Fig. 5

Targeting capacity of the TopFluor® (TF) labelled VitE:SM nanoemulsions in vitro and in vivo. a TopFluor® was incorporated in the surfactant (TopFluor®-labelled-sphingomyelin) and injected in the aqueous phase as previously explained (Fig. 1a). VitE:SM:TF = Vitamin E + TopFluor®-labelled-sphingomyelin nanosystem. b Confocal images of muBMDM cells treated or not with TF-labelled nanoemulsions (VitE:SM:TF, 0.5 mg/mL). Scale bar = 20 µM. DAPI: blue, TF: green. c Cytometry dot plots of muIBMDM cells in the presence or absence of VitE:SM:TF (0.5 mg/mL) nanoemulsions. Murine IBMDMs containing VitE:SM:TF nanoemulsions were detected at a wavelength of Ex488 nm Em530/30 nm. SSC-A: Side Scatter (Area). d Flow cytometry analysis of the percentage of TopFluor®-positive live cells ± SD in three different organs (liver, tumor, and lung) under two treatment modalities: intraperitoneal or retro-orbital (n = 2 mice per treatment). e Flow cytometry analysis of the percentage of TopFluor®-positive live cells ± SD in the liver under two intravenous treatment modalities (tail vein injection or retro-orbital) compared to control (n = 5 control, n = 5 mice per each treatment). ∗  = p < 0.05; ∗  ∗  ∗  = p < 0.001. One-way ANOVA with Dunnett’s post hoc test, compared to the retro-orbital condition. f Schematic representation of the markers used to identify macrophages: CD45 + , CD11b + and F4/80 + . g Flow cytometry analysis of the percentage of macrophages (CD45 + , CD11b + and F480 +) ± SD targeted by the VitE:SM:TF nanoemulsions over time, with data collected at 1 h, 4 h, 24 h and 72 h. Unpaired t test comparing 24 and 72 h. ns  not significant