CIC biomaGUNE, San Sebastián, Gipuzkoa, Spain.
Macromol Biosci. 2013 Feb;13(2):234-41. doi: 10.1002/mabi.201200235. Epub 2013 Jan 11.
The intracellular delivery of Doxorubicin (Dox) from poly(lactide-co-glycolide) (PLGA) nanoparticles stabilised with bovine serum albumin, in HepG2 cells, is studied via flow cytometry, fluorescence lifetime imaging microscopy (FLIM), confocal Raman microscopy (CRM) and cell viability studies. Flow cytometry shows that the initial uptake of PLGA and Dox follow the same kinetics. However, following 8 h of incubation, the fluorescence intensity and cellular uptake of Dox decreases, while in the case of PLGA both parameters remain constant. FLIM shows the presence of a single-lifetime species, with a lifetime of 1.15 ns when measured inside the cells. Cell viability decreases by approximately 20% when incubated for 24 h with PLGA loaded with Dox, with a particle concentration of 100 µg · mL(-1). At the single-cell level, CRM shows changes in the bands from DNA and proteins in the cell nucleus when incubated with PLGA loaded with Dox.
通过流式细胞术、荧光寿命成像显微镜（FLIM）、共聚焦拉曼显微镜（CRM）和细胞活力研究，研究了载多柔比星（Dox）聚乳酸-共乙醇酸（PLGA）纳米颗粒在 HepG2 细胞中的细胞内递药情况，该纳米颗粒由牛血清白蛋白稳定。流式细胞术显示，PLGA 和 Dox 的初始摄取遵循相同的动力学。然而，孵育 8 小时后，Dox 的荧光强度和细胞摄取减少，而 PLGA 的这两个参数保持不变。FLIM 显示存在单一寿命物种，在细胞内测量时寿命为 1.15 ns。当用浓度为 100 µg · mL(-1) 的载有 Dox 的 PLGA 孵育 24 小时时，细胞活力下降约 20%。在单细胞水平上，当用载有 Dox 的 PLGA 孵育时，CRM 显示细胞核中 DNA 和蛋白质的谱带发生变化。
