A Novel Delivery System of RGD-HSA Loaded GEM/CUR Nanoparticles for the Treatment of Pancreatic Cancer Therapy

Figures & data

Table 1 The Characteristics of Different Formulations: Particle Size, Entrapment Ratio, Polydispersity Index and ζ-Potential. (Mean ± SD, n=3)

	Particle Size (nm)	Encapsulation Ratio (%)	Polydispersity Index (PDI)	ζ-potential (mV)
HSA-GEM/CUR NPs	113.6±6.1	81.4±5.1	0.131±0.06	−15.9±2.4
RGD-HSA-GEM/CUR NPs	115.6±5.7	82.2±4.5	0.139±0.03	−17.3±1.6

Figure 1 Transmission electron microscope of HSA-GEM/CUR NPs (A); RGD-HSA-GEM/CUR NPs (B); RGD-HSA-GEM/CUR NPs after 6 months (C).

Table 2 Stability Test Observations of the RGD-HSA-GEM/CUR NPs at Room Temperature (Mean ± SD, n=3)

	Time (m)	Physical Changes	Particle Size (nm)	Encapsulation Ratio (%)	Polydispersity Index (PDI)	ζ-potential (mV)
Room temperature	0	-	115.6±5.7	82.2±4.5	0.139±0.03	−17.3±1.6
	1	-	117.3±4.6	81.1±3.7	0.142±0.05	−18.5±1.8
	3	-	118.1±3.9	81.4±2.9	0.147±0.06	−17.9±1.3
	6	-	120.6±6.1	80.2±3.1	0.141±0.04	−18.1±1.4

Note: No physical change.

Figure 2 The release profile of drug in NPs in PBS (containing 0.1% Tween 80, pH 7.4) (n=6).

Figure 3 Confocal images of cellular uptake of Blank NPs (A); free GEM/CUR (B), HSA-GEM/CUR NPs (C) and RGD-HSA-GEM/CUR NPs (D) by SW1990 cells. Incubation time was 2 hours. (E) was the quantitative results. Bar=50 µm. (^ap <0.05, RGD-HSA-GEM/CUR NPs vs Blank NPs; ^bp <0.05, RGD-HSA-GEM/CUR NPs vs free GEM/CUR, ^cp <0.05, RGD-HSA-GEM/CUR NPs vs HSA-GEM/CUR NPs).

Table 3 The IC50 Values of Free Gemcitabine/Curcumin (GEM/CUR), Human Serum Albumin-Gemcitabine/Curcumin Nanoparticles (HSA-GEM/CUR NPs) and Arginine Glycine Peptide-Human Serum Albumin-Gemcitabine/Curcumin Nanoparticles (RGD-HSA-GEM/CUR NPs) Treated SW1990 Cells (Mean ± SD, n=6)

Formulations	SW1990 IC50 (µM)
Free GEM/CUR	51.2±3.24*
HSA-GEM/CUR NPs	18.3±2.33*
RGD-HSA-GEM/CUR NPs	11.1±1.13

Note: *p<0.05 vs the group of RGD-HSA-GEM/CUR NPs.

Figure 4 In vitro viability of different NPs formulations in SW1990 cells. Data represents mean ± SD (n = 3). A Cell viability cultured with Blank NPs; free GEM/CUR, HSA-GEM/CUR NPs and RGD-HSA-GEM/CUR NPs at various concentrations of GEM/CUR after 24 h. (^ap <0.05, RGD-HSA-GEM/CUR NPs vs Blank NPs; ^bp <0.05, RGD-HSA-GEM/CUR NPs vs free GEM/CUR, ^cp <0.05, RGD-HSA-GEM/CUR NPs vs HSA-GEM/CUR NPs).

Table 4 The Mean AUC_0-t and Target Index of Different Formulations of NPs in Different Organs of Mice (Mean ± SD, n=8)

AUC0-t (ngh/g)	GEM(Free GEM/CUR)	GEM(HSA-GEM/CUR NPs)	GEM(RGD-HSA-GEM/CUR NPs)	CUR(Free GEM/CUR)	CUR(HSA-GEM/CUR NPs)	CUR(RGD-HSA-GEM/CUR NPs)
Tumor	4096.5±327.2	7096.8±540.9	9637.0±656.3	6055.0±398.5	6617.0±412.3	8832.8±433.6
Heart	5364.8±298.3	4188.3±329.7	3791.3±327.5	5379.8±376.3	5662.5±309.2	4032.8±309.2
Liver	5201.5±343.2	2871.6±293.4	2653.9±253.2	5629.4±329.1	5410.7±314.3	2432.6±298.3
Spleen	5688.8±432.1	4845.5±378.5	5239.5±367.5	6340.8±436.4	5775.3±331.3	5172.3±287.5
Lung	5901.8±309.2	5445.5±438.6	5739.0±219.4	7564.5±459.4	6376.5±378.4	6583.0±423.5
Kidney	5624.0±312.5	6141.3±548.7	3487.5±328.1	4995.5±321.4	5305.5±298.7	2673.5±278.3
Plasma	4351.3±298.3	5609.0±438.6	3741.3±287.5	4029.0±309.2	5279.0±310.2	4901.3±309.2
TI(Tumor/Heart)	0.76	1.69	2.54^ab	1.13	1.17	2.19^cd
TI(Tumor/Liver)	0.79	2.47	3.63^ab	1.08	1.22	3.63^cd
TI(Tumor/Spleen)	0.72	1.46	1.84^a	0.95	1.15	1.71^cd
TI(Tumor/Lung)	0.69	1.30	1.68^a	0.80	1.04	1.34^c
TI(Tumor/Kidney)	0.73	1.16	2.76^ab	1.21	1.25	3.30^cd
TI(Tumor/Plasma)	0.94	1.27	2.58^ab	1.50	1.25	1.80^d

Notes: ^ap<0.05: GEM(RGD-HSA-GEM/CUR NPs) vs GEM(free GEM/CUR); ^bp<0.05: GEM(RGD-HSA-GEM/CUR NPs) vs GEM(HSA-GEM/CUR NPs); ^cp<0.05: CUR(RGD-HSA-GEM/CUR NPs) vs CUR(free GEM/CUR); ^dp<0.05: CUR(RGD-HSA-GEM/CUR NPs) vs CUR(HSA-GEM/CUR NPs).

Figure 5 In vivo biodistribution study of modified and non-modified GEM/CUR NPs (n=8). (A) tumor, (B) heart, (C) liver, (D) spleen, (E) lung, (F)kidney and (G) plasma. (^ap <0.05, RGD-HSA-GEM/CUR NPs vs free GEM/CUR, ^bp <0.05, RGD-HSA-GEM/CUR NPs vs HSA-GEM/CUR NPs).

Figure 6 The in vivo imaging of DiR-loaded Blank NPs; free GEM/CUR, HSA-GEM/CUR NPs and RGD-HSA-GEM/CUR NPs in tumor bearing nude mice at 12 h (n=3).

Figure 7 (A) The growth curves of SW1990 tumors in mice. (B) The changes in the body weight of SW1990 tumor-bearing mice. (^ap <0.05, RGD-HSA-GEM/CUR NPs vs Blank NPs; ^bp <0.05, RGD-HSA-GEM/CUR NPs vs free GEM/CUR, ^cp <0.05, RGD-HSA-GEM/CUR NPs vs HSA-GEM/CUR NPs).

Figure 8 Analysis of apoptotic pathway proteins. Western Blot result of apoptotic protein expression (A) and expression of actin (B) on SW1990 cells after incubated with free GEM/CUR, HSA-GEM/CUR NPs and RGD-HSA-GEM/CUR NPs for 24 h (GEM/CUR 0.5 μM). (^ap <0.05, RGD-HSA-GEM/CUR NPs vs Control; ^bp <0.05, RGD-HSA-GEM/CUR NPs vs free GEM/CUR).