T₁–T₂ molecular magnetic resonance imaging of renal carcinoma cells based on nano-contrast agents

Figures & data

Table S1 T₁ and T₂ relaxation time of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex prepared with different concentrations of Fe₃O₄ nanoparticles $(\overline{\mathrm{X}}\pm \mathrm{S})$ (n=4)

Group	Fe3O4 concentration (mg)	T1 relaxation time (ms)	T2 relaxation time (ms)
1	1 mg	318.25±3.86	66.25±2.22
2	3 mg	448.75±7.93	65.25±2.50
3	5 mg	453.25±6.29	65.00±2.58

Table S2 T₁ and T₂ relaxation time of the mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex, Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex and Fe₃O₄@mSiO₂ nanocomplex $(\overline{\mathrm{X}}\pm \mathrm{S})$ (n=4)

Group	T1 relaxation time (ms)	T2 relaxation time (ms)
mSiO2/PDDA/Gd2O3	1,293.20±63.13*	557.00±15.58a
Fe3O4@mSiO2/PDDA/Gd2O3	846.75±26.50#	146.50±26.25
Fe3O4@mSiO2	219.50±5.80	149.25±12.63

Notes:

* Refers to statistically significant differences when compared with the other two groups using one-way ANOVA (p<0.05).

# Refers to statistically significant differences when compared with the other two groups using one-way ANOVA (p<0.05).

a Refers to statistically significant differences when compared with the other two groups using one-way ANOVA (p<0.05).

Table S3 T₁ relaxation time of NIH-3T3 mouse fibroblast cells and 786-0 renal carcinoma cells treated with 1 mg mL⁻¹ Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (−) and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe (+) $(\overline{\mathrm{X}}\pm \mathrm{S})$ (n=4)

Group	NIH-3T3 cells	786-0 cells
Control	1,823.80±18.84	1,752.80±2.50
AS1411(+)	1,740.80±21.70*	902.25±2.21#,a
AS1411(−)	1,751.00±9.13*	1,241.20±3.40#,a

Notes:

* Refers to statistically significant differences when compared with control group using one-way ANOVA (p<0.05).

# Refers to statistically significant differences when compared with the control group using one-way ANOVA (p<0.05).

a Refers to statistically significant differences between NIH-3T3 and 786-0 cells with LSD Duncan (p<0.05).

Table S4 T₂ relaxation time of NIH-3T3 mouse fibroblast cells and 786-0 renal carcinoma cells treated with 1 mg mL⁻¹ Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (−) and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe (+) $(\overline{\mathrm{X}}\pm \mathrm{S})$ (n=4)

Group	NIH-3T3 cells	786-0 cells
Control	132.50±9.95	129.00±2.94
AS1411(+)	67.00±2.16*	41.25±2.63#,a
AS1411(−)	65.50±2.65*	53.75±2.75#,a

Notes:

* Refers to statistically significant differences when compared with the control group using one-way ANOVA (p<0.05).

# Refers to statistically significant differences when compared with the control group using one-way ANOVA (p<0.05).

a Refers to statistically significant differences NIH-3T3 and 786-0 cells with LSD Duncan (p<0.05).

Figure 1 Characterization of the assembly process of Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ with zeta potential (A), hydrodynamic diameter determination (B), and the fabrication of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe with fourier transform infrared absorption (FTIR) spectra (C) and UV-vis spectra (D).

Figure 2 T₁-weighted magnetic resonance (MR) images and T₁-map images as well as T₂-weighted MR images and T₂-map images of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex prepared with different concentrations of BSA-Gd₂O₃ NPs. 3.0 T human magnetic resonance scanner was used.

Figure 3 T₁ and T₂ relaxation time of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex prepared with different concentrations of BSA-Gd₂O₃ NPs. BSA-Gd₂O₃ NPs (0 µM) were treated as the control group, and other groups were all compared with the control group with one-way ANOVA. *Refers to statistically significant differences compared with the control group with one-way ANOVA (p<0.05). ^#Refers to statistically significant differences compared with control group with one-way ANOVA (p<0.05). A 3.0 T human magnetic resonance scanner was used.

Figure 4 T₁ relaxivity curves (A) and T₁-weighted magnetic resonance (MR) images (B) of Gd-DTPA (1), BSA-Gd₂O₃ NPs (2), and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (3) with various Gd concentrations. A 3.0 T human MR scanner was used.

Figure 5 T₂ relaxivity curves and T₂-weighted magnetic resonance (MR) images of the Fe₃O₄ (A), and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (B) with various Fe₃O₄ nanoparticle concentrations. A 3.0 T human MR scanner was used.

Figure 6 Cell viability of 786-0 renal carcinoma cells and NIH-3T3 mouse fibroblast cells after exposure to various concentrations of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex, determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

Figure 7 Iron and gadolinium ions released from the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex in cell culture medium over time (A) and in different pH solutions incubated for 24 h (B).

Figure 8 T₁-weighted magnetic resonance (MR) images and T₁-map images (A) as well as T₂-weighted MR images and T₂-map images (B) of 786-0 renal carcinoma cells treated with different concentrations of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (−) and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe (+) (0, 0.5, 1, 2.5 mg/mL⁻¹). A 3.0 T human MR scanner was used.

Figure 9 T₁ (A) and T₂ relaxation time (B) of 786-0 renal carcinoma cells treated with different concentrations of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (−) and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe (+).

Notes: *Refers to statistically significant differences compared with the control group using one-way ANOVA (p<0.05). ^#Refers to statistically significant differences compared with the control group with one-way ANOVA (p<0.05). ^aRefers to statistically significant differences within group with LSD Duncan (p<0.05). ^bRefers to statistically significant differences within group with LSD Duncan (p<0.05). ^cRefers to statistically significant differences within group with LSD Duncan (p<0.05). ^dRefers to statistically significant differences within group with LSD Duncan (p<0.05). A 3.0 T human magnetic resonance scanner was used.

Figure 10 The concentrations of Gd in the heart, lung, liver, and kidney at 1 h and 24 h post-intravenous injection of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex as determined by inductively coupled plasma-mass spectrometry (n=3, *p<0.05, **p<0.01).

Figure 11 Hematologic analysis of mice at 1 week post-injection of normal saline (black bar) and the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (red bar).

Notes: (A) White blood cells (WBC), (B) red blood cells (RBC), (C) platelets (PLT), (D) mean corpuscular volume (MCV), (E) hemoglobin (HGB), (F) mean corpuscular hemoglobin concentration (MCHC), (G) mean corpuscular hemoglobin (MCH), and (H) hematocrit (HCT) levels in the blood. ***Refers to statistically significant differences compared with the normal saline group (p<0.001, n=3).

Figure 12 Biochemical analysis of mice at 1 week post-injection of normal saline (black bar) and the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (red bar).

Notes: (A) Alanine transaminase (AST), (B) aspartate transaminase (ALT), (C) total protein (TP), (D) albumin (ALB), (E) total bilirubin (TBIL), (F) blood urea nitrogen (BUN), and (G) creatinine (CREA) levels in the blood; (n=3).

Figure 13 (A) T₁-weighted in vivo magnetic resonance imaging (MRI) images of mice post-injection of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex at different time points (0, 15 min, 6 h, and 24 h). The signal intensities in the kidneys (B) and bladder (C) at different time points after intravenous injection of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex. The red arrow indicates the kidney.

Scheme 1 Schematic illustration of the fabrication process of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobes.

Figure S1 Transmission electron microscopy characterizations of Fe₃O₄ nanoparticles (NPs) (A) and Fe₃O₄@mSiO₂ NPs (B). The insert in (A) is the distribution of Fe₃O₄ NP size.

Figure S2 X-ray powder diffraction patterns of the Fe₃O₄, Fe₃O₄@mSiO₂ and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex.

Figure S3 T₁-weighted magnetic resonance (MR) images and T₁-map images as well as T₂-weighted MR images and T₂-map images of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex prepared with different concentrations of Fe₃O₄ nanoparticles. A 3.0 T human MR scanner was used.

Figure S4 T₁-weighted magnetic resonance (MR) images as well as T₂-weighted MR images of the mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex, Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex, and Fe₃O₄@mSiO₂ nanocomplex. A 3.0 T human MR scanner was used.

Figure S5 T₁-weighted magnetic resonance (MR) images and T₁-map images as well as T₂-weighted MR images and a T₂-map images of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe. No significant difference of T₁ and T₂ relaxation time could be observed after the AS1411 aptamer modification. A 3.0 T human MR scanner was used.

Figure S6 T₁-weighted magnetic resonance (MR) images and T₁-map images (A) as well as T₂-weighted MR images and T₂-map images (B) of NIH-3T3 cells treated with 1 mg mL⁻¹ Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex (−) and Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃-AS1411 nanoprobe (+). A 3.0 T human MR scanner was used.

Figure S7 (A) T₂-weighted in vivo magnetic resonance imaging (MRI) images of mice post-injection of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex at different time points (0, 15 min, 6 h, and 24 h). The signal intensities in the kidneys (B) and bladder (C) at different time points after intravenous injection of the Fe₃O₄@mSiO₂/PDDA/BSA-Gd₂O₃ nanocomplex. The red arrow indicates the kidney.