Abstract
In this work, we present an ecofriendly, non-hazardous, green synthesis of zinc oxide nanoparticles (ZnO NPs) by leaf extract of Crotalaria verrucosa (C. verrucosa). Total phenolic content, total flavonoid and total protein contents of C. verrucosa were determined. Further, synthesized ZnO NPs was characterized by UV–visible spectroscopy (UV-vis), X-ray diffractometer (XRD), Fourier transform infra-red (FTIR) Spectra, transmission electron microscope (TEM), and Dynamic light scattering (DLS) analysis. UV-vis shows peak at 375 nm which is unique to ZnO NPs. XRD analysis demonstrates the hexagonal phase structures of ZnO NPs. FTIR spectra demonstrates the molecules and bondings associated with the synthesized ZnO NPs and assures the role of phytochemical compounds of C. verrucosa in reduction and capping of ZnO NPs. TEM image exhibits that the prepared ZnO NPs is hexagonal shaped and in size ranged between 16 to 38 nm which is confirmed by DLS. Thermo-gravimetric analysis (TGA) was performed to determine the thermal stability of biosynthesized nanoparticles during calcination. The prepared ZnO NPs showed significant antibacterial potentiality against Gram-positive (S. aureus) and Gram-negative (Proteus vulgaris, Klebsiella pneumoniae, and Escherichia coli) pathogenic bacteria and SEM image shows the generalized mechanism of action in bacterial cell after NPs internalization. In addition, NPs are also found to be effective against the studied cancer cell lines for which cytotoxicity was assessed using MTT assay and results demonstrate highest growth of inhibition at the concentration of 100 µg/mL with IC50 value at 7.07 µg/mL for HeLa and 6.30 µg/mL for DU145 cell lines, in contrast to positive control (C. verrucosa leaf extract) with IC50 of 22.30 µg/mL on HeLa cells and 15.72 µg/mL on DU145 cells. Also, DAPI staining was performed in order to determine the effect on nuclear material due to ZnO NPs treatment in the studied cell lines taking leaf extract as positive control and untreated negative control for comparison. Cell migration assay was evaluated to determine the direct influence of NPs on metastasis that is potential suppression capacity of NPs to tumor cell migration. Outcome of the synthesized ZnO NPs using C. verrucosa shows antimicrobial activity against studied microbes, also cytotoxicity, apoptotic mediated DNA damage and antiproliferative potentiality in the studied carcinoma cells and hence, can be further used in biomedical, pharmaceutical and food processing industries as an effective antimicrobial and anti-cancerous agent.
Highlights
Nanoparticles (NPs) are the engineered assemblage of materials having nanoscale dimensions with a size distribution range of 1–100 nm diameters as defined by the National Nanotechnology Initiative (NNI), with unique physicochemical properties due to their nanosize, hydrophilic and hydrophobic nature, functional moieties on surface, high surface to volume ratio and aggregation properties [1].Nanoparticles permit precision engineering to track and regulate their interaction with biological systems and nanotechnology integrated with biology is very much involved in the creation of modern and more effective antimicrobial and anticancerous agents
Ethanolic extract of C. verrucosa has been previously examined for its phytochemical tests and confirmed the presence of flavonoids and phytosterols in this plant
C. verrucosa was subjected to the determination of phytochemicals such as polyphenols, flavonoids and proteins
Summary
Nanoparticles permit precision engineering to track and regulate their interaction with biological systems and nanotechnology integrated with biology is very much involved in the creation of modern and more effective antimicrobial and anticancerous agents. Metal oxide nanoparticles provide promising and wide perspectives for biomedical fields through the production of nanomaterials, notably antimicrobial, anticancerous drugs, gene delivery, cell imaging, and biosensing [2]. Mutations in bacterial genes and multiple drug resistance (MDR) have raised challenges to treat MDR microorganisms because resistance drastically restricts therapeutic options and there is a need for effective antibacterial agents without adverse effects on treated cells. ZnO NPs are efficient multifunctional inorganic metal oxide NPs that have a wide energy gap of 3.37 eV with short wavelength and large exciton binding energy of 60 meV, is considered to be moderately toxic but is extremely reactive and sensitive with extreme absorbent potentiality [3]. Zinc is considered to be a significant trace element in tissues of the body, such as brain, muscle, bone, and skin, maintaining many physiological functions and without which few enzymes namely carbonic anhydrase, carboxypeptidase, and alcohol dehydrogenase become quiescent [4]
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