NANOSAFETY OF NANOFERTILIZER AND NANOCARRIER IN RATS MEDIATED THROUGH BREAST MILK
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Date
2019
Authors
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Journal ISSN
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Publisher
ICAR-NDRI, KARNAL
Abstract
Nanotechnology is a promising tool in every aspect of modern life including agriculture. Nanoproducts
like nanofertilizers (nutrients in nanoform) and nanocarriers (nanoporous materials for
controlled release) are becoming centre of attraction due to their role in precision agriculture. There
is dire need of safety assessment before their application due to environmental exposure and
possible health hazard to life forms including humans. Keeping all this in view, the present
investigation was designed to assess the safety of ZnO nanofertilizer (ZNF) and mesoporous silica
nanocarrier (MSN) taking both in vitro and in vivo model into account. Under in vitro study,
cytotoxicity of ZNF and MSN on immune as well as male germ cell was evaluated by MTT, WST8
and neutral red assay. The cell viability in both cases decreased significantly by time (1hr, 4hr) and
concentration (1, 5, 10, 25, 50 and 100 μg/ml) dependent treatment of ZNF compared to control.
The anti-oxidative enzymes (SOD: Superoxide dismutase, GPX: Glutathione peroxidase) increased
significantly (P < 0.01) after 1 hour incubation with 50 μg/ml of ZNF. Lethal Concentration (100
μg/ml) obtained from percentage hemolysis (%hemolysis) assay of RBCs confirms that ZNF is
relative toxic than MSN. In neutrophils and male germ cells treated upto 100 μg/ml MSN no
significant changes was observed in %viability, %hemolysis as well as in the activities of oxidative
stress enzymes as compared to control. Additionally, ZNF at higher concentration leads to
significant loss (p<0.05) in viability and membrane integrity of male germ cell as depicted by
Eosin-Nigrosin assay and hypo-osmotic swelling test, respectively. In case of in vivo study,
lactating nulliparous rats were orally administered 5, 25 and 50mg/kg b.w. ZNF as well as MSN for
consecutively 19 days from the first day of parturition. ZNF was distributed significantly to breast
milk with increasing dose as quantified by inductively coupled plasma mass spectrometry (ICPMS).
Further, ZNF were absorbed by the small intestine and distributed to liver of breast-fed pups.
Histopathology study of intestine and liver of offsprings depicted the gross pathological changes
caused by milk mediated transfer of ZNF. Most of the ZNF and MSN were excreted through feces
of dams. IgA secretion in the intestinal fluid of offsprings increased significantly on increasing dose
of ZNF to dams. Liver injury marker enzymes (AST, ALT) increased significantly in dams and
offspring postnatally. The level of cytokines (TNF-α, IL-1β) in serum increased significantly in the
offsprings of 25mg/kg and 50mg/kg bw group. The total cytochrome P450 reductase and total
antioxidant capacity decreased significantly in the offsprings of 50mg/kg ZNF given dams group.
Further, nitric oxide production and phagocytosis activity of macrophages from offsprings of
50mg/kg ZNF dams group decreased significantly (p<0.05). Real-time polymerase chain reaction
indicated milk mediated expression of anti-oxidative enzymes (SOD, GPX), apoptosis (BAX) and
cytokines (TNFα, IL1β) genes in offspring neutrophils after oral administration of 50/mg kg b.w
ZNF to dams. These results provide information that MSN is comparatively safer than ZNF upto
100 μg/ml concentrations in vitro and 50mg/kg body weight in vivo intubation to dams after
parturition.