Estimation of silver nanoparticles effect on the reproductive health of female Wistar rats

Figures & data

Figure 1. Illustrate the XRD pattern of silver nanoparticles. GT

Figure 2. Illustrate the spherical shape of silver nanoparticles with homogenous in shape and size without any agglomeration of silver nanoparticles.

Table 1. Effect of silver nano-particles on weight.

Parameters	BWC(g)	ARO(g)	RRO(%)	ALO(g)	RLO(%)	AU(g)	RU(%)
Groups
Control	17.57 ± 1.84	0.05 ± 0.00	0.02 ± 0.00	0.05 ± 0.00	0.02 ± 0.00	0.44 ± 0.07	0.21 ± 0.03
Treated 1	15.42 ± 4.77	0.13 ± 0.045	0.07 ± 0.02	0.11 ± 0.03^a	0.06 ± 0.01^a	0.35 ± 0.072	0.208 ± 0.04
Treated 2	18.0 ± 5.83	0.05 ± 0.009	0.03 ± 0.005	0.06 ± 0.009	0.04 ± 0.009	0.61 ± 0.102	0.44 ± 0.08^a, b
F-value	0.09	2.88	2.88	2.49	3.68	2.40	5.07
P-value	0.91	0.08	0.08	0.11	0.03	0.11	0.01

The data expressed as mean± standard error of the mean (n = 7)

BWC = body weight change, ARO = absolute right ovary, RRO = relative right ovary, ALO = absolute left ovary, RLO = relative left ovary, AU = absolute uterus and RU = relative uterus.

Table 2. Effect of silver nano-particle on hormones.

Parameters	Estrogenpg/ml	Progesteronepg/ml	FSHIU/L	LHIU/L
Groups
Control	37.02 ± 2.89	13.62 ± 0.169	6.47 ± 0.36	6.47 ± 0.36
Treated 1	10.25 ± 3.77^a	8.82 ± 3.08	2.22 ± 0.085^a,b	1.90 ± 0.04^a,b
Treated 2	9.50 ± 7.16^a	6.62 ± 1.32	11.25 ± 0.63^a,b	7.47 ± 0.34^b
F-value	27.28	3.40	112.4	103.4
P-value	0.00	0.079	0.00	0.00

The data expressed as mean± standard error of the mean(n = 6) . a = P < 0.05 as compared with control. b = P < 0.05 between treated groups.

Table 3. Effect of silver nano-particles on oxidative stress markers in ovarian tissue.

Parameters	MDAnmol/ml	TACmM/L
Groups
Control	28.09 ± 4.25	0.1792 ± 0.024
Treated 1	221.59 ± 20.56^a	0.065 ± 0.011^a
Treated 2	226.55 ± 31.08^a	0.055 ± 0.011^a
F-value	27.3	16.4
P-value	0.00	0.00

The data expressed as mean± standard error of the mean(n = 6). a = P < 0.05 as compared with control. b = P < 0.05 between treated groups.

Table 4. Effect of silver nano-particles on oxidative stress markers in uterine tissue.

Parameters	MDAnmol/ml	TACmM/L
Groups
Control	75.73 ± 14.27	0.467 ± 0.124
Treated 1	252.19 ± 81.21^a	0.136 ± 0.065^a
Treated 2	283.33 ± 59.96^a	0.118 ± 0.030^a
F-value	3.6	5.60
P-value	0.049	0.019

The data expressed as mean± standard error of the mean (n = 6). a = P < 0.05 as compared with control. b = P < 0.05 between treated groups.

Figure 3. Photomicrograph of ovarian tissue of control adult female rats showing normal histology of different ovarian sections. GF = Graafian follicle, G = Granulosa, CL = Corpus Luteum, SF = Secondary Follicle, F = Follicle.

Figure 4. Photomicrograph of ovarian sections of adult female rats treated with (2 mg/kg of AgNps) low dose H&E stain. Showing: from (a to e) Degenerated tertiary follicle (black arrow), the atretic follicle (Orange arrow), the primary follicle (green arrow), high level of vacuolation with degenerated cells (bent arrow), corpus luteum (blue arrow). High percentage of apoptotic cells with pyknotic nuclei (yellow arrow), hemorrhage of blood vessels, dilated blood vessels with many RBCs (white arrow). High vacuolar degeneration in zona granulosa of the secondary follicle and stroma (black arrow), Degeneration in theca interna and cumulus oophorus of the tertiary follicle and slight degeneration in the stroma (black arrow).

Figure 5. Photomicrograph of ovarian sections of adult female rats treated with (4 mg/kg of AgNps) high dose H&E stain. Showing: from (a to e) corpus luteal (blue), the atretic follicle (Orange arrow), hemorrhage, and dilated blood vessels (green arrow), degenerated stromal cells (double head arrow), Excessive percentage of vacuolation and degeneration (bent arrow), and a high percentage of apoptotic cells with pyknotic nuclei (red arrow). Degenerated tertiary follicle containing antrum (star), degenerated follicle (black arrow).

Figure 6. Photomicrograph of uterine tissue of control adult female rats showing normal histology of different uterine sections, normal blood vessels (black arrow), and normal perimetrium layer (head arrow). MI = Myometrium I (longitudinal muscle layer), M II = Myometrium II (circular muscle layer), G = Gland, E = endometrium.

Figure 7. Photomicrographs of uterus of female rats treated with (2 mg/kg AgNps).H&E stain. P = Perimetrium, M = Myometrium, E = Endometrium.

Showing: from (A to E) uterine tissue showing degeneration in the perimetrium, endometrium, and luminal epithelium in addition to vacuolation (black arrow), congested blood vessels in perimetrium (orange arrow), enlargement in the uterine gland (blue arrow). Inflammation congested blood vessels (orange arrow), and apoptotic cell with pyknotic nuclei (red arrow) and hyperplasia of epithelial cells (yellow arrow). Also, there is an increase in number of uterine glands (blue arrow) in addition to complete degeneration (black arrow).

Figure 8. Photomicrographs of uterine tissue of female rats treated with (4 mg/kg AgNps).H&E stain. L = Lumen.

Showing: from (A to D) degeneration and apoptosis in myometrium and endometrium in addition to vacuolation (black arrow), inflammation (orange arrow), luminal epithelium showing a high level of folding, disorganization, apoptosis, and hyperplasia (yellow arrow) and lose their columnar shape. Complete vacuolar degeneration (black arrow). Apoptotic cells with pyknotic nuclei (red arrow).

Figure 9. Immunohistochemistry showing caspase-3 streptavidin-biotin- peroxidase staining method in ovarian tissue of control adult female rat showing normal expression of immune stain caspase-3.

GF = Graafian follicle, F = Follicle, S = Stroma, CL = Corpus Luteum

Figure 10. Immunohistochemistry showing caspase-3 streptavidin-biotin- peroxidase staining method in ovarian tissue of adult female rats treated with (2 mg/kg of AgNps) low dose. Showing positive and moderate expression of immune stain caspase-3.

Figure 11. Immunohistochemistry showing caspase-3 streptavidin-biotin- peroxidase staining method in ovarian tissue of adult female rat treated with (4 mg/kg of AgNps) high dose, Showing positive and extreme expression of immune stain caspase-3.

Figure 12. Immunohistochemistry showing caspase-3 streptavidin-biotin- peroxidase staining method in the uterus of control adult female rats showing normal expression of immune stain caspase-3. LM = Longitudinal Muscle, CM = Circular Muscle, E = endometrium, G = Gland.

Figure 13. Immunohistochemistry showing caspase-3 streptavidin-biotin- peroxidase staining method in the uterus of adult female rat treated with (2 mg/kg of AgNps) low dose showing positive and moderate expression of immune stain caspase-3.

Figure 14. Immunohistochemistry showing caspase-3 streptavidin-biotin- peroxidase staining method in the uterus of adult female rat treated with (4 mg/kg of AgNps) high dose Showing positive and extreme expression of immune stain caspase-3.

Table 5. Effect of silver nanoparticles on Immunohistochemistry expression for activated caspase-3.

Parameters	Ovarian follicles	Ovarian stroma	Corpora luteal	Uterus
Groups
Control	0.166 ± 0.01	0.167 ± 0.004	0.186 ± 0.006	0.125 ± 0.004
Treated 1	0.297 ± 0.023^a	0.29 ± 0.026^a	0.306 ± 0.019^a	0.311 ± 0.014^a
Treated 2	0.309 ± 0.025^a	0.28 ± 0.01^a	0.313 ± 0.025^a	0.389 ± 0.015^a
F-value	13.58	10.89	15.89	127.11
P-value	0.00	0.00	0.00	0.00

The data expressed as mean± standard error of the mean (n = 3). a = P < 0.05 as compared with control. b = P < 0.05 between treated groups.