Study of Intracellular Delivery of Doxorubicin from Poly(lactide-co-glycolide) Nanoparticles by Means of Fluorescence Lifetime Imaging and Confocal Raman Microscopy

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 8h 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 1.15ns when measured inside the cells. Cell viability decreases by approximately 20% when incubated for 24h with PLGA loaded with Dox, with a particle concentration of 100 mu g center dot mL1. 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.