Table 3 Summary information of the biological endpoints measured in the reproductive system toxicology

Cell viability

MTT, MTS, LDH, CCK-8 assay

[96, 97, 105, 107, 112]

Oxidative stress

ROS Production

H2DCFDA

[95, 97, 105]

Lipid peroxidation,

MDA content

In testicular tissue and cells

ELISA, Western Blot, qRT-PCR

[89, 90, 94, 96, 98, 109]

Enzymatic/non-enzymatic antioxidants

GPX, CAT, SOD, TBARS, TAOC, GSH

in Sperm, Seminal Plasma, and Blood, serum, testicular tissue homogenate and in vitro cells

ELISA, Western Blot, qRT-PCR

[46, 88, 89, 96, 104, 155]

DNA damage

DNA Strand breaks,

In cells and the testis tissue

Comet Assay, DNA microarray analysis,

DNA fragmentation

[46, 88, 89, 96, 107, 109]

Mitochondrial damage

Cell metabolic activity

MTT assay

[112]

Mitochondrial membrane potential

Quantitative method uses a tetraphenylphosphonium ion (TPP+)-sensitive electrode, or by fluorimetric methods

[97]

Apoptosis, in germ Sertoli and Leydig cell

P53, BAX, bcl-2 gene expressions or cell apoptosis

Western Blot, qPCR, TUNEL Assay Flow Cytometry

[90, 93, 97, 112, 113, 166]

Reproductive hormone levels

Hormonal Assessment

Serum Testosterone, LH and FSH,

P450scc, StAR, Hsd3b, and Cyp17a1 gene expression

Radioimmunoassay [167], ELISA, qRT-PCR, chemiluminescent protein immunoassay

[46, 89, 91, 98, 100, 132, 174]

Sperm evaluations

Morphology, Motility, Concentration, Count, Viability, Plasma membrane intergrity

Eosin/nigrosine staining, hemocytometer, Microscope

[46, 92, 95, 103, 146, 166, 175]

Acrosome status

Fluorescence assessment (eg.chlortetracycline fluorescence assay)

[65, 146, 175]

Sperm DNA integrity

Toluidine blue staining, Aniline blue staining, Acridine orange staining Eosin–nigrosine-staining

[38, 90, 103, 123, 148]

Sperm DNA damage

Comet assay

[109, 148]

The mitochondrial activity

Activity of cytochrome c oxidase

[65, 108]

Spermatogenesis

The transcript expression of Gnrh1, Ar, Cyp11a1, Hsd3b1, Hsd17b3, Srd5a1, Cyp19a1, Star by qPCR

Intratesticular steroid metabolism enzyme protein level aromatase (Aro) and 5α-reductase type 1 (Srd5a1) measurements

[108, 132]

Determination of silver concentration in organs

ICPMS, UV/vis proton spectrophotometery

[38, 114, 176]

Histopathological Examination

The seminiferous tubules area, circumference, and diameter,

Testis index and histological structure of testicular tissues

Hematoxylin and Eosin light microscope Transmission electron microscopy (TEM)

[46, 100, 103, 123]