosaminoglycan lyases were examined. Fluorescence microscopy is used to seek localization of liposomes in the cells.Results. The cellular binding and uptake of PEG-coated cationic liposomes to smooth muscle cells was depended strongly on the chemical species of cationic lipids in these liposomes. Smooth muscle cells bound higher amount of PEG-coated TRX-20 liposomes than other cationic liposomes containing N-(1-(2,3-dioleoyloxy) propyl)-N, N, N-trimethylammonium salts or N-(alpha-(trimethylammonio)acetyl)-D-glutamate chloride. Despite of the higher affinity of PEG-coated TRX-20 liposomes for subendothelial cells, their binding to endothelial cells was very small. The binding to subendothelial cells was inhibited when cells were pretreated by certain kinds of chondroitinase, but not by heparitinase. These results suggest that PEG-coated TRX-20 liposomes have strong and selective binding property to subendothelial cells by interacting with certain kinds of chondroitin sulfate proteoglycans (not with heparan sulfate proteoglycans) on the cell surface and in the extracellular matrix of the cells. This binding feature was different from that reported for other cationic liposomes.Conclusions. PEG-coated TRX-20 liposomes can strongly and selectively bind to subendothelial cells via certain kinds of chondroitin sulfate proteoglycans and would have an advantage to use as a specific drug delivery system.