Endothelial glycocalyx conditions influence nanoparticle uptake for passive targeting.
Endothelial glycocalyx conditions influence nanoparticle uptake for passive targeting.
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DOI：
10.2147/ijn.s106299
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发表时间：
2016
影响因子：
8
通讯作者：
Ebong EE
中科院分区：
文献类型：
--
作者：
Cheng MJ;Kumar R;Sridhar S;Webster TJ;Ebong EE
心血管疾病是由内皮细胞(EC)功能障碍引起的，并与EC糖皮脱落相吻合。这些疾病可以通过使用循环纳米粒(NP)药物载体将药物输送到受影响的血管区域来预防。本研究的目的是观察10 nm聚乙二醇包覆金纳米颗粒(PEGAuNP)在ECs中的传递情况，以及EC表面的糖基化结构对其的影响。选择大鼠脂肪垫内皮细胞，通过吸附白蛋白和整合硫酸乙酰肝素(HS)链的荧光免疫标记，证实其具有健壮的糖基化反应。共聚焦荧光成像显示约3微米厚的糖帽层，覆盖75%的内皮细胞并含有丰富的HS。正如预期的那样，这种健康的糖萼阻碍了对聚乙二醇AuNP的吸收，因为糖萼的毛孔通常为7纳米宽。其他被测试的糖萼模型包括：通过在降低蛋白质的培养液中培养细胞而获得的崩溃的糖萼，通过应用肝素酶III酶特异性地切割HS而获得的降解的糖萼，以及通过在酶降解后将外源HS添加到培养基中而获得的恢复的糖萼。折叠的Waŝ2微米厚，EC覆盖率不变，HS含量持续不变。降解的糖萼在EC厚度和覆盖率方面表现出类似的变化，但其HS厚度减少到0.7微米，仅覆盖原始EC表面的10%。两种功能障碍的模型保留的聚乙二醇AuNP比健康的糖萼多六到七倍。塌陷的糖萼允许NPs穿过糖萼进入细胞内空间，而降解的糖萼将聚乙二醇AuNP捕获在糖萼内。修复后的糖萼模型部分恢复了HS的厚度为1.2微米，内皮细胞覆盖率为44%，但它能够将NP摄取恢复到基线水平。综上所述，这项研究表明，糖萼结构是内皮细胞摄取NP的关键，可能是将NPs运送到功能障碍的ECs的被动途径。
Cardiovascular diseases are facilitated by endothelial cell (EC) dysfunction and coincide with EC glycocalyx coat shedding. These diseases may be prevented by delivering medications to affected vascular regions using circulating nanoparticle (NP) drug carriers. The objective of the present study was to observe how the delivery of 10 nm polyethylene glycol-coated gold NPs (PEG-AuNP) to ECs is impacted by glycocalyx structure on the EC surface. Rat fat pad endothelial cells were chosen for their robust glycocalyx, verified by fluorescent immunolabeling of adsorbed albumin and integrated heparan sulfate (HS) chains. Confocal fluorescent imaging revealed a ~3 µm thick glycocalyx layer, covering 75% of the ECs and containing abundant HS. This healthy glycocalyx hindered the uptake of PEG-AuNP as expected because glycocalyx pores are typically 7 nm wide. Additional glycocalyx models tested included: a collapsed glycocalyx obtained by culturing cells in reduced protein media, a degraded glycocalyx obtained by applying heparinase III enzyme to specifically cleave HS, and a recovered glycocalyx obtained by supplementing exogenous HS into the media after enzyme degradation. The collapsed glycocalyx waŝ2 µm thick with unchanged EC coverage and sustained HS content. The degraded glycocalyx showed similar changes in EC thickness and coverage but its HS thickness was reduced to 0.7 µm and spanned only 10% of the original EC surface. Both dysfunctional models retained six- to sevenfold more PEG-AuNP compared to the healthy glycocalyx. The collapsed glycocalyx permitted NPs to cross the glycocalyx into intracellular spaces, whereas the degraded glycocalyx trapped the PEG-AuNP within the glycocalyx. The repaired glycocalyx model partially restored HS thickness to 1.2 µm and 44% coverage of the ECs, but it was able to reverse the NP uptake back to baseline levels. In summary, this study showed that the glycocalyx structure is critical for NP uptake by ECs and may serve as a passive pathway for delivering NPs to dysfunctional ECs.