Fig 1.
X-Ray diffraction pattern of citrate-coated iron oxide nanoparticles.
Black peaks represent citrate-coated IONps and red peaks, ICSD-data base.
Fig 2.
TEM images and size distribution of the magnetite nanoparticle (IONps).
Fig 3.
Magnetization as a function of applied field measured at room temperature (IONps).
Low field behavior is shown in the inset.
Fig 4.
Cell viability after 72-h-exposition to citrate-coated IONps and doxorubicin hydrochloride.
The human HepG2 (hepatocellular carcinoma, A) and HaCaT (spontaneously transformed keratinocytes, B) cell lines were treated with different concentrations (100 to 0.1 ug/mL) of citrate-coated IONps, and the viability was measured 72h later with MTT assay. The blue line represents the nanoparticle and the black line corresponds to doxorubicin hydrochloride (positive control).
Fig 5.
Average body weight of female Sprague-Dawley rats during the acute oral toxicity experiment.
Results were expressed as mean ± standard deviation; Groups: control (filtered water, 2 ml/animal, n = 4), citrate-coated IONps (diluted in filtered water, 2000 mg/kg, 2 ml/animal, n = 4). Statistical analysis by Student’s T test (* p < 0.05) relating to the control group.
Table 1.
Relative weight (% relative to final body weight) of heart, kidney, spleen, liver, lung and brain at day 14 of the acute oral toxicity experiment with citrate-coated IONps.
Fig 6.
Evaluation of (A) Creatinine and (B) ALT plasma levels at the 14-day of the acute oral toxicity experiment of citrate-coated IONps. Results were expressed as mean ± standard deviation; Animals: female Sprague-Dawley rats (6 to 8 weeks, weighing between 200 ± 60 g); Groups: Pre (animals before citrate-coated IONps treatment, n = 4), Post (animals at the 14-day after citrate-coated IONps treatment, n = 4). Statistical analysis by Student’s T test (* p < 0.05) relating to the basal evaluation.
Table 2.
Biochemical plasma parameters at days 0 (baseline/pre-treatment) and 14 (post-treatment) of the acute oral toxicity protocols using citrate-coated IONps.
Fig 7.
Photomicrograph of spleen section of H&E staining of Sprague-Dawley.
(A, B) spleen, (C, D) liver, (E, F) brain, (G, H) heart, (I,J) lung and (K, L) kidney. Animals: female Sprague-Dawley rats (6 to 8 weeks, weighing between 200 ± 60 g); A, C, E, G, I and K: control group (filtered water, 2 ml/animal, n = 4); B, D, F, H, J and L: citrate-coated IONps group (diluted in filtered water, 2000 mg/kg, 2 ml/animal, n = 4). Scale bar: 50μm, 10x magnification.
Fig 8.
Fe concentration analysis by Graphite Furnace Atomic Spectrometry in liver at 14-day of the acute oral toxicity experiment of the citrate-coated IONps.
Fig 9.
Fe concentration analysis by Prussian blue staining in spleen (A, C) and liver (B, D) at 14-day of the acute oral toxicity experiment of the citrate-coated IONps. Animals: female Sprague-Dawley rats (6 to 8 weeks, weighing between 200 ± 60 g); A and B: control group (potable water, 2 ml/animal, n = 4); C and D: citrate-coated IONps group (diluted in potable water, 2000 mg/kg, 2 ml/animal, n = 4). Scale bar: 50μm, 20x magnification. Note that Fe stained by Prussian blue in liver (circle and arrow) is lower concentration then spleen.
Fig 10.
Number of Prussian blue positive profiles after exposure to citrate-coated IONps.
The graphs show quantification of the average number of Prussian blue positive staining per section from Sprague-Dawley administered control (vehicle) and treated (citrate-coated IONps). A: Liver; B: Spleen. Statistical analysis by Student’s T test (* p < 0.05) relating to the control group.
Table 3.
Iron concentration analysis by Graphite Furnace Atomic Spectrometry in vital organs at day 14 of the acute oral toxicity experiment with citrate-coated IONps.