Tuftsin-Bearing Liposomes Co-Encapsulated with Doxorubicin and Curcumin Efficiently Inhibit EAC Tumor Growth in Mice

Figures & data

Figure 1 The schematic representation of the assembly of tuftsin-bearing liposomes encapsulated with DOX and CUR.

Abbreviations: DSPE-PEG, distearoyl-L-A-phosphatidylethanolamine; DOX, doxorubicin; CUR, curcumin; LPs, liposomes.

Table 1 The Size, Polydispersity Index and Entrapment Efficiency of Liposomes

Liposomes	Average Size (nm)	Polydispersity Index (PDI)	Entrapment Efficiency (%)
			CUR	DOX	Tuftsin
DOX LPS	116	0.49		97.4±0.6
CUR LPs	136	0.31	74.9±4.3
DOX-CUR LPs	132	0.27	73.2±1.5	98.1±0.3
P.Tuft LPs	125	0.35			53%
P.Tuft-CUR LPs	128	0.4	76.8±4.2		57.2%
P.Tuft-DOX LPs	111	0.35		98.4±1.4	56.9%
P.Tuft-DOX-CUR LPs	134	0.22	73.2±4.8	96.5±2.8	52.3%

Figure 2 Size determination of LPs.

Notes: Particle size distribution of Liposomes including, DOX LPs, CUR LPs, CUR-DOX LPs, P.Tuft-CUR LPs, P.Tuft-DOX LPs, P.Tuft-DOX-CUR LPs as measured by DLS.

Abbreviations: DLS, dynamic light scattering; P.Tuft, palmitoyl tuftsin; DOX, doxorubicin; CUR, curcumin.

Figure 3 Characterisation of LPs.

Notes: UV-spectrophotometric detection and quantification of entrapped DOX and CUR in different kinds of LPs. (A) Quantification of tuftsin by RP-HPLC method, tuftsin fraction eluting at 13.9 min at 227nm (B).

Abbreviation: RP-HPLC, reverse phase liquid chromatography.

Figure 4 Characterization of structural properties by Fourier transform infrared spectroscopy (FTIR).

Note: FTIR analysis of different kinds of LPs including CUR LPs, DOX LPs, P.Tuft LPs, P.Tuft-DOX LPs, P.Tuft-DOX-CUR LPs.

Figure 5 In vitro release profile of DOX and CUR.

Notes: Percent cumulative release of DOX (A); percent cumulative release of CUR (B). The experiment was carried out using dialysis membrane diffusion technique. The data are representative of three experiments. Each value represents mean ±SD of three experiments.

Figure 6 In vitro cellular uptake study.

Notes: Confocal images of HeLa cells after 12 h of incubation with different kinds of LPs at the same concentration 0.1 mg/mL; the nuclei were stained by DAPI (blue), and all of the LPs are labeled by rhodamine B (red).

Figure 7 The in vivo effects different kinds of LPs in EAC tumor-bearing mice.

Notes: Growth curve of EAC tumors in each group (A) Images of tumors removed from mice in each group after treatment (B) Individual body weight of mice after treatment (C) Individual tumor weight of mice after 30 days of treatment (D). Each value represents mean ±SD from fifteen mice *P<0.05, **P<0.01, ***P<0.001.

Figure 8 Apoptosis protein expression in tumor tissues.

Notes: Western blot analysis and quantification of Apoptosis-related proteins (A). The cell lysates were subjected to SDS-PAGE and blotted with Bax (B), p53 (C) and BCL-2 (D) antibodies. The data are representative of three experiments. Each value represents mean ±SD of three experiments.(Figure 9) *P<0.05, **P<0.01.

Figure 9 Schematic representation of p53-mediated apoptotic pathway.

Abbreviations: Bax, Bcl-2 Associated X-protein; BCL-2, B-cell lymphoma 2; p53, tumor protein p53.

Figure 10 Assessment of liver toxicity in mice on treatment with different kinds of LPs.

Notes: Quantification of plasma concentration of AST (A) and ALT (B) after 30 days of treatment with different kinds of LPs. Each value represents mean ±SD of three experiments. **P<0.01, compared with the untreated control.

Abbreviations: AST, aspartate aminotransferase; ALT, alanine aminotransferase.

Figure 11 The micrographs of H&E-stained sections of the main organs and tumors after treatment with different kinds of LPs.

Notes: Hematoxylin and eosin-stained tumor, liver, spleen and kidney tissue after treatment of mice (treatment every second day for 30 days). Angiogenesis (black arrow), hyperplasia (red arrow). The images were observed at 20× magnification.