In vitro and in vivo genotoxicity and molecular response of silver nanoparticles on different biological model systems

Figures & data

Figure 1. Cytotoxic activity of three concentrations of silver nanoparticles (μg ml^–1) from different weight ratios of silver metal (1, 3 and 5%) against MCF-7 cell line (survival %).

^**Statistically significant at p ≤ 0.01.

Figure 2. Cytotoxic activity of three concentrations of silver nanoparticles (μg ml^–1) from different weight ratios of silver metal (1, 3 and 5%) against HepG2 cell line (survival %).

Figure 3. Mitotic index of Allium cepa meristems treated with silver nanoparticles. ^aStatistically significant at p ≤ 0.01.

Figure 4. Types of chromosomal aberrations induced in Allium cepa meristems treated with silver nanoparticles. (a, b) irregular prophase; (c, d) c-metaphase; (e, f) disturbed anaphase; (g) sticky anaphase; (h) chromatin bridge; (i) chromosomal break and micronucleus in metaphase; (j) micronuclei in interphase; (k) uniucleate cell with two micronuclei (binucleate); (l) multinucleate cell in interphase.

Figure 5. Percentage of micronucleus induced in Allium cepa meristems treated with silver nanoparticles.

^aStatistically significant at p ≤ 0.05.

^bStatistically significant at p ≤ 0.01.

Figure 6. Change in protein profile of (a) Staphylococcus aureus and (b) Pseudomonas aeruginosa after treatment with silver nanoparticles.

(a) M: Marker; Sc: S. aureus control; S1%: S. aureus treated with 1% AgNPs; S3%: S. aureus treated with 3% AgNPs; S5%: S. aureus treated with 5% AgNPs.

(b) M: Marker; Pc: P. aeruginosa control; P1%: P. aeruginosa treated with 1% AgNPs;P3%: P. aeruginosa treated with 3% AgNPs; P5%: P. aeruginosa treated with 5% AgNPs.

Figure 7. DNA polymorphism based on RAPD-PCR analysis of (a) Staphylococcus aureus and (b) Pseudomonas aeruginosa treated with silver nanoparticles.

(a) M: Marker; c: S. aureus control; T1: S. aureus treated with 1% AgNPs; T2: S. aureus treated with 3% AgNPs; T3: S. aureus treated with 5% AgNPs.

(b) M: Marker; c: P. aeruginosa control; T1: P. aeruginosa treated with 1% AgNPs; T2: P. aeruginosa treated with 3% AgNPs; T3: P. aeruginosa treated with 5% AgNPs.

Figure 8. DNA polymorphism based on ISSR-PCR analysis of (a) Staphylococcus aureus and (b) Pseudomonas aeruginosa treated with silver nanoparticles.

(a) M: Marker; c: S. aureus control; T1: S. aureus treated with 1% AgNPs; T2: S. aureus treated with 3% AgNPs; T3: S. aureus treated with 5% AgNPs

(b) M: Marker; c: P. aeruginosa control; T1: P. aeruginosa treated with 1% AgNPs; T2: P. aeruginosa treated with 3% AgNPs; T3: P. aeruginosa treated with 5% AgNPs.

Table 1. Percentage of mitotic abnormalities and phase distribution of Allium cepa meristems treated with silver nanoparticles in different mitotic phases.

		Total no. of counted cells	Mitotic abnormalities	Prophase	Metaphase	Anaphase	Telophase
Control
3h	5010		5.40 ± 0.66	49.5 ± 0.75	24.1 ± 0.20	22.6 ± 0.14	3.8 ± 0.65
12h	5223		6.91 ± 0.93	54.2 ± 0.39	22.3 ± 0.81	20.4 ± 0.28	3.1 ± 1.23
1%
3 h	5149		9.17a ± 0.70	47.2 ± 0.42	22.3 ± 0.11	25.1a ± 0.61	5.4b ± 0.32
12 h	5008		10.80b ± 0.62	58.7a ± 0.10	17.5 ± 0.34	23.8a ± 0.09	0.0
3%
3 h	5114		15.03b ± 0.52	55.3a ± 0.88	20.8 ± 0.55	19.9 ± 1.01	4.0a ± 0.12
12 h	5031		20.43b ± 0.87	62.5b ± 0.37	22.6 ± 0.49	10.3 ± 0.73	4.6a ± 0.08
5%
3 h	5097		20.76b ± 1.03	60.7b ± 0.79	23.2 ± 0.27	13.5 ± 0.95	2.6 ± 0.13
12 h	5154		35.26b ± 0.41	65.2b ± 0.06	21.8 ± 0.52	11.2 ± 0.06	1.8 ± 0.29

^a Statistically significant at p ≤ 0.05.

^b Statistically significant at p ≤ 0.01.

Table 2. Percentage of different types of abnormalities induced in Allium cepa meristems after treatment with silver nanoparticles.

Concentration	Time (h)	Type of abnormality
		Irregular prophase	Stickiness	Spindle disturbance	Bridge	Break
Control	3	21	37.1	40.9	0	1
	12	18.4	33.4	48.2	0	0
1%	3	3	54	34.9	2.1	6
	12	5.7	54.6	31.1	3.2	5.4
3%	3	4.7	51.0	32.7	5.0	6.6
	12	2.1	57	35	0	0
5%	3	0	61	36.9	0	2.1
	12	1.0	59.9	36.5	1.6	1.0

Table 3a. SDS-PAGE electrophoretic profile of Staphylococcus aureus with intensities after treating with silver nanoparticles.

Molecular weight (kDa)	Marker	Control	1%	3%	5%
90		4.13	0.20	12.40	0
86		0	0	0	14.41
84		1.98	9.81	0	7.80
82		0.57	0	0	0
79		0	0	2.25	0
77		3.24	2.32	0	0
76		0	0	2.49	0
75		0	1.06	0	1.17
74		2.63	1.19	0	1.41
73		0	1.72	0.88	1.17
71		1.59	1.38	2.96	0
70		0	0	0	2.42
69		0	1.33	0	0
68		2.44	0	1.63	0
66	1	0	0	2.06	4.96
65		0	3.29	0	0
64		2.55	1.70	1.61	0
63		0	0	0.66	2.01
62		2.77	1.49	0	0
61		0	0	1.77	3.68
60		0	2.25	1.67	0
57		0	2.62	1.76	0
56		3.93	1.94	1.02	2.71
55		0	0	1.00	1.13
54		0	1.09	0	0
53		0	0	2.91	0
51		5.59	7.33	2.45	6.80
49		0	0	1.99	0
48		2.04	2.72	0.90	1.00
47		0	0	2.37	1.28
45	1	1.08	2.63	0	0
44		0	0	2.31	2.94
43		1.54	0	0	0
42		0	4.58	4.53	0
41		0	0	0	5.37
38		8.55	3.03	3.09	3.12
37		0	0	1.10	1.29
36		0.56	0	0	0
35		0	2.77	3.29	0
34		9.81	0	0	2.97
33		0	3.02	0	0
32		5.33	0	0	2.34
31		0	0	4.13	0
30		5.11	0	0	6.12
29		0	11.37	3.08	0
27		0	1.62	0	0
26		0.33	0	6.07	2.56
25	1	0	0	0	0
24		0	2.76	2.15	2.53
23		15.45	9.31	6.04	4.05
21		7.90	2.36	2.10	1.41
20		0	0	0	1.27
19		4.58	4.37	4.61	4.15
18	1	2.19	3.57	4.15	0
17		0	0.60	1.78	3.09
16		2.50	2.60	3.17	2.97
15		0	1.97	2.61	1.96
14		1.62	0	0	0
Sum		26	32	35	30

Note: 1: present band, 0: absent band.

Table 3b. SDS-PAGE electrophoretic profile of Pseudomonas aeruginosa with intensities after treating with silver nanoparticles.

Molecular weight (kDa)	Marker	Control	1%	3%	5%
90		0	3.05	0.11	0
88		4.21	0	0	0
84		4.33	0	0	0
78		2.13	0	0	0
76		0.72	0	0	0
75		0.78	0	0	0
74		0.55	0	0	0
73		0.79	0	0	0
71		1.55	0	0	0
69		0	0	2.35	0
66	1	0	0	0	0
65		4.58	0	0	0
64		1.36	0	0	0
61		3.76	0	0	0
58		1.63	0	15.33	0
56		1.25	0	3.62	0
55		1.03	0	0	0
53		1.26	0	0	0
51		4.49	0	0	0
50		0	25.3	0	0
48		0	0	15.68	0
47		3.97	0	0	0
45	1	0	9.63	0	0
42		5.27	2.90	7.61	0
41		0	0	5.31	0
40		0	4.34	0	0
39		11.33	0	0	0
38		0	3.47	0	0
37		0	0	5.84	0
36		7.56	3.52	0	0
35		0	0	2.43	0
34		3.23	2.20	0	0
33		0	2.51	9.31	0
31		0.92	4.63	0	0
28		6.22	5.95	0	0
27		0	0	7.75	0
25	1	1.29	2.03	0	0
24		3.82	3.66	5.54	0
22		6.13	1.69	4.38	0
21		1.29	1.84	0	0
20		1.40	2.67	4.42	0
19		4.31	1.58	1.94	0
18	1	0	2.65	2.53	0
17		3.83	1.43	1.48	49.0
16		0	2.23	0	0
15		2.39	4.11	1.85	50.99
14		1.57	2.97	1.75	0
sum		32	22	19	2

Note: 1: present band, 0: absent band.

Table 4. RAPD-PCR and ISSR-PCR fragments, distribution of unique and polymorphic bands, monomorphic bands and percentage of polymorphism generated in Staphylococcus aureus treated with silver nanoparticles.

Molecular type	Primer code	Nucleotide sequence 5′ to 3′ of the used primer	Total number of bands	Monomorphic bands	Unique bands	Polymorphic bands	Polymorphism (%)
RAPD	OP-A19	CCTTGACGCA	6	6	0	0	0
	OP-B04	GATGACCGCC	5	1	1	3	80
	OP-C04	CCGCATCTAC	5	4	1	0	20
	OP-C12	CAGCACCCAC	3	3	0	0	0
	OP-Q18	GGGAGCGAGT	6	6	0	0	0
	OP-Z03	TCGGATCCG T	8	1	2	5	87.5
Total			33	21	4	8
Mean			5.5	3.5	0.7	1.3	31.3
ISSR	14A	CTCCTCCTCGC	4	3	0	1	25
	44B	CTCTCTCTCTCTCTCTTG	5	2	2	1	60
	HB-08	GAGAGAGAGAGAGG	6	3	1	2	50
	HB-11	GTGTGTGTGTGTCC	6	5	0	1	16.7
	HB-12	CACCACCACGC	6	2	2	2	66.7
Total			27	15	5	7
Mean			5.4	3	1	1.4	43.7

Table 5. RAPD-PCR and ISSR-PCR fragments, distribution of unique and polymorphic bands, monomorphic bands and percentage of polymorphism generated in Pseudomonas aeruginosa treated with silver nanoparticles.

Molecular type	Primer code	Nucleotide sequence 5′ to 3′ of the used primer	Total number of bands	Monomorphic bands	Unique bands	Polymorphic bands	Polymorphism (%)
RAPD	OP-A19	CCTTGACGCA	7	6	1	0	14.28
	OP-B04	GATGACCGCC	7	0	3	4	100
	OP-C04	CCGCATCTAC	6	4	0	2	33.33
	OP-C12	CAGCACCCAC	4	2	0	2	50
	OP-Q18	GGGAGCGAGT	9	5	2	2	44.44
	OP-Z03	TCGGATCCGT	6	0	2	4	100
Total			39	17	8	14
Mean			6.5	2.8	1.3	2.3	57
ISSR	14A	CTCCTCCTCGC	5	2	1	2	60
	44B	CTCTCTCTCTCTCTCTTG	7	2	4	1	71.42
	HB-08	GAGAGAGAGAGAGG	4	1	0	3	75
	HB-11	GTGTGTGTGTGTCC	9	4	4	1	55.55
	HB-12	CACCACCACGC	4	4	0	0	0
Total			29	13	9	7
Mean			5.8	2.6	1.8	1.4	52.4

Figure 9. UPGMA dendogram based on (a) RAPD-PCR and (b) ISSR-PCR analysis, clarifying genetic differences between the studied treatments: (1) Staphylococcus aureus; (2) Pseudomonas aeruginosa.