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. 2025 Jan 2;15:1. doi: 10.1186/s13568-024-01808-y

Bimetallic selenium/zinc oxide nanoparticles: biological activity and plant biostimulant properties

Samy Selim 1,✉, Amna A Saddiq 2, Ruba Abdulrahman Ashy 2, Afra Mohammed Baghdadi 2, Ashwag Jaman Alzahrani 2, Ehab M Mostafa 3, Soad K Al Jaouni 4, Mohammed Yagoub Mohammed Elamir 1, Mohamed A Amin 5,✉, Ahmed M Salah 6, Nashwa Hagagy 7,8
PMCID: PMC11695524  PMID: 39747711

Abstract

Extracts of medicinal seeds can be used to synthesize nanoparticles (NPs) in more environmentally friendly ways than physical or chemical ways. For the first time, aqueous extract from unexploited grape seeds was used in this study to create Se/ZnO NPS utilizing a green technique, and their antimicrobial activity, cytotoxicity, antioxidant activities, and plant bio stimulant properties of the economic Vicia faba L. plant were evaluated. Se/ZnO NPS is characterized by SEM and TEM images, FTIR, and XRD. Through the well diffusion assay and the scavenging of 1,1-Diphenyl-2-picrylhydrazyl (DPPH) free radical experiment, biogenic Se/ZnO NPs demonstrated their antibacterial and antioxidant activities. The nanomaterial compound showed the highest inhibitory effects of 99.7, 55.63, 16.91, 10.25, 6.61, 3.83, 3.00, and 2.59%, respectively, against the cervical carcinoma (SKOV3 cells) cell line at conc of 7.81, 15.62, 31.25, 62.5, 125, 250, 500, and 1000 µg/ml, respectively, with IC50 values at 20.31 µg, resulting in 50% cell death. This study demonstrated the value of bimetallic nano-fertilizers Se/Zn in promoting faba bean development, yield features, and metabolite contents (protein, phenol, carbs, and pigments). These fertilizers are probably also advantageous for other crops. When applied in contrast to the control, 100 ppm of biological nano-Se/ZnO may generally result in the best growth and yield of faba beans. Further research is needed on the ecological aspect of biological nanofertilizers in addition to the economic one.

Supplementary Information

The online version contains supplementary material available at 10.1186/s13568-024-01808-y.

Keywords: Grape seeds, Cervical carcinoma, Multidrug resistance bacteria, Vicia faba

Key points

  1. Aqueous extract from grape seeds used a green technique in create Se/ZnO NPs.

  2. Se/ZnO NPs appeared inhibitory effects against the cervical carcinoma (SKOV3 cells) cell line with IC50 values at 20.31 µg and inhibition the growth of m multidrug resistance bacteria as S. aureus.

  3. Se/ZnO NPs had successfully scavenged the DPPH free radicals, with an IC50 value of 14.92 µg/ml and promotion growth and yield of faba bean plants.

Introduction

A new trend in the development of nanotechnology that is safer and more ecologically friendly is the production of green nanoparticles (NPs). Biological agents include fungi, bacteria, and plant extracts. Utilized in the green synthesis process as an alternative to hazardous chemicals (Ahmed et al. 2016; El-Batal et al. 2023; Amin et al. 2024; Qanash et al. 2024). The most useful and promising nanostructures for biological applications are metal NPs, these particles are distinct due to their physical, chemical, and biological characteristics (Adeyemi and Sulaiman 2015; Abdelhady et al. 2024).

In our study biogenic Se/Zn oxide NPs were obtained from grape seed extract, there are several forms of dietary supplemental grape seed extract (GSE) including powder, drink, pill, and tablet (Nassiri-Asl and Hosseinzadeh 2016. Surprisingly, the majority of the polyphenols in the fruit are found in the seeds of grapes. GSE is composed of a range of vitamins, 3% minerals, 35% fibers, 13% lipids, 11% proteins, and 7% water. It also contains a heterogeneous mixture of polyphenolic compounds, comprising 11–39% polymers, 17–63% oligomers, and 5–30% monomers (Ma and Zhang 2017.

Since zinc is required for more than 300 enzymes and hormones, it is a vital micronutrient for both humans and plants. A variety of metabolic activities involve zinc, like protein synthesis, carbohydrate metabolism, DNA transcription, RNA processing, and cell membrane integrity. In plant, zinc deficiency in plants is associated with reduced rate of photosynthesis, stunted development, hormone abnormalities, chlorosis, spikelet volume infertility, poorer crop yield, and lower nutritional content in fruits and seeds (El-Ramady et al. 2022). Also, living things needs selenium (Se), a trace element. Due to Bio-Se NPs’ great degradability, low toxicity, and capacity to be eliminated by the body, they are also relatively new (Abbas and Abou Baker 2020). Biogenic Se NPS are harmless and biodegradable in comparison to other NPs. (Wadhwani et al. 2016). In a recently study on trimetallic Se/ZnO/CuO NPs prepared by extract of Nitraria retusa leaves showed promising antimicrobial, antioxidant, anticancer and antidiabetic properties (Amin et al. 2025).

With concerning NPs and the vital role on plants, Ikram et al. (2020), found that SeNPs promote organogenesis and root development. Therefore, seed priming with ZnONPs reduces ROS damage in rapeseed and positively influences growth characteristics under salt stress (El-Badri et al. 2021.

Being a major crop in Northern Africa and a rich source of minerals and organic materials (proteins, carbohydrates, thiamin, folic acid, and tocopherols), faba bean, is utilized in the food business and is regarded as one of the most important sources of nourishment for both humans and animals, Moreover, faba bean plants improve soil fertility and production by adding nitrogenous molecules to the soil (Qados et al. 2015).

The purpose of this study is to determine the production of Bio-Se/ZnO NPs by using active ingredients of grape seeds extract as a natural precursor in this process and their effects as antimicrobial agents. Cytotoxicity, antioxidant and antitumor, antimicrobial activities also have been assessed. Also, the goal of our research is to examine how Se/ZnO NPs affect the vegetative growth, yield traits and various physiological traits as photosynthetic pigments, carbohydrates, protein, and phenol accumulation of faba bean plants. This is done by using natural chemicals that are delivered into plant tissues through NPs, which may promote the growth and metabolites of plants. To the best of our knowledge, this is the initial research study to be published on the topic of biological Se/ZnO NPs involvement in promoting economic faba bean plant growth.

Materials and methods

Aqueous seed grape extraction

The grape seeds (Egyptian Queen Nina grapes) were cleaned with distilled water, allowed to air dried for two weeks, and then ground into powder. In order to create the watery extract of grape seeds, 5 g of air-dried powder were added to 100 ml of DW and sonicated for two hours at 50 °C in a water bath. Filter paper (Whatman No. 1) was then used to filter the extract, and the filtrate was then kept in a refrigerator at 4 °C.

Biosynthesis of Se/Zn NPs

Se/Zn NPs were produced by adding 30 mL of an aqueous grape seed extract to a 250-milliliter flask and stirring with a magnetic stirrer. Following that, 10 ml of sodium selenite (1 mM) and 10 ml of zinc nitrate (1 mM) were added to the flask. The mixture was stirred for 48 h at 25 °C. The hue shift of the mixture confirmed that Se/Zn NPs were produced. The produced NPs were precipitated by centrifugation at 14,000 rpm for 20 min at 4 °C. Before being freeze-dried, the pellets were thoroughly cleaned in DW water (Mirzaei et al. 2021).

Characterization of Se/Zn NPs

An X-ray diffractometer (D8-Advance; Bruker, Germany) was utilized to evaluate the microstructure of the nanomaterial. The morphology of the generated samples was examined using transmission electron microscopy (TEM) (JEM-6700 F; JEOL, Tokyo, Japan). TEM was utilized to evaluate the size and shape of the generated NPs. X-ray (EDX) spectrometer, the composition of nanomaterial was examined. The molecular vibrations of the generated samples were investigated using Fourier transform infrared spectroscopy. FTIR, is a technique that may identify a range of functional groups by using the absorption of infrared radiation from the sample’s substance, which can be solid, liquid, or gas.

Detection of anti-bacterial activity

Selinium/Zinc oxide NPS made from grape seed extract and doped with selenium were studied for their antimicrobial qualities using the diffusion method by using an agar well. This study used potato dextrose agar medium for fungi and Mueller-Hinton agar (MHA) media for bacteria to cultivate a variety of microorganisms. Salmonella typhi (ATCC 6539), Bacillus cereus (ATCC 11778), Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 8739), Candida albicans (ATCC 10221), and Aspergillus niger (ATCC 16888) were among these microbes. Erythromycin/Nystatin for bacteria/fungi serves as the positive control. 150 µL of selenium-doped ZnO NPs were put one at a time to every petri plate well. Following a one day incubation period at 37 °C, the millimeter-scale zones of inhibition on the plates were assessed (Abdelghany et al. 2023).

Antioxidant activity

Se/ZnO NPs’ DPPH scavenging ability was evaluated. While the DPPH was dissolved in methanol, various double-fold doses of the investigated substances, from 1000 to 1.95 µg/ mL, were dissolved in high pure H2O (Milli Q H2O). 1.0 mL of each concentration, 1.0 mL of DPPH, and 450 µL of Tris-HCl buffer (pH = 7.4) were combined in a test tube. Each tube’s mixture was thoroughly mixed before being shaken (at 150 rpm) and incubated in darkness for 1/2 hour at 37°C. Ascorbic acid was utilized as a positive control at the same quantities, while a test tube with every component absent but the compounds being tested (ascorbic acid or NPs) served as the negative control. The scavenging action was identified at 517 nm, and the percentage was computed as follows (Alawlaqi et al. 2023):

graphic file with name M1.gif

In-vitro cytotoxicity

Carcinoma cell (i.e., the cervical cancer cell line, SKOV3 cells) was used in the viability experiment for the nanomaterial’s antitumor test utilizing the MTT assay method. The source of these cells was the Biological Products and Vaccines Company (VACSERA), located in Cairo, Egypt.

MTT protocol

The chosen normal cell grown with 96 wells in a culture plate and incubated until the intensity reached 1 × 105 cells per 100 µL per well. After that, the cells were moved to a CO2 incubator (5%) that was regulated at 37 °C for a day. Following the formation of a monolayer sheet, 100 µL of RPIM media was added. Next, treatments with plant extract and nanoalloy at varying concentrations (1000–31.25 µg/mL) were applied, and 48 h of 37 °C incubation were spent on the combination. Three wells treated with DMSO (solvent system) served as the control. After removing excess medium, the cells were treated with MTT solution (50 µL, 5 mg mL–1) at the end of 48 h. They were then thoroughly mixed and incubated for five hours at 37 °C. The formazan crystal that had formed as a result of MTT metabolism was then dissolved by treating each well with 100 µL of 10% DMSO after the excess solution from the MTT reagent had been removed. After thirty minutes, the DMSO was taken out and 570 nm color absorbance was measured. The following formula (Selim et al. 2024) was used to get the cell viability percentages.:

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A pot experiment showing the effect of selenium/zinc oxide NPs on morphological and biochemical traits of Vicia faba L

The seeds of Vicia faba L. variety Giza 22 (faba bean) were sourced from Agricultural Research Centre, Egypt. 30 cm-diameter pots with 6.0 kg of loamy soil used to sowing Vicia faba seeds. Four groups of pots were created, each of which represented a distinct kind of treatment. (1) Tape water (Control); (2) 50 ppm NPS (soaking seeds 6 h) (3) 100 ppm NPS (soaking seeds 6 h) (4) 200 ppm NPS (soaking seeds 6 h) At 45 days next planting and the end-of-life cycle of plant, samples from the different treatments and the control were taken for estimated growth traits, pigments contents, carbohydrate, protein, and phenol contents.

Pigment and carotenoid assessment

one gram of fresh leaves was extracted by 100 ml of 80% acetone. The quantities of chlorophylls (a and b), as well as their total amounts in plant tissues by Vernon and Selly (1996). For the estimation of carotenoid chemical composition, the method described by Lichtenthaler et al. (1981).

Quantification of soluble carbohydrates and protein

To extract soluble carbohydrates, five ml of 2% phenol water and 10 mL of 30% trichloroacetic acid added to one gram of plant powder. it was left to sit for the entire night before filtering. Following that, the volume of the filtrate was lowered to 50 mL (Said et al. 1964). The anthrone technique for quantifying soluble carbohydrates (Umbreit et al. 1957). To extract soluble protein, one gram of the finely ground added 10 mL of distilled water and combined with 5 mL of 2% phenol water. Following filtration of the reaction mixture, the filtrate’s ultimate volume of 50 ml was adjusted using distilled water.

The contents of proteins were ascertained according to (Lowry et al. 1951).

Estimation of total phenolic compounds

According to (Cumplido-Nájera et al. 2019), phenols (mg 100 g\1 DW) were assessment by using Folin-Ciocalteu reagent. A test tube containing fifty mL of 20% Na2CO3, fifty mL of distilled water, two mL of the Folin-Ciocalteu reagent, and fifty mL of the phenol extract was vortexed for thirty seconds. Ultimately, the reading at 750 nm of absorbance was obtained using a plastic photocell in a UV-vis spectrophotometer.

Analytical statistics

To do statistical computations at level 0.05 of probability, the computer applications Minitab version 19, Microsoft Excel version 365, and Minitab 18 were employed. Analyses of variance, one-way ANOVA, and post hoc Tukey’s test were employed to examine parametrically distributed quantitative data.

Results

Characterization of Se/ZnO NPs (TEM, SEM and XRD)

Dimensions and shape of the produced nanomaterial were examined using a TEM, in Fig. 1A. The sample’s TEM picture, displays typical spherical NPs. their sizes which ranged from 7.67 to 12.86 nm were ascertained. The morphology, surface topology, and pore size of the produced nanoparticle material are visible under a scanning electron microscope. The SEM picture of Se/Zn NPs with a smooth, spherical surface is shown in Fig. 1B, Se/Zn NPs seem to have a multifaceted structure and a homogeneous size. This corresponds to the planes that its hexagonal structure, as determined by XRD, forms. Significant roughening of the Se/Zn nanoparticle surfaces (Fig. 1C). The most efficient technique for figuring out the phase and crystallite size of the prepared Se/Zn NPs is to use the X-ray diffraction pattern. Particle sizes are in the nanoscale range, as the images shows. The XRD peaks show that the synthesized Se/Zn NPs’ particles were crystalline in nature, which is consistent with the data from the JCPDS (File No. 36-1451) for ZnO NPs and the JCPDS (File No. 06-0362) for SeNPs. The XRD patterns’ expanding peaks show that tiny Se/Zn nanocrystals are forming. When creating NPs, the broadening peaks are important. Conversely, a decrease in size is indicated by the decreased intensity. Debye Scherer’s formula (D = Kλ / βCosθ) is used to determine that Se doped ZnO NPs have a normal size of 27 nm. The SAED pattern of the produced SeNPs showed that they resembled crystals (Fig. 1D).

Fig. 1.

Fig. 1

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Characterization of bio- synthesized Se/ZnO NPS. (A) Shows TEM image, (B) SEM image, (C) XRD spectrum and (D) SAED pattern

FT-IR Spectra

The functional groups found in the active substances in the extract and manufactured NPs were identified using the FT-IR spectra. The FT-IR spectrum data confirmed the existence of N-H, O-H, C = C, C-H, C = O, CH2, C-O, and CH3 functional groups. The stretching vibration of the overlapping N-H and O-H was identified as the cause of the appearance peak at 3243 cm− 1. The presence of polysaccharides was attributed to the apparent peak at 1592 cm− 1. Amino acids exhibit COO− stretching at 1592 cm− 1 and N-H bending at 1495 cm− 1. The methyl group that bends has an absorption band at 1372 cm− 1. Peak at 1110 cm− 1 is attributed to the overlapping stretching vibrations of C-NH and C-OH. The aromatic group’s C-H bending vibration was attributed to the peak at 720. C-Br or C-I vibration could be responsible for the peak at 616 cm− 1.The SeNPs scaffold had a distinct spectrum. Also, in Fig. 2. As shown, the spectra of Se-ZnO -NPs synthesis show appearance of new peaks or alter the intensity of the existing peaks. The new absorption band appeared at 3284 cm− 1 was assigned to overlapping O-H and N-H stretching vibration (Ruan et al. 2023). The C-H aliphatic streatching of methyl group was appeared at 2923 cm− 1 while the C-H stretching aliphatic of methylene was appeared at 2853 cm− 1. The methyl group’s C-H bending vibration was observed at 1372, whereas the methylene group’s C-H bending vibration was observed at 1441 cm− 1.The aromatic ring’s C-H stretching was attributed to the absorption band that occurred at 3009 cm − 1. While the aromatic ring’s C-H bending vibration was responsible for the peak that occurred at 762 cm− 1. Strong absorption band appeared at 1743 cm− 1 was assigned to C = O stretching vibration. Ring stretch absorption often occurred in pairs at 1605 cm− 1 and 1518 cm− 1. Also, amino acids show N-H bending at 1518 cm− 1 and COO− stretching at 1605 cm− 1. EDX is an analytical technique that offers both type and percentage the elemental structure of a prepared sample. The elemental structure of Se/ZnO NPS portrays in Fig. 3. This figure is confirmed zinc and selenium in nanomaterial.

Fig. 2.

Fig. 2

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FT-IR analysis of plant aqueous extract and bimetallic Se/ZnO NPS showing different functional groups

Fig. 3.

Fig. 3

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EDX analysis of green-fabricated Se/ZnO NPS showing their metallic components

Evaluation of the antitumor activities

The antitumor test of the nanomaterial was performed using viability assay on one carcinoma cell (i.e. cervical cancer cell line (SKOV3 cells). As shown in Table 1; Fig. 4. The nanomaterial compound showed the highest inhibitory effects of 99.7, 55.63, 16.91, 10.25, 6.61, 3.83, 3.00 and 2.59%, respectively, against Cervical carcinoma (SKOV3 cells) cell line at conc of 7.81, 15.62, 31.25, 62.5, 125, 250, 500 and 1000 µg/ml, respectively. In our study, the nanomaterial compound showed inhibitor effects against Cervical carcinoma (SKOV3 cells) cell line with IC 50 at 20.31 µg/ml.

Table 1.

The viability and toxicity percentage of Se/Zn NPS against cervical cancer cell line (SKOV3 cells)

concentration (µg/ml) 1000 500 250 125 62.5 31.25 15.62 7.81 0
Viability % 2.59 3.00 3.83 6.61 10.25 16.91 55.63 99.77 100
Toxicity % 97.40 96.99 96.16 93.38 89.74 83.08 44.36 0.22 0
IC 50 20.31
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Fig. 4.

Fig. 4

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Shows the effect Se/ZnO NPS on SKOV3 cells at different concentration

Evaluation of antioxidant activities

In Table 2, demonstrates the antioxidant activity of Se/Zn NPs, which increased DPPH scavenging as concentration increased. Using Se/Zn NPs, the DPPH scavenging percentage was 24.9 ± 1.90, 62.1 ± 1.67, 81.7 ± 1.32 and 90.4 ± 3.45% at 1.95, 62.50, 250 and 500 µg/ml respectively. The DPPH assay findings showed that Se/ZnO NPs had successfully scavenged the DPPH free radicals, with an IC50 value of 14.92 µg/ml, in comparison to the standard (ascorbic acid) of 4.52 µg/ml.

Table 2.

Antioxidative activity of the nanomaterial compound at 1 mg/ml by DPPH free radical scavenging assay

Concentration (µg/mL) DPPH scavenging% HSD
Se/Zn NPs Ascorbic acid
1000 94.9 ± 1.23 97.3 ± 0.87 4.68
500 90.4 ± 3.45 92.3 ± 1.25 6.27
250 81.7 ± 1.32 91.8 ± 2.92 5.94
125 73.0 ± 3.45 82.9 ± 3.65 8.38
62.50 62.1 ± 1.67 76.2 ± 1.50 11.35
31.25 56.8 ± 0.86 65.1 ± 0.67 9.247
15.63 46.5 ± 2.76 58.6 ± 0.54 10.25
7.81 39.6 ± 1.34 53.4 ± 2.54 8.81
3.90 29.5 ± 2.89 48.1 ± 2.67 12.32
1.95 24.9 ± 1.90 39.7 ± 1.56 7.25
0.0 0.0 0.0 00
IC50 14.92 µg/ml 4.52 µg/ml 7.21
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Evaluation of antibacterial activities

As mentioned in Table 3 and visualized in Fig. 5, the applied Se/ZnO NPs reflected excellent antibacterial activity besides anticandidal activity while failed to prevent the growth of (A) niger. Higher inhibition zones 29 ± 0.2, 28 ± 0.2, 50 ± 0.3, and 32 ± 0.2 mm was attributed to Se/ZnO NPs than that at standard drug 25 ± 0.2, 23 ± 0.2, 30 ± 0.2, and 27 ± 0.2 mm against E. coli, S. typhi, (B) cereus and (C) albicans, respectively. Only S. aureus was more affected by standard drug than Se/ZnO NPs.

Table 3.

Antimicrobial activity of extract, ZnO@MgO NPs, Gentamycin/Nystatin as positive control, and DMSO as negative control against different microorganisms

Tested microorganisms Inhibition zones (mm)
Se/ZnO NPs Positive control (Erythromycin/Nystatin) Negative control
S. aureus 21 ± 0.1 24 ± 0.1 0.0 ± 0.0
E.coli 29 ± 0.2 25 ± 0.2 0.0 ± 0.0
S. typhi 28 ± 0.2 23 ± 0.2 0.0 ± 0.0
B. cereus 50 ± 0.3 30 ± 0.2 0.0 ± 0.0
C. albicans 32 ± 0.2 27 ± 0.2 0.0 ± 0.0
A. niger 0.0 ± 0.0 26 ± 0.2 0.0 ± 0.0
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Fig. 5.

Fig. 5

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Antimicrobial activity of ZnO@MgO NPs, Gentamycin/Nystatin as positive control (S), DMSO as negative control (NC) against different microorganisms

Growth and biochemical traits parameters in response to different concentration of Se/ZnO NPs

Agriculture and food quality are expected to be significantly impacted by the emerging field of bio-nanotechnology. Data in Tables 4 and 5, showed significant increase in growth parameter (lengths of shoot and root, fresh of shoot and root) and yield traits (Dry Weight /pod, No. of seeds/pod, Weight of 100 seeds and No. of pods/plant) when subjected to 50,100 ppm of Se/ZnO NPS, but at 200 ppm appeared drop in these parameters compared to other treatments. Se/ZnO NPS at 100 ppm showed the highest values of shoot and root lengths, fresh and dry weight of shoot by 26.14, 73.78, 51.73 and 39.75% respectively compared to control group. Regarding chlorophyll a, b, a + b and carotenoid (Fig. 1S) our results appeared the vital role of Se/Zn NPS at 50 and 100 ppm in improvement these pigments when compared to the control. Our data in Fig. 2S, demonstrated that the total soluble carbohydrates, protein and phenol contents of shoot and yield appeared increased trend with increasing Se/Zn NPs concentrations from 50 ppm up to 100 ppm. Se/Zn NPs at 200 ppm appeared significant decrease in these components. In our study Se/Zn NPs at 100 ppm appeared the highest values of carbohydrates, protein and phenol in shoot and yield.

Table 4.

Growth traits of Faba bean (shoot lengths, root lengths and fresh weight of shoot and root) in response to SeO/ZnO nanoparticles

Growth traits
Treatments Shoot lengths (cm) Root lengths (cm) Fresh weight of shoot Fresh weight of root
Control 0 ppm 44.65 ± 2.89b 6.33 ± 1.15b 19.10 ± 2.21b 3.17 ± 0.32b

SeO/ZnO

Nano.

 50 ppm 52.67 ± 3.21a 9.66 ± 0.57a 25.56 ± 4.32a 4.19 ± 0.97a
100 ppm 56.32 ± 3.06a 11 ± 1.02a 28.98 ± 3.65a 4.43 ± 0.49a
200 ppm 39.58 ± 3.12b 5.66 ± 0.44b 17.52 ± 1.21b 3.05 ± 0.21b
HSD at 0.05 7.32 2.33 5.21 0.35
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Table 5.

Yield traits of faba bean (dry weight /pod, number of seeds/pod, weight of 100 seeds and number of pods/plant) in response to different concentration of SeO/ZnO nanoparticles

Yield traits
Treatments Dry weight /pod No. of seeds/pod Weight of 100 seeds No. of pods/plant
Control 0 ppm 4.12 ± 0.081d 3.4 ± 0.41b 105.11 ± 1.21c 4.21 ± 0.32c

SeO/ZnO

Nano.

 50 ppm 6.98 ± 0.083b 5.74 ± 0.05a 112.58 ± 2.32b 5 ± 0.12b
100 ppm 8.42 ± 0.043a 6 ± 0.70a 132.78 ± a1.65a 6 ± 0.20a
200 ppm 5.58 ± 0.083c 3.8 ± 0.44b 107.54 ± 3.21c 4 ± 0.31c
HSD at 0.05 1.11 1.21 5.21 0.35
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Discussion

Innovative antibiotics and cancer treatments are now required due to rising bacterial resistance and the detrimental effects of already utilized antibacterial agents. In our study we applied grape seed as reducing agent to synthesis bimetallic NPs Se/Zn NPS and their antimicrobial activity, cytotoxicity, antioxidant activities, and plant bio stimulant properties of the economic Vicia faba L. plant were evaluated. FE-SEM was used to examine the surface morphology of the produced ZnSe NPs. The FE-SEM picture of Se/Zn NPs with a smooth, its spherical surface seems to have a multifaceted structure and a homogeneous size. The XRD peaks of our study are proposed by Dhivya et al. (2019) and Shah et al. (2010), which show that Se/Zn NPs have been dominant diffraction peaks at 2θ = 21.97, 31.854, 47.61, and 66.9720, which are assigned to the lattice plans of (110), (101), (112), and (004), respectively.

The FTIR technique was used to examine the chemical alterations on the outside of the as-synthesized nanocrystals. In FT-IR spectra a large peak in the spectra at 575 cm-1 is indicative of ZnO NPs (Shanmugam et al. 2023). The emergence of the peaks at 714. And 497 cm− 1 confirms the presence of Se (Dhivya et al. 2019). The C = O bending group that includes amines is responsible for the absorption peak at 1592.54 cm− 1, this peak was altered in their intensity, besides the appearance of new peaks at 1605 after reacting of plant extract with metal and metal oxide precursor to form Se/ZnO NPS (Ebrahimipour et al. 2014).

Two distinct peaks could be seen in the host substance of the Se/Zn NPs: one at 1.5 keV for element Se and one at 1 and for elemental Zn. Small peaks in the electromagnetic spectrum indicate the presence of minor levels of Au, which can be attributed to the coating procedure the samples underwent before performing EDX examination (Salem et al. 2017). The Zn and Se ratio is 1:1, according to EDX examinations of prepared samples.

More than 300 mammalian enzymes require zinc as a cofactor, and it is essential for the host’s defense against the development and spread of cancer. ZnO NPs, have a lot of promise for treating cancer However, the bimetallic Se/Zn NPS Not studied yet. In our study, the nanomaterial compound showed inhibitor effects against Cervical carcinoma (SKOV3 cells) cell line with IC50 at 20.31 µg/ml. In a study on ZnO-NPs appeared cell cycle arrest, promote apoptosis, and suppress SKOV3 cell growth. Moreover, ZnO-NPs prevented SKOV3 cells from invading and migrating, also, stimulated autophagy and activated endoplasmic reticulum stress (Gu et al. 2023). In a study on selenium NPs prepared by aqueous extract of Ulva fasciata indicate an effective anticancer agent against (Shahzamani et al. 2022). Tweedy’s chelation concept provided an explanation for the compound’s antitumor activity, which may be linked to the main metal atom. Additionally, the metal’s positive charge makes it more acidic, which strengthens hydrogen bonding and promotes biological activity. Furthermore, metal may promote oxidative tissue damage via a Fenton-like free-radical-mediated mechanism (Alizadeh et al. 2013). Also, metal may have two opposing effects on DNA: it may cause damage while simultaneously impeding its repair (Hegde et al. 2011). Significant volume reductions resulting from intracellular ion losses and protein loss, or from changed porosity to sodium or potassium, are indicative of damage. Bases and sugars in DNA react with OH radicals, via which both the releasing of the free base and strand breakage take place (Alizadeh et al. 2013).

Since the DPPH test is sufficiently perceptive to identify the active components at very little amounts, it is frequently employed to screen for antioxidant activity (Tettey and Shin 2019). The DPPH assay findings showed that Se/ZnO NPs had successfully scavenged the DPPH free radicals, with an IC50value of 14.92 µg/ml, in comparison to the standard (ascorbic acid) of 4.52 µg/ml. Numerous research have confirmed the antioxidant activity of zinc and selenium NPs. On the other hand, the antioxidant action of zinc doped with selenium has not been thoroughly investigated. In a recently study on ZnSe NPs algae-based synthesis found that, as ZnSe NP concentrations in the samples rose, so did the antioxidant activity. As anticipated, at 30 µg/ml, ZnSe NPs’ maximal antioxidant activity was 77.97%. At the same dose (30 µg/ml), seaweed extract demonstrated antioxidant activity of approximately 38.6 (Mirzaei et al. 2021). In one study, ZnSe NPs’ antioxidant activity reduced inflammation and promoted further cell development. According to Zheng et al. (2013), the outer charge of NPs influences both antioxidant activity and cytotoxicity. The negative-charged NPs were shown to have greater antioxidant activity, good stability, and low cytotoxicity. The outcome aligns with previous studies on single metallic NPS that employed SeNPs derived from alternative natural sources, like Spirulina platensis, which demonstrates an 89% capacity to scavenge DPPH at 500 µg/mL (Abdel-Moneim et al. 2021). According to Tettey and Shin (2019) the biologically produced ZnO NPs significantly and dose-dependently neutralized DPPH free radicals (125 µg/ml-48.78%; 250 µg/ml- 49.1%; 500 µg/ml-49.52% and 1000 µg/ml 56.11%).

The antimicrobial potential of Se/ZnO NPs was estimated against various microorganisms including fungi and bacteria in the present study. Our study reflected excellent inhibition zones ranged from 28 ± 0.2 to 50 ± 0.3 based on the tested microorganisms. Generally, metal NPs in the current decade played a vital role for inhibition the growth of pathogenic microorganisms (Abdelghany et al. 2018; Alsalamah et al. 2024) particularly bimetals of NPs (Al-Rajhi et al. 2022; Al Abboud et al. 2024; Alghonaim et al. 2024). According to Saleem et al. (2022) Enterococcus faecalis and Streptococcus mutans were inhibited by Se/ZnO NPs. At 50 µL, SeNPs repressed the growth of C. albicans with 15 ± 2.1 mm, while ZnNPs reflected inhibition zone 14 ± 2.8 mm, but when combined it provided more inhibition zone (Saleem et al. 2022). The inhibitory zone of ZnO@SeO NPs was found to be 28.33 mm against S. aureus, 27.33 mm with B. subtilis, 27.67 mm toward B. cereus, 27.17 mm on Pseudomonas aeruginosa, 27.5 mm with E. coli, 24.33 mm with S. typhi, 23.33 mm on C. albicans, and 18.17 mm on A. niger in a recent investigation when compared to undoped ZnONPs or antibiotic/antifungal. The variations in microorganism inhibitions could be attributed to the mediator of NP production.

Se/ZnO NPS 50,100 ppm appeared improvement of growth, yield traits, chlorophyll a, b, a + b, carotenoid (in leaves), total soluble carbohydrates, protein and phenol contents of shoot and yield, but at 200 ppm appeared drop in these parameters compared to other treatments. In another study at low concentration of Se or Zn NPS, all growth and yield traits of garlic plants were positively impacted by ZnO NPs (0.5, 1 and 2 ppm) and Se NPs (0.5, 1 and 2 ppm) (El-Saber 2021). Also, tomato and maize seedling vigor index was greatly enhanced (32.9–46.1%) by ZnO NPs at 75 ppm (Guardiola-Márquez et al. 2023). In another study on El-Bialy et al. (2023) demonstrated that when compared to the control, the applied 25, 50, 75, and 100 ppm of Se NPS enhanced the seedlings’ growth, pigments, antioxidant activity, and nutritional values. Foliar application of 50 ppm of Nano scale ZnO promotion the growth criteria of Vicia faba plants in comparison to control (Mogazy and Hanafy 2022. As proposed by Mahmoud et al. (2019), found that, ZnO at a rate of 60 and 50 mg/L greatly enhanced the morphological and yield traits of radish species. Furthermore, the health risks score, indicating that there was no harm to human health for radish plants that were provided with ZnO NPs. In relation to the use of ZnO-NPs, Faizan et al. (2018); found that when tomato plant roots were dipped into ZnO-NPs at concentrations of 2, 4, 8, or 16 ppm, growth and photosynthetic efficiency of the plants were considerably enhanced at 45 days after planting. Additionally, it was shown that 8 mg of ZnO-NPs L–1 was the most successful treatment. A another study on wheat and maize by using Zn NPS only (50,100,150 and 200 ppm), Srivastav et al. (2021) report that a lower dose of ZnO NPs (100 ppm) may encourage plant development and be a good source of Zn fertilizer for agricultural production.

In a study on Se-NPS, El Lateef Gharib et al. (2019), found that the biochemical measurements of the carbohydrates, protein and phenol contents in leaves of cowpea plants were improved by the application of Se-NP, especially at 6.25µM concentration. In another study on celery species, Li et al. (2020) reported that foliar treatment of 5 mg/L Se-NPs to increased total phenol content of the leaves by 21.4% in comparison with the control group. ZnO + FeO NPs at a rate of 60 and 50 ppm significantly increased the amounts of phenols, crude protein, and carbohydrates in red radish root (Mahmoud et al. 2019). The promotion role of ZnO NPs alone was observed in our earlier study (Amin and Badawy 2017), which found that Phaseolus vulgaris L. plants’ carbohydrate, protein, amylase, protease, nitrogen (N), phosphorus (P), and potassium (K) contents are all positively impacted by ZnO NPs at concentrations of 25, 50 and 100 ppm.

We are concluded that, bio-nanotechnology is a relatively new field with a lot of potential to impact agriculture and food quality. In our results demonstrates the antioxidant activity of Se/Zn NPs, which increased DPPH scavenging as concentration increased. Using Se/Zn NPs, the DPPH scavenging percentage was 24.9 ± 1.90, 62.1 ± 1.67, 81.7 ± 1.32 and 90.4 ± 3.45% at 1.95, 62.50, 250 and 500 µg/ml respectively. Also, appeared that the nanomaterial compound showed the highest inhibitory effects of 99.7, 55.63, 16.91, 10.25, 6.61, 3.83, 3.00 and 2.59%, respectively, against Cervical carcinoma (SKOV3 cells) cell line at conc of 7.81, 15.62, 31.25, 62.5, 125, 250, 500 and 1000 µg/ml, respectively. The current study found that applying Se/ZnO NPs at rates of 50 ppm and 100 ppm, enhanced the morphological, physiological, and nutritional qualities of faba bean. This study has shown that the growth, quantity, and quality of faba bean plants are all positively impacted by green synthetic nanofertilizers (Se/ZnO NPS) at 100 ppm.

Electronic supplementary material

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Supplementary Material 1. (122.4KB, docx)

Acknowledgements

The authors gratefully acknowledge the technical and financial support from Jouf University, Saudi Arabia.

Author contributions

Investigation and formal analysis S.S., R.A.A., A.M.B., and A.J.A.,; Methodology, Resources, Writing– review and editing, E.M.M.,S.K.J., M.Y.M.E., M.A.A., A.M.S. and N.H.; All authors have approved to publish the paper.

Funding

Not applicable.

Availability of data and materials

All data that support the findings of this study are available within the article.

Declarations

Competing interests

The authors declare no conflicts of interest.

Footnotes

Publisher’s note

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Contributor Information

Samy Selim, Email: sabdulsalam@ju.edu.sa.

Mohamed A. Amin, Email: mamin7780@azhar.edu.eg

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