Green Synthesized FM-AgNPs Lead to Alterations in Hematology, Oxidative Stress Biomarkers, and Microanatomy of Liver and Spleen in Rats

Rubab, Moazzam; Zain, Ali; Mubeen, Bismillah; Tariq, Huma; Malik, Arif; Arshad, Najma

doi:10.1590/1678-4324-2023220098

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Article - Human and Animal Health • Braz. arch. biol. technol. 66 • 2023 • https://doi.org/10.1590/1678-4324-2023220098 copy

Green Synthesized FM-AgNPs Lead to Alterations in Hematology, Oxidative Stress Biomarkers, and Microanatomy of Liver and Spleen in Rats

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Widespread use of green synthesized silver nanoparticles (AgNPs) in pharmaceutical and cosmetics demand for its toxicological assessment. The current study describes the influence of exposure to kumquat, Fortunella margarita (FM) synthesized AgNPs in albino rats. Animals (n=60) were randomly distributed into four groups. Group, I served as control while groups II-IV were exposed to FM-AgNPs by the single intraperitoneal dose of 1, 5, and 10 mg/kg body weight, respectively. Animals were sacrificed at two-week intervals up to 5 weeks. Hematology, tissue (liver and spleen) oxidative stress biomarkers (SOD, CAT, and MDA), and histopathological examination were performed. Significant elevation in hemoglobin (Hb), Erythrocyte count (RBC), hematocrits (HCT), and hematological indices and depletion in total leukocytes count (TLC) was recorded at different time points. Similarly, a significant increase in SOD, CAT, and MDA activity was observed in both liver and spleen. But changes (p ≤ 0.005) in the spleen was recorded more consistently. Exposures to the high dose of FM-AgNPs lead to significant alterations in microanatomy of the liver and spleen. In liver vacuolization, enucleation, necrosis, congestion, and reduction in the size of cell, nucleus, portal vein, and elevation in cell density per unit area were recorded. In the spleen, except for enlargement in splenic sinuses, no other alterations could be recorded. It is concluded that FM-AgNPs leaves time and dose-dependent toxicity in rats but variations are more evident at high doses.

Keywords:
Fortunella margarita; silver nanoparticles; hematology; oxidative stress biomarkers; histopathology.

HIGHLIGHTS

• Green synthesized silver nanoparticles are not always safe for use.

• Fortunella margarita (FM-AgNPs) lead to alteration in hematological indicators.

• FM-AgNPs leads to elevation in SOD, CAT and MDA in liver and spleen.

• High doses of FM-AgNPs lead to degenerative changes in microanatomy of liver.

Parameter	Week	Group												P F-test
Parameter	Week	Control 0 mg/kg			Low 1 mg/kg			Medium 5 mg/kg			High 10 mg/kg			P F-test
Hb (g/dl)	1	12.60	±	0.06	13.43	±	0.07^*	12.50	±	0.06	12.47	±	0.20	0.01
	3	12.47	±	0.03	13.30	±	0.21	14.57	±	0.03^*	12.83	±	0.55	0.00
	5	12.60	±	0.06	13.40	±	0.17^*	13.40	±	0.06^*	12.50	±	0.12	0.00
RBC (×10⁶/µL)	1	7.35	±	0.01	9.30	±	0.12^*	7.90	±	0.06^*	7.93	±	0.09^*	0.00
	3	7.39	±	0.01	7.87	±	0.09^*	8.77	±	0.07^*	7.27	±	0.15	0.00
	5	7.33	±	0.01	7.73	±	0.15	8.00	±	0.29	7.57	±	0.15	0.13
HCT (%)	1	41.00	±	0.58	48.27	±	0.15^*	45.33	±	0.12^*	41.67	±	0.33	0.00
	3	39.33	±	1.45	46.00	±	1.15^*	47.50	±	0.44^*	41.33	±	0.33	0.00
	5	37.00	±	3.51	40.30	±	0.17	44.47	±	0.87^*	43.53	±	0.29	0.07
TLC (×10³/µL)	1	14.00	±	0.58	8.60	±	0.17^*	11.10	±	0.06^*	9.27	±	0.15^*	0.00
	3	12.00	±	0.58	9.93	±	0.03^*	12.50	±	0.12	10.53	±	0.29^*	0.00
	5	14.00	±	1.15	6.27	±	0.15^*	8.33	±	0.15^*	8.47	±	0.29^*	0.00
NP (%)	1	14.00	±	0.58	22.00	±	0.58^*	14.00	±	1.15	20.13	±	0.88^*	0.00
	3	12.33	±	0.88	17.33	±	0.88^*	20.00	±	0.58^*	25.00	±	1.15^*	0.00
	5	11.00	±	1.15	15.33	±	0.33^*	18.00	±	0.00^*	23.00	±	1.15	0.00
LC (%)	1	86.33	±	0.33	77.33	±	0.33^*	86.00	±	2.31	90.33	±	0.88	0.00
	3	84.00	±	0.58	83.00	±	2.31	80.33	±	0.33	75.00	±	2.89^*	0.03
	5	84.33	±	0.33	85.00	±	2.89	70.00	±	2.89^*	67.33	±	0.33	0.00
MCV (fL)	1	56.00	±	0.58	50.33	±	2.91	55.67	±	0.88	54.00	±	0.58	0.11
	3	53.00	±	1.15	59.00	±	2.31^*	54.47	±	0.29	66.00	±	0.58^*	0.00
	5	55.00	±	0.58	52.00	±	0.00	55.47	±	0.29	57.33	±	1.45	0.01
MCH (pg)	1	17.00	±	0.58	14.00	±	0.58	16.00	±	1.73	16.00	±	0.58	0.26
	3	17.00	±	0.58	17.00	±	1.15	16.57	±	0.30	17.00	±	0.00	0.95
	5	18.67	±	0.88	17.13	±	0.19	16.60	±	0.31	16.00	±	0.58^*	0.04
MCHC (g/dL)	1	31.00	±	1.15	28.10	±	0.49	28.00	±	1.73	27.47	±	0.52	0.17
	3	28.00	±	0.58	28.00	±	0.58	30.67	±	0.49	31.03	±	1.13^*	0.03
	5	28.00	±	0.58	33.27	±	0.32^*	30.10	±	0.62	28.03	±	0.55	0.00

Parameter	Week	Group												P F-test
Parameter	Week	Control 0 mg/kg			Low 1 mg/kg			Medium 5 mg/kg			High 10 mg/kg			P F-test
CAT (Umin^-1g^-1)	1	6.12	±	0.27	7.14	±	0.15	7.12	±	0.09	7.59	±	0.49^*	0.04
	3	6.04	±	0.19	7.35	±	0.33	6.29	±	0.60	7.55	±	0.30	0.05
	5	6.30	±	0.43	6.49	±	0.31	7.39	±	1.16	6.92	±	0.70	0.72
SOD (Ug^-1)	1	27.97	±	0.87	45.86	±	3.00^*	50.29	±	0.85^*	51.86	±	6.23^*	0.00
	3	26.33	±	0.84	47.65	±	3.67^*	40.38	±	5.17^*	55.46	±	2.02^*	0.00
	5	31.52	±	0.63	36.89	±	1.04	36.27	±	2.47	45.85	±	2.31^*	0.00
MDA (µMg^-1)	1	8.44	±	0.36	9.42	±	0.27	8.42	±	0.27	8.67	±	0.23	0.12
	3	7.96	±	0.08	8.16	±	0.18	7.06	±	0.52	9.42	±	0.36^*	0.00
	5	9.24	±	0.34	7.63	±	0.14	9.42	±	0.40	12.29	±	2.69	0.19

Parameter	Week	Group												P F-test
Parameter	Week	Control 0 mg/kg			Low 1 mg/kg			Medium 5 mg/kg			High 10 mg/kg			P F-test
CAT (Umin^-1g^-1)	1	42.32	±	0.78	90.76	±	1.07^*	78.07	±	1.78^*	70.13	±	0.88^*	0.00
	3	56.62	±	1.13	58.82	±	1.70	59.75	±	2.59	65.35	±	0.99^*	0.03
	5	35.64	±	0.45	66.71	±	1.02^*	42.32	±	1.48^*	70.95	±	2.67^*	0.00
SOD (Ug^-1)	1	212.64	±	1.18	362.25	±	5.46^*	566.31	±	0.87^*	467.33	±	5.27^*	0.00
	3	268.36	±	4.62	368.79	±	0.51^*	238.12	±	2.67^*	343.47	±	4.05^*	0.00
	5	190.93	±	2.63	417.22	±	2.53^*	375.94	±	2.88^*	354.27	±	2.57^*	0.00
MDA (µMg^-1)	1	69.22	±	1.35	245.34	±	31.04^*	152.86	±	23.06^*	217.46	±	11.85^*	0.00
	3	79.34	±	3.03	275.66	±	10.04^*	161.13	±	7.02	258.13	±	21.88^*	0.00
	5	80.90	±	5.51	308.53	±	15.61^*	191.08	±	7.94^*	275.80	±	14.11^*	0.00

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