Nose-to-brain delivery of temozolomide-loaded PLGA nanoparticles functionalized with anti-EPHA3 for glioblastoma targeting

Figures & data

Table 1. Single factor study of TBE-loaded P-NPs obtained under different conditions.

Sample	Different conditions	Diameter (nm)		Drug loading (%)
Oil-phase composition (1mL)
1	dichloromethane	200.5		2.85
2	dichloromethane:acetone = 1:1	147.8		1.65
3	dichloromethane:acetone = 4:1	121.6		3.26
Oil/water ratio
4	1:3	131.1		2.92
5	1:5	142.2		2.39
6	1:10	190.3		0.89
Polymer concentration (mg/mL)
7	5	144.5		1.68
8	10	145.7		2.34
9	20	150.8		1.17
Theoretical drug loading (%)
10	5	144.9		0.78
11	10	141.8		2.49
12	15	144.2		2.10

Table 2. Characterization of TBE-loaded NPs.

Formulation	Diameter (nm)	Polydispersity Index	Zeta potential (mV)	Drug loading (%)
P-NPs	125.1 ± 3.5	0.084 ± 0.020	−21.05 ± 1.5	3.54 ± 0.51
T/P-NPs	130.3 ± 4.5	0.102 ± 0.042	+25.18 ± 2.3	3.23 ± 0.43
anti-EPHA3-T/P-NPs	145.9 ± 8.7	0.121 ± 0.035	+23.08 ± 2.5	3.02 ± 0.68

NPs: nanoparticles.

Figure 1. In vitro TBE release profiles of free TBE, P-TBE-NPs, T/P-TBE-NPs, and anti-EPHA3-T/P-TBE-NPs.

Figure 2. (A) Fluorescence microscopy images of C6 cells incubated with Nile red-loaded NPs. (B) Mean percentages of coumarin-6 NP uptake by C6 cells, as determined by flow cytometry. Values represent the mean ± SD (n = 3). **p < .01 versus P-NPs; ##p < .01 versus T/P-NPs. NPs: nanoparticles.

Figure 3. In vivo and brain distribution of DiR- and coumarin-6-loaded NPs in glioma-bearing rats. (A) In vivo fluorescence imaging at predetermined time points after intranasal administration of DiR-loaded NPs. (B) In vivo and excised tissues imaging of anti-EPHA3-modified DiR-loaded NPs at 4 h after intranasal and intravenous administration. (C) Fluorescence microscopy images of the brain, acquired 4 h after intranasal administration of coumarin-6-loaded NPs to glioma-bearing rats. Green: coumarin-6; blue: Hoechst 33342 (nuclei); yellow arrows point to the tumor site. DiR, 1,-1-dioctadecyl-3,-3,-3′,-3′-tetramethylindotricarbocyanine; NPs: nanoparticles.

Figure 4. Effects of treatments with TBE-loaded NPs on glioma-bearing rats. (A) Survival curves of glioma-bearing rats after different treatments (n = 10 each). (B) Apoptosis of glioma cells in glioma-bearing rats treated with saline, free TBE, P-TBE-NPs, T/P-TBE-NPs, and anti-EPHA3-T/P-TBE-NPs. Brown: apoptotic tumor cells; blue: tumor cells; red arrows point to apoptotic cells. (C) Quantification of apoptosis in rats treated with different formulations. Values represent the mean ± SD (n = 3). *p < .05 and **p < .01 versus P-TBE-NPs; #p < .05 versus T/P-TBE-NPs. NPs: nanoparticles; TBE: temozolomide butyl ester.