PMC

Physiochemical characterization of Nufs-DTX.
Notes: (A) A simple schematic of generation of Nufs-DTX. (B) The Nufs-DTX dispersion solution in DI water and size distribution by Zetasizer. (C) The physical stability of Nufs-DTX for 2 months at room temperature.
Abbreviations: DI, deionized; DTX, docetaxel; Nufs, nanoparticulation using fat and supercritical fluid; PVP, polyvinylpyrrolidone; PDI, polydispersity index.

In vivo distribution and tumor growth delay of Nufs-DTX.
Notes: (A) Tissue distribution of Nufs-DTX after intravenous injection. Unlike Taxotere™ that was accumulated mainly in the lungs after intravenous administration, Nufs-DTX was observed mainly in the liver. Taxotere™ was detected in the main organs including the lungs, liver, spleen, and kidneys over the collecting time points (hour), while Nufs-DTX was well cleared from each organ. (B) In serum, both the drugs showed similar pattern in distribution and clearance over a period of time. In tumors, a relatively large amount of accumulated Nufs-DTX was observed compared with Taxotere™. (C) Tumor growth delay after injecting three times every other day. Nufs-DTX showed effective tumor growth delay compared with other controls, including Taxotere™-treated group. Arrows indicate time of injections.
Abbreviations: DTX, docetaxel; Nufs, nanoparticulation using fat and supercritical fluid.

Cellular response of Nufs-DTX.
Notes: (A) Polymerization of alpha-tubulin after drug treatment. After treatment of A549 cells with Nufs-DTX for 16 hours, excess polymerization of microtubules (green fluorescence) was observed in the cytoplasm, as observed with Taxotere™ treatment. DAPI staining indicates nuclei. Scale bar: 20 μm. (B) G2/M arrest after drug treatment. Unlike control, cells treated with Taxotere™ or Nufs-DTX showed G2/M arrest for at least 72 hours by flow cytometry analysis. (C) The proliferation rate after drug treatment for an acute response. There was no difference in the percentage of Ki-67-positive cells at the same concentrations between Taxotere™-treated cells and Nufs-DTX-treated cells. (D) Clonogenic assay after drug treatment for a chronic response. There was no difference in the survival fraction at the same concentrations between Taxotere™-treated cells and Nufs-DTX-treated cells.
Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; DTX, docetaxel; Nufs, nanoparticulation using fat and supercritical fluid.

In vivo toxicity of Nufs-DTX.
Notes: (A) All values of CBC count (WBCs, neutrophils, and platelets) of both the groups were within normal ranges. (B) All values of AST, ALT, BUN, and creatinine in both the groups were within normal ranges. (C) Kaplan–Meier plot (n=10), right panel shows changes in body weight (P<0.001, Student’s t-test between the Taxotere™ group and the Nufs-DTX group). While there was no event of death observed in the Nufs-DTX-treated group, only 20% survival rate was observed in the Taxotere™ 30 mg/kg group. A significant loss of body weight was observed in a Taxotere™-treated group. (D) Paw withdrawal latency time (n=5). Taxotere™ significantly increased thermal threshold (*P<0.05, Student’s t-test between the Taxotere™ group and the control group). (E) An electron microscopic analysis of neuropathological changes in the sciatic nerve after Nufs-DTX treatment. While there were no or minor myelin shape changes in the Nufs-DTX-treated group, collapsed or irregular axons and abnormally condensed myelin sheaths (white arrows) were observed in a Taxotere™-treated group. (F) DTX-induced edema (n=5). Accumulative injections of Taxotere™ induced paw edema. However, no changes were observed in the in Nufs-DTX-treated group (***P<0.001, Student’s t-test between the Taxotere™ group and the control group).
Abbreviations: ALN, alanine transaminase; AST, aspartate transaminase; BUN, blood urea nitrogen; CBC, complete blood count; DTX, docetaxel; Nufs, nanoparticulation using fat and supercritical fluid; WBCs, white blood cells.