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Biochemical‑, Biophysical‑, and Microarray-Based Antifungal Evaluation of the Buffer-Mediated Synthesized Nano Zinc Oxide: An in Vivo and in Vitro Toxicity Study
journal contribution
posted on 2016-02-20, 04:13 authored by Prasun Patra, Shouvik Mitra, Nitai Debnath, Arunava GoswamiHere we describe a simple, novel method of zinc oxide
nanoparticle
(ZNP) synthesis and physicochemical characterization. The dose-dependent
antifungal effect of ZNPs, compared to that of micronized zinc oxide
(MZnO), was studied on two pathogenic fungi: Aspergillus niger and Fusarium oxysporum. Superoxide dismutase (SOD)
activity, ascorbate peroxidase activity, catalase activity, glutathione
reductase (GR) activity, thiol content, lipid peroxidation, and proline
content in ZNP-treated fungal samples were found to be elevated in
comparison to the control, which strongly suggested that the antifungal
effect of ZNPs was due to the generation of reactive oxygen species
(ROS). Protein carbonylation, another marker of oxidative stress,
was also evaluated by the dinitrophenyl hydrazine (DNPH) binding assay
and Fourier transform infrared (FTIR) spectral analysis followed by
Western blot and microarray analysis of fungal samples to confirm
ROS generation by ZNPs. Micrographic studies for the morphological
analysis of fungal samples (ZNP-treated and a control) exhibited an
alteration in fungal morphology. The bioavailability of ZNPs on fungal
cell was confirmed by energy-dispersive X-ray (EDX) analysis followed
by high-resolution transmission electron microscopy (HR-TEM) and confocal
microscopic analysis of the fungal samples. In vivo acute oral toxicity,
acetylcholine esterase activity, and a fertility study using a mice
model were also investigated for ZNPs. The long-term toxicity of ZNPs
through intravenous injection was evaluated and compared to that of
MZnO. The in vitro comparative toxicity of ZNPs and MZnO was evaluated
on MRC-5 cells with the help of water-soluble tetrazolium (WST-1)
and lactate dehydrogenase (LDH) assays. These results suggested that
ZNPs could be used as an effective fungicide in modern medical and
agricultural sciences.
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Keywords
ZNPbinding assayGRMRCMicrographic studiesSuperoxide dismutaseFTIRmice modelmicroarray analysismicronized zinc oxidelipid peroxidationproline contentLDHzinc oxide nanoparticleWestern blotantifungal effectVitro Toxicity StudyHerereactive oxygen speciesdinitrophenyl hydrazinelactate dehydrogenasethiol contentphysicochemical characterizationFusarium oxysporumascorbate peroxidase activityProtein carbonylationacetylcholine esterase activitytransmission electron microscopyAspergillus nigercatalase activityoxidative stressEDXWSTglutathione reductasesampleSODDNPHROS generationnovel methodfertility study
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