Green synthesized zinc oxide nanoparticles induce apoptosis by suppressing PI3K/Akt/mTOR signaling pathway in osteosarcoma MG63 cells

In the recent decade, zinc oxide nanoparticles (ZnO NPs) have been widely explored owing to their versatile properties and prodigious demands in the drug delivery, medical, energy storage, cosmetics, and the healthcare sectors. Therefore, the current work opts for an environmentally benign method to prepare ZnO NPs. The leaf extract of Calendula officinalis L. acts as a reducing agent for the metal ions; therefore, in the current research, ZnO NPs were prepared via green route by using Calendula officinalis leaf extract. Furthermore, the ZnO NPs were analysed with different spectroscopic techniques to confirm the structure and stability of nanomaterials. The prepared ZnO NPs were characterized by XRD, FE-SEM, FT-IR and UV–Vis studies. Also, the antioxidant and antimicrobial properties of the synthesized ZnO NPs were investigated. The XRD result of synthesized ZnO NPs showed the crystalline size 28.23 nm with wurtzite hexagonal structure along with the most intense peak (101). Following preliminary confirmations of the intended ZnO NPs, both big and small agglomerated forms were observed in the FE-SEM, which is often used to determine their exterior assembly. Further, the results of Fourier transform infrared spectroscopy (FT-IR) indicated the formation of pure ZnO NPs with an absorption peak of the Zn–O bond between 4000 cm−1 and 500 cm−1 and no discernible peak in the monitoring range. The UV–Vis spectrum of the green synthesized ZnO NPs were revealed two prominent absorption peaks at 355 nm and 370 nm with energy band gap of 2.986 eV. Using the 1, 1-di phenyl-2-picrylhydrazyl (DPPH) test, the antioxidant activity of the described ZnO NPs was assessed. It demonstrated how, ZnO NPs significantly increased their antioxidant activity by scavenging 1, 1-di phenyl-2-picrylhydrazyl (DPPH) free radicals. It could be seen that synthesis of the naturally occurring plant product ZnO NPs have been acting as an alternate of chemical antioxidant. The antimicrobial analysis was also performed with the help of disk diffusion method where three multi-drug resistant human pathogens namely Staphylococcus aureus, Klebsiella pneumoniae and E.coli were used. The Zone of Inhibition diameter values are 35.2 mm ± 0.9, 23.6 mm ± 0.1 and 13.5 mm ± 0.1, respectively, which showed that the ZnO NPs was highly effective against S. aureus. Thus, the green synthesis method of ZnO NPs using leaf extract of Calendula officinalis is evidence that it is superior and environmentally friendly method for the preparation of ZnO NPs and hence it can be utilized in various nano-medicine approaches.

Green synthesis of zinc oxide nanoparticles using Cinnamomum camphora (L.) Presl leaf extracts and its antifungal activity

Cytotoxicity of green synthesized zinc oxide nanoparticles using Musa acuminata on Vero cells

Potential usage of biosynthesized zinc oxide nanoparticles from mangosteen peel ethanol extract to inhibit Xanthomonas oryzae and promote rice growth

Cancer is a leading cause of death worldwide and sustained focus is on the discovery and development of newer and better tolerated anticancer drugs, especially from plants. In the present study, a simple, eco-friendly, and inexpensive approach was followed for the synthesis of zinc oxide nanoparticles (ZnO NPs) using the aqueous leaf extract of Eclipta prostrata. The synthesized ZnO NPs were characterized by UV-visible absorption spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX), High-resolution transmission electron microscopy (HRTEM), and Selected area (electron) diffraction (SAED). The HRTEM images confirmed the presence of triangle, radial, hexagonal, rod, and rectangle, shaped with an average size of 29 ± 1.3 nm. The functional groups for synthesized ZnO NPs were 3852 cm−1 for H-H weak peak, 3138 cm−1 for aromatic C-H extend, and 1648 cm−1 for Aromatic ring stretch. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), caspase and DNA fragmentation assays were carried out using various concentrations of ZnO NPs ranging from 1 to 100 mg/mL. The synthesized ZnO NPs showed dose dependent cytopathic effects in the Hep-G2 cell line. At 100 mg/mL concentration, the synthesized ZnO NPs exhibited significant cytotoxic effects and the apoptotic features were confirmed through caspase-3 activation and DNA fragmentation assays.

In recent years, there has been a significant focus on the green synthetization of metal oxide nanoparticles due to their environmentally friendly features and cost-effectiveness. The aim of this study is to biosynthesize zinc oxide nanoparticles (ZnO NPs) through a green method, utilizing crude banana peel extract as reducing and capping agents, to characterize the synthesized ZnO NPs and test their antibacterial activity. ZnO NPs were biosynthesized using the peel extract of banana with various concentrations of zinc acetate dihydrate salt, followed by annealing at 400 °C for 2 h. The synthesized ZnO NPs were characterized using UV–visible spectroscopy (UV-Vis), scanning electron microscopy (SEM), dynamic light scattering (DLS), attenuated total reflectance–Fourier-transform infrared (ATR-FTIR), and X-ray diffraction (XRD). Also, its antibacterial efficiency against different bacterial strains was tested. ZnO NPs were biosynthesized successfully using the extract of Musa Acumniata (cavendish) fruit peel with a UV-Vis wavelength range of 344 to 369 nm and an electrical band gap ranging from 3.36 to 3.61 eV. The size varied from 27 ± 4 nm to 89 ± 22, and the negative zeta potential (ζ) ranged from −14.72 ± 0.77 to −7.43 ± 0.35 mV. ATR-FTIR analysis showed that the extract phytochemical functional groups were present on ZnO NPs. XRD results confirm the formation of a highly pure wurtzite hexagonal structure of ZnO NPs. Moreover, the best obtained size of ZnO NPs was selected for the antibacterial tests, giving the highest inhibition growth rate against Staphylococcus epidermidis (98.6 ± 0.9%), while the lowest rate was against Pseudomonas aeruginosa (88.4 ± 4.4%). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were reported and compared to previous studies. The unique properties of greenly synthesized ZnO NPs and their antibacterial activity have potential for reducing environmental pollution and the use of antibiotics, which may contribute to solving the problem of bacterial resistance. Therefore, studies that aim to design an applicable dosage form loaded with biosynthesized ZnO NPs might be conducted in the future.

In vitro cytotoxicity of silver nanoparticles and zinc oxide nanoparticles to human epithelial colorectal adenocarcinoma (Caco-2) cells

Fungicides have been extensively used to control fungal diseases that affect several crops including ornamental crops. However, concerns have arisen due to a development of fungicide resistance and increasing incidences of fungicide toxicity effects on nontarget organisms. As zinc oxide nanoparticles (ZnO NPs) have demonstrated effective antimicrobial activity, this study is therefore aimed at synthesizing ZnO NPs from banana peels using a green chemistry method in a large scale and determines their physical properties including their inhibitory effects against a plant pathogen fungus causing anthracnose in orchids, Colletotrichum sp. Results from X-ray diffraction and scanning electron microscope indicated that the synthesized ZnO NPs were obtained without other crystalline impurities, and they were spherical in shape with the average diameter of 256 ± 40 nm , respectively. The absorption peak was found to be centered at ~370 nm with the optical band gap value approximately 2.8 eV. Fourier transform infrared spectroscopy analysis confirmed the presence of several functional groups on synthesized ZnO NPs. The total amount of synthesized ZnO NPs was obtained about 170 g for a synthesis reaction. By performing the antifungal activity assay, high doses of green synthesized ZnO NPs significantly inhibited growth of isolated Colletotrichum sp. (KUFC 021) on culture plates. Under greenhouse conditions, high doses of synthesized ZnO NPs also significantly reduced anthracnose symptoms on inoculated orchid leaves with the Colletotrichum sp. (KUFC 021).

This study aimed to investigate the characterization of zinc oxide nanoparticles (ZnONPs) produced from Cucurbita pepo L. (pumpkin seeds) and their selective cytotoxic effectiveness on human colon cancer cells (HCT 116) and African Green Monkey Kidney, Vero cells. The study also investigated the antioxidant activity of ZnONPs. The study also examined ZnONPs' antioxidant properties. This was motivated by the limited research on the comparative cytotoxic effects of ZnO NPs on normal and HCT116 cells. The ZnO NPs were characterized using Fourier-transform infrared spectroscopy (FTIR), Thermogravimetric Analysis (TGA), Transmission Electron Microscope/Selected Area Electron Diffraction (TEM/SAED), and Scanning Electron Microscope-Energy Dispersive X-ray (SEM-EDX) for determination of chemical fingerprinting, heat stability, size, and morphology of the elements, respectively. Based on the results, ZnO NPs from pumpkins were found to be less than 5 μm and agglomerates in nature. Furthermore, the ZnO NPs fingerprinting and SEM-EDX element analysis were similar to previous literature, suggesting the sample was proven as ZnO NPs. The ZnO NPs also stable at a temperature of 380°C indicating that the green material is quite robust at 60-400°C. The cell viability of Vero cells and HCT 116 cell line were measured at two different time points (24 and 48 h) to assess the cytotoxicity effects of ZnO NP on these cells using AlamarBlue assay. Cytotoxic results have shown that ZnO NPs did not inhibit Vero cells but were slightly toxic to cancer cells, with a dose-response curve IC50 = ~409.7 μg/mL. This green synthesis of ZnO NPs was found to be non-toxic to normal cells but has a slight cytotoxicity effect on HCT 116 cells. A theoretical study used molecular docking to investigate nanoparticle interaction with cyclin-dependent kinase 2 (CDK2), exploring its mechanism in inhibiting CDK2's role in cancer. Further study should be carried out to determine suitable concentrations for cytotoxicity studies. Additionally, DPPH has a significant antioxidant capacity, with an IC50 of 142.857 μg/mL. RESEARCH HIGHLIGHTS: Pumpkin seed extracts facilitated a rapid, high-yielding, and environmentally friendly synthesis of ZnO nanoparticles. Spectrophotometric analysis was used to investigate the optical properties, scalability, size, shape, dispersity, and stability of ZnO NPs. The cytotoxicity of ZnO NPs on Vero and HCT 116 cells was assessed, showing no inhibition of Vero cells and cytotoxicity of cancer cells. The DPPH assay was also used to investigate the antioxidant potential of biogenic nanoparticles. A molecular docking study was performed to investigate the interaction of ZnO NPs with CDK2 and to explore the mechanism by which they inhibit CDK2's role in cancer.

Development of novel green synthesized Zinc oxide nanoparticles with antibacterial activity and effect on diabetic wound healing process of excisional skin wounds in nursing care during sports training

The aim of the present study was to ascertain whether or not autophagy is induced by tanshinone IIA (TanIIA), and to explore the crosstalk between autophagy and apoptosis in regards to the antitumor effects of TanIIA on MG-63 cells and the potential mechanism. MG-63 cells were cultured in vitro with various concentrations of TanIIA (0, 2.5, 5, 10 and 20 mg/l) for 0, 24, 48 and 72 h, respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay was used to evaluate the inhibition of the proliferation of osteosarcoma MG-63 cells by TanIIA or in the presence/absence of chloroquine (CQ). Autophagic vacuoles and characteristic autophagosomes were observed by transmission electron microscopy (TEM). TanIIA-induced autophagy in MG-63 cells was confirmed by GFP-LC3 punctate fluorescence. The expression levels of apoptosis-related proteins caspase-3, caspase-8, caspase-9 and cleaved-PARP and autophagy-related proteins LC3II/LC3I and Beclin-1 were detected by western blotting. FITC-Annexin V/propidium iodide (PI) staining, flow cytometry and Hoechst 33258 staining were used to analyze the apoptotic rate. Fluorescence intensity of reactive oxygen species (ROS) was examined under a fluorescence microscope using an analysis software system. Cell proliferation was obviously inhibited by TanIIA in a dose- and time-dependent manner. Generation of autophagy was triggered by TanIIA (0–20 mg/l) treatment, and in a Beclin-1-dependent manner. Compared with the control group, the apoptosis ratio following treatment with 2.5 mg/l TanIIA failed to achieve statistical significance. Expression of caspase-3, -8 and -9, and cleaved-PARP in the other groups was gradually enhanced in dose-dependent manner. Our analysis also suggested that the influence of autophagy on TanIIA cytotoxicity had a phase effect; with low-dose drugs and shorter treatment periods, autophagy functioned as a damage repair mechanism. In conrast, when the cells were treated with higher doses of TanIIA for longer treatment periods, autophagic cell death contributed to apoptosis. Furthermore, generation of ROS occurred in a dose-dependent manner and pretreatment with NAC, a selective ROS scavenger, blocked the coexistence of Beclin-1 autophagy and caspase-dependent apoptosis. In conclusion, our findings provide strong evidence that TanIIA may be a potential therapeutic drug against osteosarcoma. Moreover, its cytotoxity can be enhanced with ROS agonists.

This study explains the biosynthesis, characterization, evaluation of the antibacterial activity and cytotoxicity of zinc oxide nanoparticles (ZnO NPs) prepared by a low-cost and simple procedure. Bio-inspired ZnO NPs were synthesized with the aid of a novel, non-toxic, ecofriendly biological material namely; Banana Peels Extract (BPE). Qualitative phytochemical screening of the aqueous fruit peels extract of banana revealed the presence of many phytocomponents in it. The structural, morphological and optical properties of the synthesized nanoparticles have been characterized by using UV-Vis spectrophotometer, XRD, FE-SEM with EDX analysis, AFM and FTIR. The synthesized ZnO NPs were characterized by a peak at 373 nm in the UV-Vis spectrum. The XRD of the sample revealed the hexagonal wurtzite structure with an average grain size 11.98 nm. Particle shapes and sizes were determined by FE-SEM. Surface morphology of the sample was studied by AFM. The FT-IR confirmed the presence of functional groups of both leaf extract and ZnO NPs. The results showed the antibacterial activity of the ZnO NPs against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli). The cytotoxicity of ZnO NPs was tested against vero101 normal cell line and skin A431 cancer cell line by Crystal violet (CV) assay. Banana peels mediated ZnO NPs showed no evidence of toxicity against normal cell line, the nanoparticles are biocompatible and are non-toxic and effective cytotoxic effect against cancer cell line. These results clearly support the benefits of using a biological method for synthesizing ZnO NPs with anticancer activities.

The effects of three preservation methods (ultrasound, ZnO NPs, and ultrasound combined with ZnO NPs) on the odor, microstructure, and edible quality of fresh-cut lettuce were investigated in this study. When stored for 8 days, significant improvements were observed in the following when using ultrasound combined with ZnO NP treatment to better preserve fresh-cut lettuce (and were reduced when compared with the control group): the color (L* value (34.53); a* value (−5.89); b* value (15.00); browning index (40.63); firmness (25.66); propectin (2.12%); chlorophyll (2.75 mg/100 g); cellulose (20.35%); total phenolic content (0.95 mg/100 g); PAL activity (54.91 U·h−1·g−1); CAT activity (41.78 U·min−1·mg−1); ABTS free-radical scavenging ability (137.62 µmol/L); FRAP total reducing ability (94.42 µmol/L) (p < 0.05), PPO activity (0.85 U·min−1·g−1); MDA (1.97 µmol/g); and H2O2 (54.26 µmol/g). In addition, the results of the volatile components indicated that the use of ultrasound combined with ZnO NP treatment decreased the production of adverse flavor compounds by inhibiting the generation of aldehydes and ketones, as well as by promoting the generation of olefins, nitriles, and quinolines, and the contents of nitriles and quinolines were 20.07% and 2.07% of the total components, respectively. The resultant microstructure indicated that the microchannels generated by ultrasound allowed for the ZnO NPs to enter the intracellular cavity of the fresh-cut lettuce more efficiently; such a finding could serve as a basis for a hypothesis on the mechanism of ultrasound combined with ZnO NP treatment. The results of fresh-cut lettuce preservation when using ultrasound combined with ZnO NPs were better than those that were obtained when using ultrasound and ZnO NP treatment alone. And, using ultrasound combined with ZnO NP treatment as a new preservation method for fresh-cut lettuce provides a promising preservation idea for other fresh-cut fruits and vegetables.

Simple SummaryThe overuse of antibiotics in the poultry industry has led to the emergence of multidrug-resistant microorganisms. Thus, there is a need to find an alternative to conventional antibiotics. Recently, zinc oxide nanoparticles (ZnO NPs) have gained much attention due to their excellent antibacterial activity. In addition, ZnO NPs is an essential trace mineral in poultry diets. In this sense, incorporating ZnO NPs into poultry can promote growth and performance while serving as an alternative antibacterial agent to control diseases. Therefore, this study aimed to assess the in vitro antibacterial activity and antibacterial mechanisms of ZnO NPs against poultry-associated foodborne pathogens (Salmonella spp., Escherichia coli, and Staphylococcus aureus). The obtained findings demonstrated effective antibacterial actions against the tested microorganisms. The nanotechnology approach could represent a new tool for combating pathogens in the poultry industry.Since the emergence of multidrug-resistant bacteria in the poultry industry is currently a serious threat, there is an urgent need to develop a more efficient and alternative antibacterial substance. Zinc oxide nanoparticles (ZnO NPs) have exhibited antibacterial efficacy against a wide range of microorganisms. Although the in vitro antibacterial activity of ZnO NPs has been studied, little is known about the antibacterial mechanisms of ZnO NPs against poultry-associated foodborne pathogens. In the present study, ZnO NPs were successfully synthesized using Lactobacillus plantarum TA4, characterized, and their antibacterial potential against common avian pathogens (Salmonella spp., Escherichia coli, and Staphylococcus aureus) was investigated. Confirmation of ZnO NPs by UV-Visual spectroscopy showed an absorption band center at 360 nm. Morphologically, the synthesized ZnO NPs were oval with an average particle size of 29.7 nm. Based on the dissolution study of Zn2+, ZnO NPs released more ions than their bulk counterparts. Results from the agar well diffusion assay indicated that ZnO NPs effectively inhibited the growth of the three poultry-associated foodborne pathogens. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were assessed using various concentrations of ZnO NPs, which resulted in excellent antibacterial activity as compared to their bulkier counterparts. S. aureus was more susceptible to ZnO NPs compared to the other tested bacteria. Furthermore, the ZnO NPs demonstrated substantial biofilm inhibition and eradication. The formation of reactive oxygen species (ROS) and cellular material leakage was quantified to determine the underlying antibacterial mechanisms, whereas a scanning electron microscope (SEM) was used to examine the morphological changes of tested bacteria treated with ZnO NPs. The findings suggested that ROS-induced oxidative stress caused membrane damage and bacterial cell death. Overall, the results demonstrated that ZnO NPs could be developed as an alternative antibiotic in poultry production and revealed new possibilities in combating pathogenic microorganisms.

The purposes of this work are to evaluate the antimicrobial, antibiofilm, anticancer, and antioxidant abilities of anisotropic zinc oxide nanoparticles (ZnO NPs) synthesized by a cost-effective and eco-friendly sol-gel method. The synthesized ZnO NPs were entirely characterized by UV-Vis, XRD, FTIR, HRTEM, zeta potential, SEM mapping, BET surface analyzer, and EDX elemental analysis. Antimicrobial and antibiofilm activities of ZnO NPs were investigated against multidrug-resistant (MDR) bacteria and yeast causing serious diseases like urinary tract infection (UTI). The anticancer activity was performed against Ehrlich ascites carcinoma (EAC). Additionally, antioxidant scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) was observed. The synthesized ZnO NPs exhibited an absorption peak at 385.0nm characteristic to the surface plasmon resonance (SPR). Data obtained from HRTEM, SEM, and XRD confirmed the anisotropic crystalline nature of the prepared ZnO NPs with an average particle size of 68.2nm. The calculated surface area of the prepared ZnO NPs was 10.62m2/g and the porosity was 13.16%, while pore volume was calculated to be 0.013cm3/g and the average pore size was about 3.10nm. The prepared ZnO NPs showed promising antimicrobial activity against all tested UTI-causing pathogens. It showed a prominent antimicrobial capability against Candida tropicalis with a zone of inhibition (ZOI) reaching 22.4mm, 13mm ZOI for Bacillus subtilis, and 12.5mm ZOI for Pseudomonas aeruginosa. Additionally, the prepared ZnO NPs showed enhanced biofilm repression of about 79.33%, 72.94%, and 33.68% against B. subtilis, C. tropicalis, and P. aeruginosa, respectively. Moreover, the prepared ZnO NPs had a powerful antioxidant property with 33.0% scavenging ability after applied DPPH assay. Surprisingly, upon ZnO NPs treatment, cancer cell viability reduced from 100 to 58.5% after only 24h due to their unique antitumor activity. Therefore, according to these outstanding properties, this study could give insights for solving serious industrial, pharmaceutical, and medical challenges, particularly in the EAC and UTI medications.