Li Rui, Song Xiao-ning, Zhang Hua, Dai Wen-bing, He Bing, Zheng Ying, Zhang Qiang, Wang Xue-qing
Yao Xue Xue Bao. 2016 Aug;51(8):1316-24.
The integrity of poly(ethylene glycol)-co-poly(ε-caprolactone) (PEG-PCL) micelles transcellular transported across madin-darby canine kidney(MDCK) epithelial cells was investigated. Fluorescein isothiocyanate isomer I(FITC) was conjugated to PEG-PCL and the product PEG-PCL-FITC was identified by fluorescence spectra. Two micelles were prepared using the thin-film hydration method: 3,3’-dioctadecyloxacarbocyanine perchlorate (DiO) and 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine perchlorate (DiI) co-loaded PEG-PCL micelles (DiO-DiI-M), DiI loaded and PEG-PCL-FITC contained micelles(FITC-DiI-M). The size of the micelles was characterized by dynamic light scattering analysis using a Malvern Zetasizer Nano ZS and it turned out that the particle sizes of both micelles were about 30 nm with identical polydispersity index(PDI). The stability of the micelles in phosphate buffer saline(PBS) was monitored using fluorescence spectra and both micelles were stable within 4 h in PBS. The integrity of PEG-PCL micelles in the transcellular process across MDCK epithelial cell monolayer at 1 and 4 h was investigated using laser confocal scanning microscope and Förster resonance energy transfer(FRET) technology. The Person’s coefficient and FRET efficiency of both Transwell layer and Receive layer were recorded. The results show that the FRET efficiency and Person’s coefficient of the Receive layer was consistent with that of Transwell layer for both the micelles at 1 h, but decreased at 4 h and FITC-DiI-M decreased more significantly than Di O-DiI-M. The results indicated that the micelles could transport across the MDCK monolayer intactly at 1 h but some of them were disassembled during the 4 h transportation process.
研究了聚（乙二醇）-共-聚（ε-己内酯）（PEG-PCL）胶束跨膜转运穿过麦氏犬肾（MDCK）上皮细胞的完整性。将异硫氰酸荧光素异构体I（FITC）与PEG-PCL偶联，并通过荧光光谱鉴定产物PEG-PCL-FITC。采用薄膜水化法制备了两种胶束：3,3'-二辛基氧杂碳菁高氯酸盐（DiO）和1,1'-二辛基-3,3,3',3'-四甲基吲哚碳菁高氯酸盐（DiI）共负载的PEG-PCL胶束（DiO-DiI-M）、负载DiI且含有PEG-PCL-FITC的胶束（FITC-DiI-M）。使用马尔文Zetasizer Nano ZS通过动态光散射分析对胶束的尺寸进行了表征，结果表明两种胶束的粒径均约为30 nm，多分散指数（PDI）相同。使用荧光光谱监测胶束在磷酸盐缓冲盐水（PBS）中的稳定性，两种胶束在PBS中4小时内均稳定。使用激光共聚焦扫描显微镜和Förster共振能量转移（FRET）技术研究了PEG-PCL胶束在1小时和4小时跨MDCK上皮细胞单层的跨细胞过程中的完整性。记录了Transwell层和接收层的皮尔逊系数和FRET效率。结果表明，两种胶束在1小时时接收层的FRET效率和皮尔逊系数与Transwell层一致，但在4小时时降低，且FITC-DiI-M降低得比DiO-DiI-M更显著。结果表明，胶束在1小时时可以完整地跨MDCK单层转运，但在4小时的转运过程中其中一些会解体。
