Lactoferrin- and RGD-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers for gliomatosis cerebri combination therapy

Figures & data

Figure 1 Synthesis route of lactoferrin-PEG-DSPE.

Abbreviations: DSPE-PEG₂₀₀₀-MAL, distearoylphosphatidylethanolamine-polyethylene glycol (MW~2 kDa)-maleimide; Lf, lactoferrin; PEG-DSPE, polyethylene glycol-b-distearoylphosphatidylethanolamine.

Figure 2 Scheme graph of L/R-T/V-NLCs.

Abbreviations: RGD, arginine–glycine–aspartic acid; L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers.

Figure 3 The chemical structure of Lf-PEG-DSPE and ¹H-NMR spectroscopy.

Abbreviations: Lf-PEG-DSPE, lactoferrin-PEG-DSPE; L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers.

Table 1 Characterization of different vectors

Formulations	Particle size (nm)	Size distribution (PDI)	Zeta potential (mV)	EE (%)		DL (%)
				TMZ	VCR	TMZ	VCR
L/R-NLCs	133.9±3.2	0.135±0.019	31.8±2.1	N/A	N/A	N/A	N/A
L/R-T-NLCs	138.3±4.3	0.152±0.025	33.1±2.6	82.9±3.6	N/A	6.37±0.6	N/A
L/R-V-NLCs	139.7±4.1	0.166±0.028	30.9±3.0	N/A	81.4±3.7	N/A	5.5±0.4
T/V-NLCs	96.3±3.1	0.121±0.016	34.3±2.5	83.4±2.9	81.6±2.7	10.1±0.7	7.9±0.8
L-T/V-NLCs	135.5±3.8	0.163±0.023	27.6±2.9	83.1±2.8	80.8±3.3	7.8±0.6	6.1±0.5
R-T/V-NLCs	113.6±3.1	0.143±0.022	40.5±3.1	84.2±3.2	81.2±3.1	8.4±0.8	6.5±0.6
L/R-T/V-NLCs	139.3±4.9	0.187±0.021	32.4±2.7	81.9±3.4	82.2±3.2	6.7±0.7	5.3±0.5

Abbreviations: DL, drug loading; EE, encapsulation efficiency; L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers; PDI, polydispersity index; TMZ, temozolomide; VCR, vincristine.

Figure 4 The stability of NLCs evaluated by measuring the NLC sizes (A), PDI (B), TMZ EE (C), and VCR EE (D) for 90 days.

Note: Data represented as mean±SD (n=3).

Abbreviations: EE, encapsulation efficiency; L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers; PDI, polydispersity index; TMZ, temozolomide; VCR, vincristine.

Figure 5 In vitro TMZ (A) and VCR (B) release profiles of all kinds of NLCs.

Note: Data represented as mean±SD (n=3).

Abbreviations: L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers; TMZ, temozolomide; VCR, vincristine.

Figure 6 Cellular uptake efficiency of the NLCs on U87 MG cells (A) and A549 MG cells (B).

Note: Data represented as mean±SD (n=3).

Abbreviation: L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers.

Figure 7 In vitro cytotoxicity of NLCs was evaluated on U87 MG cells (A) and T98G cells (B) by MTT assay and the synergistic effect by measuring CI₅₀ (C).

Note: CI₅₀ indicates the combination index when inhibitory concentration produced 50% cell death.

Abbreviations: L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers; Fa, fraction of affected cells.

Figure 8 In vivo TMZ tissue distribution of the NLCs and solutions at 10 h (A) and 72 h (B), and VCR tissue distribution of the NLCs and solutions at 10 h (C) and 72 h (D). Data represented as mean±SD (n=8).

Note: There is no data for 0.9% saline group.

Abbreviations: L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers; TMZ, temozolomide; VCR, vincristine.

Figure 9 In vivo antitumor efficiency of NLCs evaluated using a xenograft nude mouse model.

Notes: Tumor volume (A); tumor inhibition rate (B); body weight (C). Data represented as mean±SD (n=8).

Abbreviation: L/R-T/V-NLCs, lactoferrin- and arginine–glycine–aspartic acid dual-ligand-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers.