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. 2023 Feb 14;24(4):3784.
doi: 10.3390/ijms24043784.

Comparative Evaluation of the Effects of Amorphous Silica Nanoparticles on the Erythrocytes of Wistar Normotensive and Spontaneously Hypertensive Rats

Zannatul Ferdous  1 , Ozaz Elzaki  1 , Sumaya Beegam  1 , Nur Elena Zaaba  1 , Saeed Tariq  2 , Ernest Adeghate  2 , Abderrahim Nemmar  1   3
Affiliations

Comparative Evaluation of the Effects of Amorphous Silica Nanoparticles on the Erythrocytes of Wistar Normotensive and Spontaneously Hypertensive Rats

Zannatul Ferdous et al. Int J Mol Sci. .

Abstract

Silica nanoparticles (SiNPs) are one of the most widely used nanomaterials. SiNPs can encounter erythrocytes and hypertension is strongly linked to abnormalities in the functional and structural characteristics of erythrocytes. As little is known about the combinatorial effect of SiNP-hypertension interactions on erythrocytes, the aim of this work was to study the effects triggered by hypertension on SiNPs induced hemolysis and the pathophysiological mechanism underlying it. We compared the interaction of amorphous 50 nm SiNPs at various concentrations (0.2, 1, 5 and 25 µg/mL) with erythrocytes of normotensive (NT) and hypertensive (HT) rats in vitro. Following incubation of the erythrocytes, SiNPs induced significant and dose-dependent increase in hemolysis. Transmission electron microscopy revealed erythrocyte deformity in addition to SiNPs taken up by erythrocytes. The erythrocyte susceptibility to lipid peroxidation was significantly increased. The concentration of reduced glutathione, and activities of superoxide dismutase, and catalase were significantly increased. SiNPs significantly increased intracellular Ca2+. Likewise, the concentration of the cellular protein annexin V and calpain activity was enhanced by SiNPs. Concerningly, all the tested parameters were significantly enhanced in erythrocytes from HT rats compared to NT rats. Our results collectively demonstrate that hypertension can potentially exacerbate the in vitro effect induced by SiNPs.

Keywords: erythrocytes; hypertension; silica nanoparticles.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Transmission electron micrographs of the erythrocytes (RBCs) of normotensive (NT) ((a–e): (a) NT-saline; (b) NT 0.2; (c) NT 1:0; (d) NT 5.0; (e) NT 25 µg/mL nanoparticles) and hypertensive (HT) ((f–j): (f) HT-saline; (g) HT 0.2; (h) HT 1:0; (i) HT 5.0; (j) HT 25 µg/mL nanoparticles) rats after treatment with silica nanoparticles (SiNPs). Abnormal RBCs (thin arrows) were observed in NT rats after the addition of nanoparticles at a concentration of 1, 5 or 25 µg/mL. In contrast, the RBCs of HT rats show abnormal shapes (empty arrows) even when treated with saline (f). The addition of nanoparticles increased the number of deformed RBCs. Scale bar = 2 µm.
Figure 2
Figure 2
Transmission electron micrographs of the erythrocytes of normotensive (NT) ((a–e): (a) NT-saline; (b) NT 0.2; (c) NT 1:0; (d) NT 5.0; (e) NT 25 µg/mL nanoparticles) and hypertensive ((f–j): (f) HT-saline; (g) HT 0.2; (h) HT 1:0; (i) HT 5.0; (j) HT 25 µg/mL nanoparticles) rats after treatment with silica nanoparticles (SiNPs). A strong interaction between RBCs and nanoparticles were observed after treatment with nanoparticles at a concentration of either 5 or 25 µg/mL. The 50 nm SiNPs entered RBCs and caused destruction and eventual sloughing (arrow) of red blood corpuscles in HT rats (i,j). The extent of RBC damage is less severe after attachment of nanoparticles (empty circles) onto RBCs of NT rats. Scale bar = 2 µm.
Figure 3
Figure 3
Numbers of deformed red blood cells (RBCs) (per power field, ×11,100) of normotensive (NT) and hypertensive HT) rats after treatment with various concentrations (µg/mL) of silica nanoparticles (SiNPs). Data are mean ± SEM (n = 5 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm-Sidak multiple comparisons test. * p < 0.05, *** p < 0.001, **** p < 0.0001 vs. respective control and Δ p < 0.05, ΔΔ p < 0.01, ΔΔΔ p < 0.001, ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 4
Figure 4
Hemolytic effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) in incubated normotensive (NT) and hypertensive (HT) rat erythrocytes. The results are expressed as % of positive control (0.1% Triton-X 100). Data are mean ± SEM (n = 8 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm–Sidak multiple comparisons test. * p < 0.05, **** p < 0.0001 vs. respective control and ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 5
Figure 5
Effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) on lipid peroxidation concentration (LPO, (a)), catalase activity (b), superoxide dismutase activity (SOD, (c)) and reduced glutathione concentration (GSH, (d)) measured in the incubation medium of erythrocytes obtained from normotensive (NT) and hypertensive (HT) rats. Data are mean± SEM (n = 8 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm-Sidak multiple comparisons test. ** p < 0.01, **** p < 0.0001 vs. respective control and ΔΔ p < 0.01, ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 6
Figure 6
Effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) on lactate dehydrogenase concentration in incubated normotensive (NT) and hypertensive (HT) rat erythrocytes. Data are mean± SEM (n = 6 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm–Sidak multiple comparisons test. **** p < 0.0001 vs. respective control and ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 7
Figure 7
Effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) on ATPase activity measured in incubated erythrocytes of normotensive (NT) and hypertensive (HT) rats. Data are mean± SEM (n = 8 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm–Sidak multiple comparisons test. * p < 0.05, **** p < 0.0001 vs. respective control and ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 8
Figure 8
Effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) on intracellular calcium concentration measured in incubated erythrocytes of normotensive (NT) and hypertensive (HT) rats. Data are mean± SEM (n = 8 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm–Sidak multiple comparisons test. ** p < 0.01, **** p < 0.0001 vs. respective control and ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 9
Figure 9
Effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) on the concentration of bound Annexin V in the incubation medium of erythrocytes of normotensive (NT) and hypertensive (HT) rats. Data are mean± SEM (n = 8 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm–Sidak multiple comparisons test. **** p < 0.0001 vs. respective control and ΔΔΔΔ p < 0.0001 vs. respective concentration in NT group.
Figure 10
Figure 10
Effect of various concentrations (µg/mL) of silica nanoparticles (SiNPs) on calpain activity measured in incubated erythrocytes of normotensive (NT) and hypertensive (HT) rats. Data are mean± SEM (n = 6 in each group). Statistical analysis by one-way analysis of variance (ANOVA) followed by the Holm–Sidak multiple comparisons test. **** p < 0.0001 vs. respective control.
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