Cytogenotoxicity potentials of cocoa pod and bean-mediated green synthesized silver nanoparticles on Allium cepa cells

Figures & data

Figure 1. Synthesis of biogenic AgNPs using cocoa pod and seed extracts.

Figure 2. UV-vis absorption spectra of biogenic synthesized AgNPs using extracts of cocoa pod husk and seed.

Figure 3. (a) TEM micrographs; (b) SAED patterns; and (c) EDX spectra of the biosynthesized AgNPs.

Table 1. Cytogenotoxic effects of cocoa extract mediated silver nanoparticles on Allium cepa cells at 24 h exposure.

Conc. (μg ml^–1)	No. of dividing cells	Mitotic index (%)	Mitotic inhibition (%)	Prophase	Metaphase	Anaphase	Telophase	CB	CM	VC	SC	FG	No. of aberration	% aberrant per cell scored
Control	512	10.24^a	–	33.20	25.78	20.51	20.51	–	–	–	–	–	–	–
CPHE-AgNPs
0.01	226	4.52^b#	27.33	43.36	26.55	16.37	13.72	–	3	–	3	–	6	0.12
0.1	203	4.06^c#	34.72	42.86	27.59	14.78	14.78	–	3	1	4	–	8	0.16
1	153	3.06^d#	50.80	40.52	25.49	25.49	15.03	–	4	1	3	–	8	0.16
10	117	2.34^e#	62.37	41.03	26.50	19.66	12.82	–	4	–	3	–	7	0.14
100	48	0.96^f#	84.56	43.75	25.00	14.58	16.67	–	4	–	5	–	9	0.18
CBE-AgNPs
0.01	286	5.72^b	7.74	37.72	27.27	18.88	15.73	–	3	1	4	–	8	0.16
0.1	210	4.20^c#	32.25	38.10	25.71	19.52	16.67	3	3	–	8	–	14	0.28
1	192	3.84^d#	38.06	40.63	26.56	16.67	16.15	2	3	2	10	–	17	0.34
10	147	2.94^e#	52.58	41.50	25.85	18.37	14.29	–	2	–	1	–	3	0.06
100	48	0.96^f#	84.51	43.75	27.08	12.50	16.67	–	–	–	9	–	9	0.18
Ags
0.01	398	7.96^b	17.94	26.63	24.37	25.38	23.62	–	2	–	2	–	4	0.08
0.1	376	7.52^b	22.47	26.60	24.47	24.74	24.20	1	1	–	1	–	3	0.06
1	289	5.78^c	40.41	29.07	24.22	24.91	21.80	2	1	–	1	–	4	0.08
10	254	5.08^d#	47.62	29.92	24.80	22.83	22.44	1	1	–	2	2	6	0.12
100	232	4.64^e#	52.16	28.88	24.57	22.84	23.71	–	1	–	2	2	5	0.10

Notes:

5000 cells were scored per concentration; CB: chromosomal bridge; CM: c-mitosis; VC: vagrant chromosome; SC: sticky chromosome; FG: fragmentation.

Variation in alphabets along column indicate significant difference (p < 0.05) using ANOVA and Duncan test.

# Mitotic index of the test concentration less than one half of the value obtained for the control.

Figure 4. Mitotic stages and chromosomal aberrations observed for A. cepa root cells treated with biogenic synthesized AgNPs and Ags.

Table 2. Cytogenotoxic effects of cocoa extract mediated silver nanoparticles on Allium cepa cells at 48 h exposure.

Conc. (μg ml^–1)	No. of dividing cells	Mitotic index (%)	Mitotic inhibition (%)	Prophase	Metaphase	Anaphase	Telophase	CB	CM	VC	SC	FG	No. of aberration	% aberrant per cell scored
Control	515	10.30 ^a	–	32.43	22.33	21.75	23.50	–	–	–	–	–	–	–
												–
CPHE-AgNPs
0.01	273	5.46^b	9.30	38.10	27.47	19.05	15.38	–	1	–	5	–	6	0.12
0.1	243	4.86^c#	19.26	37.45	27.57	19.75	15.23	–	6	2	9	–	17	0.34
1.0	190	3.80^d#	36.87	36.84	28.42	19.47	15.26	–	6		14	–	20	0.40
10.0	137	2.74^e#	54.48	37.23	27.01	18.25	17.52	1	8	2	13	–	24	0.48
100	–	–	–	–	–	–	–	–	–	–	–	–	–	–
CBE-AgNPs
0.01	267	5.34^b	11.29	41.20	27.72	17.60	13.48	–	–	–	4	–	4	0.08
0.1	224	4.48^c#	25.58	35.27	26.34	20.54	17.86	–	4	2	4	–	10	0.20
1	188	3.76^d#	37.54	39.36	26.06	19.15	15.43	–	4	3	9	–	16	0.32
10	112	2.44^e#	59.46	39.29	25.89	20.54	14.29	–	3	1	14	–	19	0.38
100	–	–	–	–	–	–	–	–	–	–	–	–	–	–
Ags
0.01	474	9.48^a	6.68	31.22	23.21	23.00	22.57	1	–	–	1	–	2	0.04
0.1	449	8.98^ab	11.78	30.73	24.05	23.39	21.83	–	1	–	3	–	5	0.10
1	363	7.26 ^b	28.68	26.72	24.52	25.07	23.69	1	2	–	1	–	4	0.08
10	276	5.52 ^c	45.77	28.26	24.64	23.91	23.19	2	2	–	2	–	7	0.14
100	229	4.58 ^d#	55.01	27.51	23.58	23.58	25.33	3	1	–	4	–	8	0.16

Notes:

5000 cells were scored per concentration; SC: sticky chromosome; CB: chromosomal bridge; CM: c-mitosis; VC: vagrant chromosome; FG: fragmentation.

Variation in alphabets along column indicate significant difference (p < 0.05) using ANOVA and Duncan test.

# Mitotic index of the test concentration less than half of the value obtained for the control.

Table 3. Cytogenotoxic effects of cocoa extracts mediated silver nanoparticles on Allium cepa cells at 72 h exposure.

Conc. (μg ml^–1)	No. of dividing cells	Mitotic index (%)	Mitotic inhibition (%)	Prophase	Metaphase	Anaphase	Telophase	CB	CM	VC	SC	FG	No. of aberration	% aberrant per cell scored
Control	494	9.88 ^a	–	31.58	24.49	20.65	23.28	–	–	–	–	–	–	–
CPHE-AgNPs
0.01	226	4.52^b#	17.81	36.28	29.20	19.47	15.04	–	–	–	3	–	3	0.06
0.1	175	3.50^c#	36.36	37.71	30.29	18.29	13.71	–	3	1	10	–	14	0.28
1	142	2.84^d#	48.36	36.62	26.06	21.13	16.20	1	4	1	9	–	15	0.30
10	96	1.92^e#	65.09	39.58	25.00	24.00	11.46	3	9	2	10	–	24	0.48
100	–	–	–	–	–	–	–	–	–	–	–	–	–	–
CBE-AgNPs
0.01	248	4.96^b	9.81	35.89	27.82	21.37	14.92	–	3	–	5	–	8	0.16
0.1	186	3.72^c#	32.36	37.10	26.88	20.43	15.59	–	3	6	8	–	17	0.34
1	150	3.00^d#	45.45	36.00	25.33	22.67	16.00	–	6	4	8	–	18	0.36
10	97	1.94^e#	64.72	40.21	28.87	18.56	13.40	3	6	4	9	–	22	0.44
100	–	–	–	–	–	–	–	–	–	–	–	–	–	–
Ags
0.01	392	7.84^b	20.0	27.04	25.26	23.72	23.98	3	1	–	2	–	6	0.12
0.1	350	7.00 ^b	28.57	28.29	25.71	23.71	22.29	4	2	–	2	1	9	0.18
1	239	4.78 ^c#	51.22	28.03	23.43	22.59	25.94	–	2	–	5	–	7	0.14
10	221	4.42 ^cd#	54.89	28.96	23.98	22.62	24.43	1	3	–	4	–	8	0.16
100	198	3.96 ^d#	59.59	30.81	24.75	23.23	21.21	4	3	–	2	–	9	0.18

Notes:

5000 cells were scored per concentration; SC: sticky chromosome; CB: chromosomal bridge; CM: c-mitosis; VC: vagrant chromosome; FG: fragmentation.

Variation in alphabets along column indicate significant difference (p < 0.05) using ANOVA and Duncan test.

# Mitotic index of the test concentration less one than half of the value obtained for the control.

Figure 5. Variations in root numbers of A. cepa treated with cocoa pod and seed extract mediated AgNPs.

Note: Different letters and numbers indicate significant difference between control and group of treatment concentrations of AgNPs (p < 0.05). Each Bar is separated into upper and lower quartile by the mean at the centre. Whiskers represent maximum and minimum root numbers.

Table 4. Toxic effects of cocoa extract mediated green synthesized AgNPs on A. cepa root growth.

Concentration (μg ml^–1)	Root length CPHE-AgNPs	PRI (%)	PRLRC (%)	Root length CBE-AgNPs	PRI (%)	PRLRC (%)	Root length Ags	PRI (%)	PRLRC (%)
0	4.07^a±0.07	–	100	4.07^a±0.07	–	100	4.07^a±0.07	–	100
0.01	4.08^a±0.07	0.72	99.27	3.59^b ±0.10	13.07	86.92	3.03^b±0.91	25.56	74.44
0.10	3.28^b±0.06	20.19	79.80	3.47^b ±0.11	15.98	84.01	2.76^c ±0.61	32.19	67.81
1.0	2.50^c±0.05	39.17	60.82	3.36^b ±0.06	18.64	81.35	2. 32^c ±0.63	43.00	57.00
10.0	0.53^d±0.01	87.10	12.89	0.86^c ±0.03	79.17	20.82	1.58^d±0.43	61.18	38.82
100.0	0.13^e±0.00	96.83	3.16	0.19^d ±0.01	95.39	4.60	0.40^e±0.02	90.17	9.83

Note:

PRI: percentage root inhibition; PRLRC: percentage root length relative to control.

Different letters in a column indicate significant difference (p < 0.05) using ANOVA and Duncan test.

Figure 6. Relationship of root growth response to different biogenic AgNPs.