A pilot toxicology study of biogenic silver nanoparticles: in vivo by intraperitoneal and intravenous infusion routes in rats

Figures & data

Table 1. Intravenous route of administration of three different concentrations for Bio-AgNPs: Experiment (A).

Group: Bio-AgNPs	Dose concentration in (µg/kg)
Group B: (NOAEC)	124
Group C: (IC50)	128
Group D: (TLC)	132

Bio-AgNPs: biogenic silver nanoparticles; NOAEC: no observable adverse effect concentration; IC50: inhibitory concentration; TLC: total lethal concentration.

Table 2. Intraperitoneal route of administration of three different concentration for Bio-AgNPs: Experiment (B).

Group : Bio-AgNPs	Dose concentration in (µg/kg)
Group B: (NOAEC)	97.64
Group C: (IC50)	102.8
Group D: (TLC)	105.47

Bio-AgNPs: biogenic silver nanoparticles; NOAEC: no observable adverse effect concentration; IC₅₀: inhibitory concentration; TLC: total lethal concentration.

Table 3. Haematology study for rat treated with biogenic nanoparticles through intravenous and intraperitoneal route of administration after 14th and 28th day of observation.

Parameters	Group: Bio-AgNPs 14th day			Group: Bio-AgNPs 28th day			Control	Ref. range
	NOAEC	IC50	TLC	NOAEC	IC50	TLC
WBC								4–10
i/v	7.8 ± 0.4	6.2 ± 0.5	5.8 ± 0.56	7.2 ± 0.089	6.4 ± 0.8	6.52 ± 0.71	9.71 ± 0.98
i/p	9.43 ± 0.63	8.86 ± 0.836	2.36 ± 0.053	8.3 ± 0.072	5.7 ± 0.038	7.4 ± 0.096	9.82 ± 0.094
RBC								3.50–5.50
i/v	4.64 ± 0.07	5.02 ± 0.04	5.61 ± 0.064	5.16 ± 0.45	5 ± 0.071	5.17 ± 0.38	4.98 ± 0.072
i/p	5.24 ± 0.054	4.30 ± 0.35	3.11 ± 0.84	5.24 ± 0.031	5.78 ± 0.023	4.56 ± 0.092	4.67 ± 0.053
Hb								110–160
i/v	136 ± 3.2	141 ± 5.9	140 ± 6.5	133 ± 7.4	140.9 ± 5.75	142.6 ± 4.9	127 ± 5.32
i/p	128 ± 4.94	140 ± 6.63	158 ± 4.66	134.8 ± 5.4	145.6 ± 4.83	147.2 ± 5.22	132 ± 4.054
Platelet count								100–300
i/v	230 ± 4.7	248.8 ± 6.46	235 ± 6.3	243 ± 5.86	248.8 ± 5.46	235 ± 6.3	232 ± 5.3
i/p	302 ± 5.39	402.8 ± 6.65	587 ± 7.38	280.2 ± 5.41	295 ± 4.72	312.6 ± 5.7	245 ± 4.5
Clotting time								2–5
i/v	4.5 ± 0.085	4.7 ± 0.0015	4.7 ± 0.0065	4.8 ± 0.04	4.7 ± 0.082	4.5 ± 0.9	6.6 ± 0.012
i/p	4.9 ± 0.054	1.8 ± 0.003	0.5 ± 0.094	5 ± 0.08	5 ± 0.05	5.03 ± 0.76	5.9 ± 0.005

i/v: intravenous route; i/p: intraperitoneal route; Bio-AgNPs: biogenic silver nanoparticles; NOAEC: no observable adverse effect concentration; IC₅₀: inhibitory concentration; TLC: total lethal concentration; WBC: white blood cells (SI unit: 10⁹/L); RBC: red blood cells (SI unit: 10⁹/L); Hb: haemoglobin (SI unit: g/L); Platelet count (SI unit: 10⁹/L); clotting time, in min.

All data are expressed in mean ± SD of the mean of the three independent experiments; each was carried out in triplicate, N = 6 rats per group.

Figure 1. Change in body weight for untreated (control) and treated rats with Bio-AgNPs through (A) intraperitoneal (i/p), (B) intravenous (i/v) route of administration up to 28th day of observation. *All treatments of Bio-AgNPs (Ag) at their total lethal concentration were administered through i/v and i/p and kept in observation for 28 days; N = 6 rats per group.

Table 4. Body weight changes for rat treated with biogenic nanoparticles through intravenous and intraperitoneal route of administration.

Days after injection	Group: Bio-AgNPs Body weight in (g) (i/v)			Group: Bio-AgNPs Body weight in (g) (i/p)			Control
	NOAEC	IC50	TLC	NOAEC	IC50	TLC	Body weight in (g)
1	100 ± 7.3	100 ± 5.3	100 ± 4.3	100 ± 3.53	100 ± 4.89	100 ± 6.3	100 ± 5.56
2	100 ± 6.4	100 ± 7.05	100 ± 6.05	100 ± 4.94	100.5 ± 5.042	100 ± 8.5	100 ± 6.45
4	108 ± 7.43	106 ± 6.63	101 ± 4.7	102.3 ± 5.93	106.6 ± 7.88	100.5 ± 7.2	106 ± 5.51
6	110 ± 5.5	105 ± 5.78	101 ± 4.5	104.2 ± 4.96	108.2 ± 6.65	98.8 ± 5.27	105 ± 5.37
8	110 ± 6.3	107 ± 6.93	102.6 ± 6.93	104 ± 3.73	108 ± 5.32	96.3 ± 7.63	108 ± 6.48
10	112 ± 5.4	108 ± 6.02	101 ± 3.5	106.5 ± 4.895	110 ± 643	96.4 ± 8.32	112 ± 4.76
12	114 ± 6.5	108 ± 5.89	103 ± 4.46	106.8 ± 4.78	110.5 ± 6.85	94.6 ± 7.93	116 ± 5.6
14	114 ± 5.08	106 ± 6.74	101 ± 5.58	107 ± 5.3	108.4 ± 7.54	92.5 ± 6.43	116 ± 6.71
16	114 ± 7.03	107.5 ± 7.085	102.3 ± .5.6	109 ± 6.9	104.2 ± 6.45	96.9 ± 7.64	121 ± 7.56
18	117 ± 6.7	100.5 ± 7.4	103.6 ± 6.5	110 ± 5.74	104 ± 7.32	98.4 ± 6.73	121 ± 8.53
20	117 ± 8.93	101 ± 5.7	105 ± 4.8	110 ± 4.82	103.2 ± 6.75	100.2 ± 8.83	121 ± 5.42
22	118 ± 6.05	102.5 ± 4.5	106.4 ± 7.54	109 ± 7.93	102.3 ± 8.73	102.5 ± 5.54	121 ± 6.34
24	120 ± 6.07	106 ± 06.5	106 ± 6.4	108.6 ± 5.85	102.5 ± 6.94	102 ± 8.63	124 ± 4.3
26	120 ± 7.23	106.9 ± 5.6	106 ± 7.91	112.5 ± 7.09	100.8 ± 7.5	101.8 ± 6.5	124 ± 5.82
28	122 ± 5.78	108 ± 05.56	106 ± 6.85	116 ± 3.32	101.8 ± 8.64	100.2 ± 8.63	124 ± 6.568

i/v: intravenous route; i/p: intraperitoneal route; Bio-AgNPs: biogenic silver nanoparticles; NOAEC: no observable adverse effect concentration; IC₅₀: inhibitory concentration; TLC: total lethal concentration.

All data are expressed in mean ± SD of the mean of the three independent experiments; each was carried out in triplicate, N = 6 rats per group.

Figure 2. Biochemical results for rats treated with Bio-NPs through intraperitoneal and intravenous route of administration at [A] 14th day and [B] 28th day. Note: Ag: Bio-AgNPs, i/p: intraperitoneal, i/v: intravenous. *All data are expressed in mean ± SD of the mean of the three independent experiments; each was carried out in triplicate, N = 6 rats per group.

Table 5. Biochemical assays for rat treated with biogenic nanoparticles through intraperitoneal and intravenous route of administration for 14th and 28th day of observation.

Tests	Group: Bio-AgNPs 14th day			Group: Bio-AgNPs 28th day			Ref. range
	NOAEC	IC50	TLC	NOAEC	IC50	TLC
S. creatinine							0.7–1.4
i/v	0.92 ± 0.021	0.68 ± 0.078	0.62 ± 0.057	0.90 ± 0.063	0.88 ± 0.051	0.85 ± 0.062
i/p	0.85 ± 0.027	0.86 ± 0.053	0.82 ± 0.094	0.91 ± 0.003	0.86 ± 0.078	0.85 ± 0.085
ALT							0.0–40.0
i/v	24.2 ± 2.63	39 ± 4.28	42.7 ± 2.84	28.6 ± 1.92	35.6 ± 2.61	44.2 ± 1.35
i/p	29.3 ± 3.67	40.6 ± 3.97	50 ± 4.45	30.5 ± 1.83	41.4 ± 02.76	48.5 ± 2.64
AST							5.0–34.0
i/v	26.7 ± 3.36	32.8 ± 5.23	34.13 ± 4.29	28.2 ± 3.7	32.3 ± 3.83	34.8 ± 4.82
i/p	28.3 ± 3.95	32.7 ± 3.39	41 ± 2.85	30.5 ± 5.1	34.5 ± 5.2	40.5 ± 6.6
ALP							37–103
i/v	72.6 ± 4.9	80.6 ± 3.92	82.7 ± 2.83	74.8 ± 2.15	82.7 ± 5.86	89.6 ± 4.92
i/p	84.1 ± 2.65	90.7 ± 3.5	105 ± 4.56	86.7 ± 2.73	94.7 ± 3.75	100 ± 3.83

i/v: intravenous route; i/p: intraperitoneal route; NOAEC: no observable adverse effect concentration; IC₅₀: inhibitory concentration; TLC: total lethal concentration; S. creatinine (mg/dl); ALT: alanine aminotransferase (IU/L); AST, aspartate aminotransferase (IU/L); ALP, alkaline phosphatase (IU/L).

All data are expressed in mean ± SD of the mean of the three independent experiments; each was carried out in triplicate, N = 6 rats per group.

Table 6. Weight of organs or organ index (Organ_X) for rat treated with biogenic nanoparticles through intraperitoneal and intravenous route of administration for 14th and 28th day of observation.

Tests	Group : Bio-AgNPs 14th day			Group : Bio-AgNPs 28th day			Control
	NOAEC	IC50	TLC	NOAEC	IC50	TLC
Heart (HX)
i/v	1 ± 0.01	0.97 ± 0.033	1.4 ± 0.054	1.2 ± 0.004	1.5 ± 0.0092	1.4 ± 0.0037	1 ± 0.02
i/p	1 ± 0.0256	0.9 ± 0.067	1.47 ± 0.03	1.4 ± 0.0032	1.21 ± 0.095	1.38 ± 0.004	1.4 ± 0.05
Liver (LiX)
i/v	1.8 ± 0.065	1.6 ± 0.055	1.35 ± 0.07	1.8 ± 0.084	1.68 ± 0.0087	1.77 ± 0.0053	1.8 ± 0.043
i/p	1.2 ± 0.044	1 ± 0.043	2.07 ± 0.054	1.75 ± 0.005	1.2 ± 0.094	2.12 ± 0.007	1.67 ± 0.032
Spleen (SX)
i/v	1.11 ± 0.045	1.2 ± 0.022	0.97 ± 0.06	1.12 ± 0.002	1.2 ± 0.063	1.34 ± 0.004	1.16 ± 0.028
i/p	1.1 ± 0.056	0.8 ± 0.087	0.6 ± 0.09	1.18 ± 0.006	1.05 ± 0.005	0.96 ± 0.05	1.18 ± 0.067
Lungs (LuX)
i/v	2 ± 0.012	1.14 ± 0.023	1.19 ± 0.03	2.2 ± 0.004	1.76 ± 0.034	1.82. ± 0.0063	1.56 ± 0.082
i/p	1 ± 0.043	1.6 ± 0.054	1.56 ± 0.07	1.7 ± 0.0056	1.65 ± 0.006	1.59 ± 0.0049	1.72 ± 0.028
Kidneys (KX)
i/v	2 ± 0.087	1.9 ± 0.077	0.92 ± 0.09	1.96 ± 0.0054	1.9 ± 0.096	1.52 ± 0.006	1.23 ± 0.099
i/p	1 ± 0.017	0.89 ± 0.023	0.84 ± 0.05	1.5 ± 0.0032	1.3 ± 0.006	1.25 ± 0.075	1.52 ± 0.091
Thymus (TX)
i/v	0.171 ± 0.04	0.5 ± 0.067	0.64 ± 0.01	0.173 ± 0.007	0.17 ± 0.094	0.21 ± 0.008	0.18 ± 0.076
i/p	0.182 ± 0.03	0.85 ± 0.021	0.94 ± 0.05	0.179 ± 0.006	0.193 ± 0.003	0.24 ± 0.0098	0.16 ± 0.045
Brain (BX)
i/v	2 ± 0.0076	2.1 ± 0.032	0.98 ± 0.02	2 ± 0.0004	2.4 ± 0.0052	2.2 ± 0.0067	1.89 ± 0.092
i/p	2 ± 0.016	1.3 ± 0.07	1.25 ± 0.066	2.2 ± 0.0056	1.7 ± 0.086	1.2 ± 0.0087	1.72 ± 0.087

All data are expressed in mean ± SD of the mean of the three independent experiments; each was carried out in triplicate, N = 6 rats per group.

Figure 3. H&E staining of rat kidney. (A) Untreated rat for 14th day (−); (B) Untreated rat for 28th day, normal kidney tissues glomerular cells, bowman capsule (−); (C) Rats treated with Bio-AgNPs, i/p route for 14th day (−); (D) Rats treated with Bio-AgNPs, i/p route for 28th day (+); (E) Rats treated with Bio-AgNPs, i/v route for 14th day (+); (F) Rats treated with Bio-AgNPs, i/v route for 28th day (−). *where (−) indicates no changes, (+) indicates mild changes, (++) indicates moderate changes, (+++) indicates severe changes.

Figure 4. H&E staining of rat spleen. (A) Untreated rat for 14th day (−); (B) Untreated rat for 28th day, normal spleen tissues white palp and red palp (−); (C) Rats treated with Bio-AgNPs, i/p route for 14th day (+); (D) Rats treated with Bio-AgNPs, i/p route for 28th day (−); (E) Rats treated with Bio-AgNPs, i/v route for 14th day (−); (F) Rats treated with Bio-AgNPs, i/v route for 28th day (−). *where (−) indicates no changes, (+) indicates mild changes, (++) indicates moderate changes, (+++) indicates severe changes.

Figure 5. H&E staining of rat liver. (A) Untreated rat for 14th day (−); (B) Untreated rat for 28th day, normal liver tissues central vein and sensual (−); (C) Rats treated with Bio-AgNPs, i/p route for 14th day (+); (D) Rats treated with Bio-AgNPs, i/p route for 28th day(−); (E) Rats treated with Bio-AgNPs, i/v route 14th day (+); (F) Rats treated with Bio-AgNPs, i/v route for 28th day (−). *where (−) indicates no changes, (+) indicates mild changes, (++) indicates moderate changes, (+++) indicates severe changes.

Figure 6. H&E staining of rat brain. (A) Untreated rat for 14th day (−); (B) Untreated rat for 28th day (−); (C) Rats treated with Bio-AgNPs, i/p route for 14th day (−); (D) Rats treated with Bio-AgNPs, i/p route for 28th day (−); (E) Rats treated with Bio-AgNPs, i/v route for 14th day (−); (F) Rats treated with Bio-AgNPs, i/v route for 28th day (−). *where (−) indicates no changes, (+) indicates mild changes, (++) indicates moderate changes, (+++) indicates severe changes.