Facile synthesis and optical characterization of selenium nanoparticles synthesized using Clitoria ternatea and Zingiber officinale: in vitro biomedical evaluation of antioxidant potential and antibacterial activity against caries-causing microbes

ABSTRACT

In this study, selenium nanoparticles will be produced with Clitoria ternatea and Zingiber officinale, and their efficacy as a natural anti-cariogenic agent will be assessed. To do this, a variety of analysis and evaluation techniques have been used, such as UV-Vis spectroscopy to characterize the synthesized selenium nanoparticles that has absorption band at 440 nm. XRD of the Se nanoparticle were analysed and were found to be crystalline in nature with particles size of 36 nm. SEM images confirmed that Se nanoparticles were flaky structured. The antibacterial activity test to assess the selenium nanoparticles activity against dental caries causing bacteria and the zone of inhibition of the nanoparticles were found. Cytoplasmic leakage assay was done to govern the range of damage instigated by the synthesized selenium nanoparticles to bacterial cell membranes and time kill curve assay represents the time taken by the synthesized selenium nanoparticles to kill the cariogenic bacteria.

KEYWORDS:

• Selenium nanoparticle
• clitoria ternatea
• zingiber officinale
• cytoplasmic leakage
• biomedical applications

Introduction

Nanoparticles are tiny particles with sizes in the range of 1–100 nanometres. They can be made from a wide variety of constituents, like metals, semiconductors, polymers, and ceramics [1]. Nanoparticles can be synthesized using various approaches, they can also be engineered with specific properties by controlling their size, shape, and surface chemistry. The properties of nanoparticles make them attractive for a wide choice of applications in areas namely medicine, electronics, energy, and environmental remediation [2]. In medicine, nanoparticles have been extensively studied for their potential use in drug delivery. Drug effectiveness and adverse effects can be improved by tailoring the exterior of nanoparticles to target particular cells or tissues. In addition, nanoparticles can improve the stability of drugs, allowing for controlled release over time [3,4].

Due to its higher bioactivity, interactions with proteins, excellent absorption capability, and minimized toxicity, along with its interdisciplinary roles in healthcare, therapeutic sciences, nanotechnology bioinformatics, and nanobiotechnology, selenium has attracted the most interest among the various types of nanoparticles [5]. Selenium has a critical role in moderating oxidative stress, minimizing the damage in cardiovascular illnesses, and diseases such as diabetes, cancer, and hypercholesterolaemia. Selenium is found in a variety of antioxidant enzymes and functioning protein molecules. On the other hand, selenium intake is essential for a variety of metabolic activities, and excessive intake causes selenium toxicity [6–9]. Selenium is also a vital micronutrient for plants, although it is only required in very small amounts, it is a trace element that plays a vibrant role in the growth and development of plants, as well as in their response to stress. Some of the important functions of selenium in plants include antioxidant defence, protein synthesis, abiotic stress tolerance, disease resistance, and nutrient uptake [10].

Several researchers have looked into the potential of selenium nanoparticles (SeNPs) to improve the therapeutic characteristics of plants. Clitoria ternatea (Butterfly pea) and Zingiber officinale (Ginger) are two plant species whose therapeutic properties have been extensively researched. Clitoria ternatea and Zingiber officinale are identified for their medicinal properties, like antioxidant, anti-inflammatory, antimicrobial activities, Memory and cognitive function, anxiolytic and antidepressant effects, wound healing, anti-diabetic properties, anticancer properties, cardiovascular health, liver protection, anti-ageing effects, anti-allergic properties, pain relief and immune boosters. These plants have been traditionally employed in Ayurvedic and Chinese medicine for various medicinal purposes. They have a wide range of potential medicinal uses. While more research is needed to fully understand its properties and potential benefits, it requires much prior experience in conventional medicine and shows promising results in modern studies [11–15]. Hence, this study formulated the potential of SeNPs in enhancing the medicinal properties of Clitoria ternatea and Zingiber officinale which can be contributed to the growing body of knowledge on the usage of nanotechnology in agriculture and medicine [16–19, p.26]. Hence, here we have reported the use of synthesized selenium nanoparticles from Clitoria ternatea and Zingiber officinale as an anti-cariogenic agent and assessed their toxicity to human cells.

Materials and methods

Preparation of plant extract

The well-dried Clitoria ternatea (Blue Tea) flowers and Dry Zingiber officinale (Dry Ginger) powder were collected. Clitoria ternatea dried flowers 2.5 grams were taken and Zingiber officinale powder 2.5 grams was taken and an extra 50 ml distilled water was added to each flask. For 10 minutes, the mixture was heated in a mantle at 50–60°C. After that, the filter paper from Whatman No. 1 was used to filter the heated mixture [20]. For the creation of selenium nanoparticles, filtered Blue Tea + Dry Ginger extract was kept in the refrigerator (Figure 1).

Figure 1. Extract of blue tea (Clitoria Ternatea).

Preparation of selenium nanoparticle solution

In 60 ml of distilled water, 20 mM of sodium selenite was synthesized. The extract was added to that 40 ml of filtered Blue Tea + Dry Ginger. This mixture spent 15 minutes in a digital ultrasonic cleaner and 30 minutes in a shaker to achieve uniform dispersion, which is a need in the synthesis of nanoparticles. A double-beam UV-visible spectrophotometer was used to constantly record the colour changes in the reaction mixture at various wavelengths between 250 and 650 nm. Centrifugation at 8000 rpm for 10 minutes separated the created Blue Tea + Dry Ginger extract-mediated selenium nanoparticles. The pellet of selenium nanoparticles that was produced was gathered and used for various characterization experiments [21].

Characterization of Se nanoparticles (blue tea + dry ginger)

The Morphology and spectral characterization studies were done to analyse and confirm the particle size, shape and nature of nanoparticles [22].

UV-Visible spectroscopy analysis

In the initial physicochemical characterization of the biosynthesized SeNPs, the reduction of selenium ions was examined using 3000 + from LABINDIA UV-Vis spectroscopy. It was used to characterize the synthesized nanoparticle solution between 250 and 650 nm. For graphical analysis, the findings were recorded at several time intervals, including 1 hour, 3 hours, and 24 hours.

X-ray diffraction (XRD) and FTIR analysis

The crystalline properties of the biosynthesized Se NPs were investigated using an X-ray diffractometer (Shimadzu Corporation (Japan); XRD-7000 X-ray diffractometer). On a device running at a voltage of 45 kV and an electrical current of 40 mA, the Cu K radiation source was employed with a wavelength of 1.5406 and a scanning angle of 2 in the scattering range of 1090°.

Fourier Transform Infrared (FTIR) spectroscopy examination in the 4000–400 cm⁻¹ range was performed using a Bruker FTIR spectrophotometer to investigate the role of different biomolecules that serve as capping, reducing and stabilizing agents in the synthesis of SeO₂ NPs.

Scanning electron microscopy

The air-dried Se NPs were placed on the stub using carbon adhesive that was fixed before being gold-sputtered coated. Using a scanning electron microscope (SEM) EVO18 (CARL ZEISS), the produced sample was examined to identify Se NP’s morphological events and elemental analysis properties.

Antibacterial activity

Selenium nanoparticles (blue tea + dried ginger) were investigated for their anti-cariogenic action against oral pathogens like Streptococcus mutans, Lactobacillus, Enterococcus faecalis, and Candida albicans. The zone of inhibition was identified for this activity using Mueller Hinton agar (MHA). MHA was made and sterilized at 120 lbs for 45 minutes. The sterilized plates were filled with media, which was then allowed to stabilize before solidifying. The test microorganisms were swabbed after the wells were cut with the appropriate cutter. Different concentrations of selenium nanoparticles were added, and the plates were then incubated for 24 hours at 37°C. The zone of inhibition was assessed following the incubation period [23].

Minimum inhibitory concentration

For performing Maximum bactericidal concentration, the different dilutions with pathogens in the MIC assay were swabbed on the surface of sterile Mueller Hinton Agar plates. Then, the plates were kept at 37°C for 24 hours. After incubation, the plates were counted and screened for the sum of colonies formed for each dilution [24].

Time kill curve assay

MHA broth was prepared, and sterilized and 6 mL was added to all the five test tubes. Bacterial suspension (Streptococcus mutans, Lactobacillus, Enterococcus faecalis and MRSA) was added to all five test tubes in the range of 5 × 10⁵ CFU/ml. The first three tubes contain the blue tea + dry ginger extract interceded selenium nanoparticles with three different concentrations and the fourth tube is considered as the growth control and the fifth tube as standard (Amoxyrite). The incubation was done under suitable conditions for varied time intervals (1 hr, 2hrs, 3hrs, 4hrs, 5hrs). Then the percentage of dead cells is calculated at a wavelength of 540 nm at regular time intervals [25].

Cytoplasm leakage assay

Protein, DNA, and K⁺ ion leakage in the cytoplasm were all examined. 50 ml of sterile NB were mixed with 0.1 OD cultures of Streptococcus mutans, Lactobacillus, Enterococcus faecalis, and Candida albicans, and the mixture was then incubated at 37°C for 24 hours. The broth was centrifuged for 10 minutes at 5000 rpm. The supernatant was removed, the cell pellet was collected, and cells were then resuspended in 50 ml of saline solution with 100 g ml⁻¹ of TiO₂ NPs (of both 8–10 nm and 90–100 nm size) as the test sample, while cells without NPs in 50 ml of saline were kept as the control, and both were incubated overnight at 37°C. It was centrifuged for 10 minutes at 5000 rpm. The supernatant was gathered for the Bradford protein test, DNA quantification using the diphenylamine (DPA) method, and K⁺ content measurement using flame photometry [26].

Antioxidant assay

DPPH Assay (2, 2-diphenyl-1-picryl-hydrazyl-hydrate)

The antioxidant capacity of biogenic synthesized selenium nanoparticles was evaluated using the DPPH assay. Diverse quantities (10–50 g/ml) of Blue Tea + Dry Ginger extract blocked the action of selenium nanoparticles. They were combined with 450 l of 50 mM Tris-HCl buffer (pH 7.4) and 1 ml of 0.1 mM DPPH in methanol, and incubated for 30 minutes. Later, based on the absorption at 532 nm, the decrease in the number of free radicals caused by DPPH was measured. Butylated hydroxytoluene was used as the control [27]. The following equation was used to calculate the percentage of inhibition:

% inhibition is calculated as (control absorbance – test sample absorbance)/Absorbance of control * 100.

H₂O₂ (hydrogen peroxide)

Minor modifications were made to the Halliwell method [28] to carry out the assay. Every solution was made from scratch. The reaction mixture, which had a volume of 1.0 mL, contained 100 mL of 28 mM 2-deoxy-2-ribose (dissolved in phosphate buffer, pH 7.4), 500 µL of a solution of different concentrations of blue tea + dry ginger extract that served as an intermediary for selenium nanoparticles (10 to 50 µL), 200 µL of 200 µM Fecl3 and 1.04 mM EDTA (1:1 v/v),100 µL H₂O₂ (1.0 mM) and 100 µL ascorbic acid (1.0 mM). The TBA reaction was used to gauge the degree of deoxyribose breakdown following an hour-long incubation time at 37°C. Using a blank solution as a reference, calculate the absorbance at roughly 532 nm. A positive control was utilized, which was vitamin E.

Cytotoxic effect

2 grams of iodine-free salt were measured and dissolved in 200 ml of clean water. Six well ELISA plates were loaded with 10–12 ml of saline water. 10 nauplii (5, 10, 20, 40, 80, and control) were added to each well. After that, the nanoparticles were added at the desired dosage, and the plates were incubated for 24 hours [29]. Following incubation, the existence of live nauplii was counted on the plates, and the number was calculated using the formula below:

Total number of dead nauplii = total number of dead nauplii + total number of livings nauplii 100.

Results and discussion

Preparation of plant extract of Clitoria ternatea (blue tea) - Zingiber officinale (ginger) - synthesis of selenium nanoparticles

The first step in the shaping of nanoparticles was visual colouration. After the incubation period, it was noticed that the colour turned brown. Figures 2 and 3 depict the 48-hour time-dependent colour shift that occurred when Clitoria ternatea (blue tea) and Zingiber officinale (ginger) were combined in a reaction with sodium selenite. The mixture was initially yellow during the primary reaction step, but with time it turned brown. There was no further change in hue after 48 hours of incubation. This brown colour may be attributable to the selenium nanoparticles’ stimulation of the surface plasmon vibrations, providing useful spectroscopic proof of their production.

Figure 2. Extract of dry ginger (Zingiber Officinale).

Figure 3. Preparation of SeNPs using plant extract Clitoria ternatea (blue tea) + Zingiber officinale (ginger).

Characterization of selenium nanoparticles

The synthesis of SeNPs is supported by the prominent peak in the UV-visible spectrum at 440 nm (Figure 4). Over time, the peak intensity grew stronger. After the reaction had been going for 12 hours, no significant increase in peak intensity was seen. Due to SeO₃^2- to Se⁰ has been reduced over time, the peak amplitude has grown, indicating that SeO₃^2- to Se⁰ conversion has reached its maximum. One of the most used techniques for determining SeNPs’ structural characteristics is UV-vis spectroscopy. UV-vis spectroscopy may typically be used to determine the size and form of the monitored aqueous suspension NPs. Our findings show that extracts of Clitoria ternatea (blue tea) and Zingiber officinale (ginger) produce SeNPs. Unbound electrons in metal nanoparticles form a surface plasmon resonance (SPR) absorption band as a result of the mutual oscillation of metal nanoparticle electrons in resonance with light waves, and this band was used for further characterization and biological applications. A 440 nm peak in the measurements, according to data and spectrophotometers, indicates Se NP performance [30]. Previous research has found that the strong plasmon peak of spherical Se-NPs may be seen in the UV-Visible spectra between 250 and 400 nm [31].

Figure 4. UV-vis spectroscopic analysis of Clitoria ternatea (Blue Tea) and Zingiber officinale (ginger) mediated selenium nanoparticles at different time intervals and wavelength from 250 nm to 650 nm.

X-Ray diffraction of selenium nanoparticles

The pure hexagonal phase in Se crystals of lattice parameters a = 4.366A0 and c = 4.9536A0 were reflected in the diffraction peaks at 36.7, 42.8, 63.2, and 76.4, respectively (100), (101), (110), and (102) (JCPDS Card No. 06–362). Figure 5 depicted a selenium XRD picture. Utilizing powder X-ray diffraction spectroscopy, the crystal structure of the selenium nanoparticle was examined. The Se NP diffraction peaks with values of 32.84, 38.18, 44.34, 54.92, 64.46, and 77.38°Correspond to the planes’ in the Miller nomenclature: (110) (111), (200), (220), and (311). These resemble JCPDS card No. 04–0783, which describes the cubic structural structure of Se NPs, in many ways. The peaks gained were due to the influence of active phytochemical contents in the extract [32]. The XRD diffraction investigation revealed that selenium ions are present and that Se NPs are crystalline. The Debye-Scherer formula (D = (k/cos)), where D stands for the crystalline size (nm), denotes the X-ray wavelength (0.1541 nm), denotes the angular line full width at half-maximum (FWHM) of the peak (in radians), and denotes the Braggs angle (in degrees), was used to calculate the average crystal size of the nanoparticles. For the green synthesis of Se NP, the crystal size of the produced Se NPs was determined to be 36.64 nm. Uncalcined biosynthesized nanoparticles retain the character of the parent components (organic compounds), and the maximum carbon concentration results in an amorphous form of the material. In this scenario, the nanoparticle derived from a high-volume percentage of plant extract has a more amorphous structure than the nanoparticle derived from a low proportion of plant extract. Otherwise, the size of the nanoparticle synthesized with a high proportion of plant extract is lower than the size of the nanoparticle synthesized with a low percentage of plant extract. This is because a higher concentration of plant extract has a higher concentration of organic capping (reducing) ingredients. This means that the extract has more organic components [33].

Figure 5. XRD of Se NPs from C. Ternata+ Z. officinale.

Scanning electron microscopy of Se nanoparticles

The form of the selenium nanoparticles produced through the biological reduction process was examined using a scanning electron microscope. The nanoflakes of dispersed selenium nanoparticles were seen in SEM pictures. The growth of selenium nanoparticles was confirmed by the SEM image of produced selenium nanoparticles, which was visible over the tiny implants. Figure 6 displays a SEM view of selenium nanoparticles. These tiny nanoparticles will be favourable to inhibit rod and spherical shape microorganisms [34,35].

Figure 6. SEM of Se NPs from C. ternata+ Z. officinale.

FTIR spectroscopy

FT-IR spectroscopy was used to characterize the synthesized SeO₂ nanoparticles to identify the functional groups that were present in the nanoparticles that were synthesized (Figure 7). According to our findings, there are more than four functional groups present in the nanoparticles that we synthesized. The presence of C=C stretching bonds, strong C=O stretching bonds, and strong C-H stretching bonds in nanoparticles was indicated by peak values of 1637.43, 1725.60, and 3337.49 correspondingly. The bending and rocking vibrational modes of C – H and C – C bands are responsible for the appearance of C – H bonds in gelatin below 1500 cm⁻¹, which appeared in the range of 2800–3000 cm⁻¹. According to these findings, gelatin and glucose species managed to attach themselves to the surface of the nanoparticles that were synthesized. As a result, the presence of bioactive phytochemicals in the extract, such as polyphenols, flavonoids, tannins, amino acids, and sugars, may be involved in the bioreduction of Se NPs forms corona, as confirmed by FTIR analysis, where it also acts as a surface stabilizing agent for Se NPs while also improving biocompatibility. The capping avoids particle agglomeration and so stabilizes them in the medium [36–38].

Figure 7. FTIR of Se NPs from C. Ternata+ Z. officinale.

Antibacterial activity of Se NPs

The antibacterial action of selenium nanoparticles mediated by Clitoria ternatea (Blue Tea) and Zingiber officinale (Ginger) in a zone of inhibition against C. albicans, L. acidophilus, E. faecalis, and S. mutans is demonstrated as shown in Figure 8. S. mutans had the maximum zone of inhibition, which was immediately followed by C. albicans, L. acidophilus, and E. faecalis. When compared to the antibiotic controls, selenium nanoparticles showed strong antibacterial efficacy against bacteria that cause tooth decay at all doses. This is most likely due to Gram-negative bacterial cell membranes having a larger negative charge than Gram-positive bacteria. As a result, nanoparticles are more likely to cling to Gram-positive bacteria surfaces and kill them [32].

Figure 8. Antibacterial activity of Se NPs against caries-causing microbes (C. albicans, L. acidophilus, E. faecalis and S. mutans) at different concentrations and zone of inhibition was measured in millimetres.

Minimum inhibitory concentration

By using the micro-well dilution technique, the MIC -minimum inhibitory concentration of selenium nanoparticles against pathogenic bacteria was calculated as represented in Figure 9. The positive control tubes with selenium nanoparticles showed turbidity after 24 hours of aerobic incubation at 37°C, indicating the development of bacteria. While there was no turbidity at concentrations of 25, 50, and 100 µg/ml, showing that bacterial growth was being inhibited. It was due to the thinner peptidoglycan layer and the presence of porins, Se-NPs exhibited a higher inhibitory effect [31]. The antibacterial efficacy of selenium against Staphylococcus is supported by prior research, which also suggests that selenium can stop bacteria from forming biofilms and that SeNPs have lethal effects [39, 40]. According to the study, as synthesized SeNPs could be a highly effective antibacterial agent in the biomedical area.

Figure 9. Minimum inhibitory concentration of Se NPs.

Time kill curve assay

The time-dependent bactericidal activity of Clitoria ternatea (Blue Tea) and Zingiber officinale (Ginger) mediated selenium nanoparticles is shown in Figure 10(a–d) this assay was done with selected strain suspension (L. acidophilus, Streptococcus mutans and Enterococcus faecalis). The time-kill experiment provides details on the processes and behaviours of the examined antibiotics when they come into contact with the bacteria. Additionally, it is possible to determine if the substances are bacteriostatic or bactericidal. When the bacterial concentration falls by 3 log10 of the initial inoculum concentration, a substance is deemed bactericidal. Bacterial death can also be measured in terms of time and antibiotic concentration [40][]. The actual reduction in cell viability (CFU/mL) at intervals of 1, 2, 3, 4, and 5 hours for every isolate was used to assess the bactericidal activity. At five hours, the growth stopped. On L. acidophilus, Streptococcus mutans, and Enterococcus faecalis, the antibacterial property of SeNPs stabilized with Clitoria ternatea (Blue Tea) and Zingiber officinale caused bacterial harm to the peptidoglycan layer of the bacterial cell membrane, where the cytosol content leaks out and causes cell death [41].

Figure 10. Time kill curve assay of SeNPs (Clitoria ternatea + Zingiber officinale) against (b) E. faecalis (c) L. acidophilus (d) S. mutans.

Cytoplasm leakage assay

According to past studies, selenium nanoparticles cause pathogenic bacteria’s membrane integrity to be disturbed. We measured the quantity of cytoplasmic material (DNA and protein) that the microorganisms released in order to obtain a deeper understanding of the impacts of this chemical [42,43]. Spectroscopic analysis was used to compare the amounts of external DNA and protein in the supernatants of bacteria treated with NP (MIC₅₀) and left untreated in PBS buffer. The findings of this study showed that all of the bacteria studied lost intracellular content when their resting cells were exposed to NPs. When compared to a control group of all bacteria, the analysis of released protein and DNA components in the filtrate from bacterial cells was extensive at various concentrations (10, 20, 30, 40, and 50 µg/ml). The outcomes are displayed in Figure 11. The DNA produced by the bacteria at various concentrations varied between them in statistically significant ways. S. mutans had the maximum amount released, followed by C. albicans, L. acidophilus, and E. faecalis. The DNA molecules are condensed and lose their capacity to proliferate as a result of the contraction of the cytoplasm membrane from the cell wall. It is also advised that bacterial proteins be deactivated. Finally, when the membrane shrinks, the likelihood of nutrients entering the cell for continued growth and proliferation decreases. In other words, the selenium particles in the hybrid structure would interact with the bacteria constantly and homogeneously, assuring long-term antibacterial effects [44].

Figure 11. Cytoplasm leakage assay of SeNPs (Clitoria ternatea + Zingiber officinale).

Antioxidant activity

DPPH assay

Figures 12 depict DPPH’s antioxidant action. Selenium nanoparticles extract from Clitoria ternatea (Blue Tea) and Zingiber officinale (Ginger) scavenges DPPH radical in a dose-dependent manner (10 µL to 50 µL). Despite an increase, the percentage reduction in selenium nanoparticles is still much lower than that of conventional ascorbic acid. Consequently, concerning the antioxidant property of the reference standard, selenium nanoparticles are highly effective [44].

Figure 12. DPPH assay of SeNPs (Clitoria ternatea + Zingiber officinale).

H₂O₂ (hydrogen peroxide) assay

Figures 13 depict the hydroxyl radical scavenging activity of selenium nanoparticle extracts mediated by Clitoria ternatea (blue tea) and Zingiber officinale (ginger). Selenium nanoparticles’ capacity to scavenge hydroxyl radicals in this investigation exhibits action in a dose-dependent way. The outcomes are contrasted with ascorbic acid which is typically used. Lipids, proteins, and DNA are just a few of the nearby biomolecules that the hydroxyl radical severely damages. In these circumstances, green synthesized selenium nanoparticles may be used because they are effective hydroxyl radical scavengers. The primary chemicals responsible for antioxidant action are all phenolic compounds [44].

Figure 13. Hydroxyl radical scavenging activity of Clitoria ternatea (Blue Tea) and Zingiber officinale (ginger) mediated selenium nanoparticles extract.

Determination of cytotoxic effect

Selenium nanoparticles’ cytotoxic activity demonstrates that all of the introduced shrimps remained alive in the control. On day 1, every shrimp was alive. On day 2, 90% of the shrimp in the 20 µg/ml and 80% of the shrimp in the 40 µg/ml nanoparticle wells were still alive. Only 70% of the shrimp in the well containing an 80 µg/ml nanoparticle concentration were still alive beyond 48 hours. It’s displayed as a bar graph (Figure 14). Therefore, it is clear that selenium nanoparticles have negligible cytotoxicity against shrimp and non-toxic nanomaterial [45].

Figure 14. Cytotoxic effect of SeNPs (Clitoria ternatea + Zingiber officinale).

In contrast to metal nanoparticles such as silver and copper, elemental selenium is typically regarded as insoluble in aqueous settings. There is a belief that SeNPs have the potential to undergo conversion into organic forms, such as seleno amino acids and selenoproteins, as a result of interactions with microorganisms [46,47]. Selenium has the potential to displace sulphur in sulphur-containing ammonic acids, such as cysteine and methionine, due to their chemical similarities [48]. An overabundance of selenoproteins has been found to result in the production of reactive oxygen species (ROS), which can induce DNA damage, alter protein structure, and impair enzyme activity in comparison to silver and copper nanoparticles [49].

Conclusion

The synthesis and characterization of selenium nanoparticles utilizing Clitoria ternatea and Zingiber officinale showed promising results in terms of their potential as a natural anti-cariogenic agent. The synthesized selenium nanoparticles were found to possess antioxidant and antibacterial properties and were operative in inhibiting the development of cariogenic bacteria. The various characterization techniques used in this project, such as UV-VIS, XRD and SEM spectroscopy provided important information on the properties of the synthesized selenium nanoparticles, including their size, shape, crystalline nature, and chemical composition. The evaluation of minimum inhibitory concentration, time-kill curve, cytoplasmic leakage, protein leakage, and cytotoxicity provided insight into the efficacy and safety of the synthesized selenium nanoparticles. The use of natural sources for the synthesis of selenium nanoparticles provides a sustainable and environmentally friendly approach to the production of nanomaterials. The synthesized selenium nanoparticles exhibited a broad-spectrum antibacterial activity, which suggests that they may be effective against other bacterial infections as well. The antioxidant activity of the synthesized selenium nanoparticles could potentially make them useful in preventing oxidative stress-related diseases. They also showed low cytotoxicity, which is a promising finding for their potential use as a therapeutic agent. The results of this project demonstrate the importance of interdisciplinary research, as it involved the fields of chemistry, biology, and materials science. This article highlights the need for further research into the synthesis and characterization of selenium nanoparticles, as well as their potential applications in various fields. The findings of this project suggest that the produced selenium nanoparticles using Clitoria ternatea and Zingiber officinale have potential for use in the progress of natural and safe antimicrobial and antioxidant agents.

Acknowledgments

Authors are thankful to the Researchers Supporting Project number (RSPD2024R728), King Saud University, Riyadh, Saudi Arabia.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.

Disclosure statement

No potential conflict of interest was reported by the author(s).