Table 1.
In vitro studies of adverse effects of MONPs on mammalian male germ cells. The conditions where the main findings were observed are indicated in brackets.
| MONPs | Characteristics | Concentration and Exposure Time | Cell Type | Parameters | Main Findings | Reference |
|---|---|---|---|---|---|---|
| Cerium oxide |
Formula: CeO2 Size: ~7 nm SA: 400 m2/g Shape: Ellipsoidal crystallites |
0.01, 0.1, 1, 10 µg/mL 1 h |
Spermatozoa (Human) | - Sperm vitality; - DNA damage; - Uptake of NPs |
- Sperm viability higher than the normality threshold—58% - Increased DNA damage (≥0.01 µg/mL); - Accumulation of NPs at the plasma membrane, particularly along the flagellum, without internalization |
[108] |
| Iron oxide |
Formula: Fe3O4 Size: 40 nm Shape: spherical |
0.192 mg/mL 30, 45, and 60 min |
Spermatozoa (Boar) | - Motility and kinetics | - No effects on sperm motility | [109] |
| Manganese oxide |
Formula: Mn3O4 Size: ~ 20 ± 4.1 nm Shape: irregular sphere-like morphology |
0, 5, 10, 20 µg/mL 6 and 24 h |
Sertoli Cells (Rats) | - ROS production; - MMP and apoptosis; |
- Increase in ROS (5 µg/mL, 24 h); - Alterations in the mitochondrial membrane integrity and increase in the apoptotic rates (≥5 µg/mL, 24 h) |
[110] |
| Titanium oxide |
Formula: TiO2 Size: ~30–90 nm Zeta potential: −27.3 mV |
1, 10, 100 µg/mL 0, 3, 6 h |
Spermatozoa (Bufallo) | - Viability; - Acrosomal and plasma membrane integrity; - Capacitation; - Acrosome-reaction; - DNA fragmentation; - Uptake of NPs |
- Viability decrease (100 µg/mL, 3 and 6 h); - Decrease in the integrity of the plasma membrane (≥1 µg/mL, 6 h) and acrosomal membrane (100 µg/mL, 6 h); - Increase in capacitation (≥10 µg/mL, 6 h); - Increase in acrosomal reaction (≥1 µg/mL, 3 and 6 h); - Increased DNA fragmentation (≥10 µg/mL, 6 h); - Uptake of NPs mainly in the plasma membrane and sperms’ head |
[111] |
|
Formula: TiO2 Size: ~21 nm Shape: spherical Zeta potential: −124.55 ± 13.20 mV HS: 115.2 ± 11.3 nm Purity: >99.5% PDI: 0.19 |
0.1, 1, 10, 100 µg/mL 24 h |
Spermatocytes and Sertoli cells (Mouse) |
- Viability; - Apoptosis; - Uptake of NPs - Cytoskeleton; - Migration ability; - Phagocytic activity |
- Cell viability was not affected; - Increase in the early apoptosis ratio for both cells and in the late apoptosis ratio for Sertoli cells (100 µg/mL); - Dose-dependent uptake of the nanoparticles, mainly in the cytoplasm; - Disordered microtubules (spermatocytes) and microfilaments (Sertoli cells); - Decreased migration ability of spermatocytes (100 µg/mL); - Weakened phagocytic capacity of Sertoli cells (100 µg/mL) |
[112] | |
|
Formula: TiO2 Size: ~21 nm Shape: partly irregular and semispherical |
1, 10 µg/L 15, 30, 45, 90 min |
Spermatozoa (Human) | - Viability; - Motility characteristics; - DNA damage; - ROS production |
- Cell viability was not affected; - Increase in progressive and nonprogressive sperm (1, 10 µg/L for ≥ 45 min); - Increase in DNA damage (1, 10 µg/L for ≥ 30 min); - Increase in ROS production (1, 10 µg/L for ≥ 15 min) |
[113] | |
| Zinc oxide |
Formula: ZnO Size: ~50 nm Shape: amorphous |
10, 100, 500, 1000 µg/mL 45, 90, and 180 min |
Spermatozoa (Human) | - Viability | - Increase in cell death (≥100 µg/mL, 180 min and ≥ 500 µg/mL, ≥ 45 min) | [114] |
|
Formula: ZnO Size: ~70 nm Shape: spherical Dispersion: polydisperse Surface roughness: high (22.9 nm) |
0, 5, 10, 15, 20 µg/mL 3, 6, 12, and 24 h |
Leydig and Sertoli cells (Mouse) |
- Viability; - ROS production; - Uptake of NPs; - MMP and apoptosis; - DNA damage |
- Decreased viability in both cell types (≥15 µg/mL, ≥6 h); - Increase in ROS production (≥10 µg/mL, ≥6 h) - Accumulation and uptake of nanoparticles’ aggregates in the cytoplasm and nucleus; - Loss of MMP and apoptosis increase (≥15 µg/mL, 6–12 h); - DNA leakage with an increase in chromosome breaks or loss (≥15 µg/mL, ≥12 h) |
[115] | |
|
Formula: ZnO Size: 177 nm Shape: spheroid or ellipsoid Zeta potential: −27.4 ± 1.0 mV Purity: >97% |
0, 0.04, 0.08, 0.4, 0.8, 4, 8, 16 µg/mL 24 h |
Spermatocytes and Sertoli cells (Mouse) |
- Viability; - Oxidative stress indexes (ROS, GSH, MDA) of both cell types; - Membrane permeability, MMP and cytochrome c of Sertoli cells; - TNF-α and Erk1/2 levels of Sertoli cells; - Connexin-43, occludin, claudin-5, ZO-1 expression of Sertoli cells; - DNA damage of spermatocytes; - Cell cycle analysis (cyclin E2, cyclin A2, CDK2) of spermatocytes |
- Decrease in cell viability (≥8 µg/mL); - Increase in ROS and MDA levels and decrease in GSH (8 µg/mL); - Increase in membrane permeability with decrease in MMP (8 µg/mL), but no significant changes in cytochrome c (8 µg/mL); - Increase in TNF-α and phosphorylation of Erk1/2 (8 µg/mL);- Decrease in claudin-5, occludin, ZO-1 and connexin-43 expression (8 µg/mL); - Increase in p-Chk1, p-Chk2 and ϒ-H2AX expression but decrease in APE1 (8 µg/mL) but DNA damage can be partly rescued by antioxidants; - Increase in cyclin E2, cyclin A2, CDK2 expression with an increase of cell numbers in the S phase (8 µg/mL) |
[116] | |
|
Formula: ZnO Size: 20–40 nm Shape: spherical HS: 75 nm |
0–200 µg/mL 1, 4, and 12 h |
Leydig cells (Mouse) | - Viability; - Cell morphology; - Uptake of NPs; - Apoptosis; - Oxidative stress indexes (SOD, CAT); - Steroidogenesis-related genes expression (StAR, P450scc); - Antioxidant enzyme related gene (SOD); - Testosterone levels in cells’ supernatant |
- Decrease in cell viability (≥2 µg/mL, ≥1 h); - Loss of normal morphology (≥5 µg/mL, 4 h); - Randomly dispersed agglomerates of NPs in the cytoplasm, autophagosomes, autolysosomes, mitochondria and in nuclear membranes (50 µg/mL, 4 h); - Apoptosis increase (5 or 20 µg/mL, 4 h); - Increase in SOD (1, 5 µg/mL, 4 h and 5, 20, 50 µg/mL, 12 h), CAT (1, 5, 20 µg/mL, 4 h and 5, 20 µg/mL, 12 h) activity; - Increase in StAR (1, 5 µg/mL, 4 h and 1 µg/mL, 12 h) and P450scc expression (1, 5 µg/mL, 4 h); - Decrease in SOD mRNA (1, 5 µg/mL, 4 h); - Increase in testosterone production (2 µg/mL, 12 h) |
[117] | |
|
Formula: ZnO Size: 30 nm Zeta potential: 38.25 ± 1.06 mV HS: 66.36 ± 0.93 nm |
0, 2, 3, 4, 8 µg/mL 24 h |
Leydig cells (Mouse) |
- Viability; - Oxidative stress indexes (GPx, GSH, SOD, MDA); - Apoptosis-related proteins (cleaved Casp-8 and Casp-3, Bcl-2, Bax); - Autophagy-related proteins (Atg-5, Beclin-1) and LC3-II/LC3-I ratio |
- Decrease in cell viability (≥3 µg/mL); - Increase in MDA levels (≥3 µg/mL) and decrease in SOD, GSH (≥3 µg/mL) and GPx (≥2 µg/mL) levels; - Increase in the expression of cleaved Casp-8, Casp-3 and Bax and decrease in Bcl-2 expression; - Increase in LC3-II to LC3-I ratio and Atg-5 and Beclin-1 expression (4 µg/mL) |
[118] | |
|
Formula: ZnO Size: 88 nm SA: 12 m2/g Shape: spherical Crystal structure: hexagonal wurtzite Zeta potential: −15 mV (pH = 6) and −55 mV (pH = 12) |
1, 5, 8, 10, 20 µg/mL 6 and 12 h |
Spermatogonia(Mouse) | - Viability; - Apoptosis and necrosis; - ROS production; - DNA damage; - Cytoskeleton dynamics; - Nucleoskeleton dynamics; - Nuclei morphological changes |
- Decrease in cell viability (20 µg/mL, 12 h); - Cell death by necrosis (20 µg/mL, 12 h); - Increase in ROS levels (20 µg/mL, 6 h and ≥5 µg/mL, 12 h); - Increase in DNA damage (20 µg/mL, ≥6 h); - Interference with microtubule and microfilament protein levels (20 µg/mL for 6 h and 12 h); - Alterations of the basal levels and distribution of the nuclear lamina and nuclear envelope proteins (20 µg/mL, 12 h); - Visible morphological deformities in the cells’ nuclei. |
[92] |
Abbreviations: Atg-5, Autophagy Related 5; Bax, Bcl2-associated X protein; Bcl-2, B cell lymphoma-2; Casp-, Caspase; CAT, Catalase; CDK2, Cyclin Dependent Kinase 2; DNA, Deoxyribonucleic Acid; Erk1/2, Extracellular Signal-Regulated Kinase 1/2; GPx, Glutathione Peroxidase; GSH, Reduced Glutathione; HS, Hydrodynamic Size; MDA, Malondialdehyde; MMP, Mitochondrial Membrane Potential; PDI, Polydispersity Index; P450scc, Cytochrome P450 side-chain cleavage enzyme; ROS, Reactive Oxygen Species; SA, Surface Area; SOD, Superoxide Dismutase; StAR, Steroidogenic Acute Regulatory Protein; TNF-α, Tumor Necrosis Factor Alpha; ZO-1, Zonula Occludens-1.