ZnO Nanoparticles Synthesized using Aerial Extract of <i>Ranunculus multifidus</i> Plant: Antibacterial and Antioxidant Activity

Yilma, Terfo; Kassaw, Mikyas; Murthy, H. C. Ananda; Dekebo, Aman

doi:https://doi.org/10.1155/2023/8825762

Journal of Nanomaterials

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Abstract Introduction Materials and Methods Results and Discussion Conclusion Data Availability Conflicts of Interest Acknowledgments References Copyright Related Articles

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Sustainable Synthesis of Nanomaterials for Environmental Remediation and Biomedical Applications: A Way Towards Circular Economy

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Volume 2023 | Article ID 8825762 | https://doi.org/10.1155/2023/8825762

ZnO Nanoparticles Synthesized using Aerial Extract of Ranunculus multifidus Plant: Antibacterial and Antioxidant Activity

Terfo Yilma,¹Mikyas Kassaw,¹H. C. Ananda Murthy,^1,2and Aman Dekebo^1,3

Academic Editor: Jagpreet Singh

Received28 Dec 2022

Revised08 Feb 2023

Accepted25 Apr 2023

Published21 May 2023

Abstract

The present work reports the synthesis of zinc oxide nanoparticles (ZnO NPs) by applying an aqueous aerial extract of Ranunculus multifidus plant. The thermogravimetric analysis revealed that the prepared ZnO NPs are stable from 480 to 800°C. The diffraction study confirmed the hexagonal wurtzite structure for the synthesized ZnO NPs with the typical crystallite sizes of 47.92, 22.70, and 15.35 nm the volume ratios (extract to precursor) of 1 : 1, 3 : 2, and 2 : 3, respectively. The experimentally deduced E_g values are 1.82, 3.1, and 2.57 eV for 1 : 1, 3 : 2, and 2 : 3 ZnO NPs, respectively. The spherical and rod-like morphologies were confirmed for the NPs by the images taken using electron microscopy. The reducing agents in the aqueous extracts of R. multifidus converted the ionic zinc to zinc nanoparticles, and these NPs exhibit credible antibacterial effects against tested bacterial species. The biosynthesized ZnO NPs revealed significant antibacterial activity against Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. The order of the antibacterial potential of the NPs was found to follow the order: S. aureus (17.10 ± 0.45 mm) > B. subtilis (16.10 ± 0.15 mm) > E. coli (14.5 ± 0.32 mm) > P. aeruginosa (13 ± 0.0 mm). The antioxidant activities of the produced ZnO NPs in various ratios showed the potentiality of phytochemicals to scavenge the free radicals, which is encouraging for the discovery of novel compounds for the treatment of cancer diseases.

1. Introduction

The novel method of plant-mediated synthesis of nanoparticles (NPs) has numerous applications in medicine, pharmacy, industry, and agriculture. Due to their toxicity, conventionally produced NPs are only occasionally used in the therapeutic setting. We are currently observing the growth and development of a new interdisciplinary scientific discipline called nanoscience [1–8]. There are many methods for the production of nanoparticles, but green synthesis is one of the most popular since it is safe for the environment, nontoxic, cheap, and extremely pure. A chemical wet method was also used to prepare nanomaterial, especially with ZnO–Ag and ZnO–Au as nanocomposite materials by using the precipitate method [9]. Noble metal and metal oxide nanoparticles including silver, copper, gold, platinum, CuO, and Ag₂O were produced using many plant extracts by adopting the green methodology. But among many metal oxide NPs, zinc oxide nanoparticles (ZnO NPs) shine in the biological and pharmaceutical industries.

In recent years, different industrial sectors, such as pharmaceuticals, cosmetic, and concrete industries, have begun to use zinc oxide as some key ceramic materials instead of the microbiological, textile, and automotive industries. Due to the worrisome rise in the incidence of bacterial infectious illnesses and their resistance to most first-line antibiotic agents, antibacterial therapy has proven challenging [10]. In the twenty-first century, this poses a serious risk to human health, necessitating urgently continuing research to find agents with stronger antibacterial and broad-spectrum activities.

Therefore, finding novel drugs to treat disease and inflammations without causing significant negative impacts on patients was the current project on which all researchers were engaged. In nanoscale formulations, ZnO NPs are now being studied as potential allies of antioxidant and antibacterial medicines. The ZnO NPs have anticancer properties and are frequently used to treat a variety of skin problems [6]. Furthermore, ZnO NPs have been used as a recent preferable tool in drug delivery and sensing horizons. Traditional medicines in a different country have a long history and are mostly based on rich, though unstandardized, pharmacopeia made primarily from plants that are used by patients at home for self-administration as well as traditional healers [11]. Due to its ethnomedicinal benefits that have been reported in various locations by different herbalists and researchers, Ranunculus multifidus, one of the traditional medicinal plants primarily found in Africa, particularly Ethiopia, is taken into consideration in this work [12]. The bio-diversified species of flowering plant R. multifidus, often known as buttercup in South Africa, belongs to the Ranunculaceae family. With the exception of West Africa, Madagascar, and the Arabian Peninsula; it is indigenous to Sub-Saharan Africa [12]. Anti-rheumatism, intermittent fever, and rubefacient are the three traditional uses of Ranunculus species [13]. In a similar situation, R. multifidus was also traditionally used to cure several diseases. The most widely adopted traditional uses includes treatment of TB [14], infertility, blood cleansing [15], eye infection [16], shingles and sores, malaria [17], and others. This traditional use of the plant might emanate from its phytochemical components such as flavonoids, saponins, alkaloids, glycosides, terpenoids, anthocyanins, and quinones. Despite the plants having a wide range of conventional applications, there are no reports of this amazing herb being used to synthesize nanoscale materials. The main objective of the current study was to explore or search the application of R. multifidus plant aerial extract as a capping and reducing agent for the synthesis of ZnO NPs because it is a rich source of bioactive compounds and investigate their antibacterial and antioxidant activity in light of the significance of green nanoparticles and research gap toward the plant mentioned. The plant extract transforms ionic zinc to metal zinc oxide nanoparticles by acting as a reducing and capping agent. Therefore, the antibacterial activity of synthesized ZnO NPs was evaluated against Gram-negative and Gram-positive bacterial strains namely, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis. In addition, the radical scavenging activities of synthesized NPs were evaluated using the DPPH methods. The result of the present work revealed that the synthesized nanoparticles exhibited significant antibacterial activities, especially on Gram-positive pathogenic strains. The radical scavenging activities of ZnO NPs are promising for the disease-associated ROS. To the best of our knowledge, this research was the first and novel work using this plant and gives a great clue for further works, especially using nanohybrid materials to enhance the potency of nanoparticles for different biomedical applications.

2. Materials and Methods

2.1. Plant Material Collection and Identification

The aerial parts of the plant R. multifidus were picked from Bekoji, Oromia region, Arsi zone. The authentication of the plant was done at the National Herbarium of Ethiopia, Addis Ababa University (voucher no. TY-001).

2.2. Preparation of Plant Extracts (Broth Solution)

R. multifidus fresh plant material was collected, completely cleaned with distilled and tap water to get rid of the dust, and then dried in the shade to get rid of any remaining moisture. The powdered plant materials were sieved using 180 m size sieves after being pounded into powder. The sample’s aqueous extracts were made by boiling 50 g of finely ground powder for 60 min at 80°C with constant stirring. After filtering the extract using Whatman no. 1 filter paper and allowing it to cool to room temperature, the extract was kept in a refrigerator at 4°C for future experimental usage [5].

2.3. Synthesis of ZnO NPs

Three different ratios R. multifidus plant extract and precursor salts were used: 1 : 1, 3 : 2, and 2 : 3, 50 mL, 60 mL, 40 mL of extract and 50 mL, 40 mL, and 60 mL of 0.2 M zinc acetate (Zn(O₂CCH₃)₂(H₂O)₂), were mixed with 50 mL of 0.2 M NaOH, respectively.

A yellow precipitate formed after the three solutions were continuously for 2 hr at 800 rpm using a magnetic stirrer [18]. The precipitate was periodically cleaned in ethanol and distilled water to get rid of any impurities left behind, and they were then oven dried at 100°C for an hour. Using a blender, the produced dried light yellow color powder was prepared for further characterization and applications. The overall synthesis process was depicted in Figure 1.

2.4. Characterization of Biosynthesized ZnO NPs

A simultaneous DTA-TGA (DTG-60H, Shimadzu Co., Japan) study was used to examine the thermal stability and decomposition followed by the weight loss of the prepared ZnO NPs measured at the heating rate of 10°C/min. An ultraviolet–visible (UV–vis) spectrophotometer was utilized to identify the surface plasmon resonance (SPR) peak. Using Fourier transform infrared spectroscopy, the potential phytochemicals causing the reduction of metal salts into ZnO NPs were examined (FTIR). The external morphology, surface features, and elemental compositions of ZnO NPs were analyzed using -microscopic methods combined with energy-dispersive X-ray analysis (SEM-EDX).

2.5. Antibacterial Investigation of Synthesized ZnO NPs

The antibacterial activity of biosynthesized ZnO NPs was performed using the agar disc diffusion method against human infectious, B. subtilis, E. coli, P. aeruginosa, and S. aureus, bacterial strains which were American Type Culture Collection (ATCC). In nutrient broth, cultures of the test organism were grown to the late logarithmic phase. Aliquots of 100 μL from the cultures were spread on solidified nutrient agar plates. The plates were coated with the 100 μL solution of 50, 75, and 100 μg/mL concentration of ZnO NPs solution impregnated disc (6 mm). The plates were then placed upside down in the incubator at 37°C for 24 hr. To ensure that bacterial cells and agar were mixed equally, the plates were gently shaken. After the incubation period got over, different levels of zonation formed around the disc were measured using a caliper. This procedure was done for all ratios of biosynthesized ZnO NPs against standard bacterial strains.

2.6. DPPH Radical Scavenging Assay

DPPH radical scavenging assay was conducted according to the previously described standards with little modification, the DPPH test was used to evaluate the free radical scavenging activity (RSA) of the plant extract and synthesized ZnO NPs of various ratios (1 : 1, 2 : 3, and 3 : 2) salt to plant aqueous aerial extract [19]. The 1,1-diphenyl-2-picryl hydrazyl (DPPH), a persistent free radical, can decolorize in the presence of antioxidants, which is the basis for the DPPH antioxidant assay. The odd electron in the DPPH radicals is what causes the absorbance at 517 nm as well as the apparent to deep purple color. A decrease in the absorbance of the reaction mixture indicates significant free radical scavenging activity of the extract or synthesized NPs. The data were compared with those obtained with the reference ascorbic acid. In the process, the synthesized ZnO NPs of different ratios (1 : 1, 2 : 3 and 3 : 2) salt to plant extract ration were dissolved in methanol to afford 1 mg/mL. It was serially diluted in methanol to yield a 500, 250, 125, and 62.5 μg/mL concentration. Four milliliters of DPPH (0.004% DPPH in MeOH) was added to 1 mL of each concentration to make 100, 50, 25, and 12 μg/mL. Then all the samples prepared were incubated in an oven at 37°C for 30 min and then absorbance was recorded at 517 nm using a UV–vis spectrophotometer. A reaction solution without DPPH was used as blank and DPPH solution as a control. Ascorbic acid was used as standard. The percentage inhibition was calculated using the Equation (1):

The IC₅₀, or the concentration of the test substance needed to cause a 50% reduction in absorbance from that of the control solution, is another way to express the sample’s ability to scavenge DPPH radicals [20]. The concentration vs. RSA% graph was produced using Excel, and the IC₅₀ values were calculated from it and interpreted as the smallest IC₅₀ values mean the strongest radical scavenging concentration of extracts.

3. Results and Discussion

3.1. Synthesis ZNO NPs

Using R. multifidus and zinc acetate dihydrate salt as a precursor, ZnO NPs were synthesized. The aerial extract from R. multifidus was used as a reducing and capping agent. The presence of flavonoids, phenolic compounds, tannins, saponins, anthraquinone glycosides, reducing sugars, phytosterols, steroids, terpenoids, and glycosides in the plant extracts revealed the phytochemical screening of the extracts. The information on the phytochemicals in the extract is provided in Table 1. The three main processes that go into making NPs, metal ion reduction, cluster formation, and nanoparticle development, are thought to be extremely important. The tautomeric conversion of polyphenols from their enol form to their keto form can be expected to reduce the zinc ions to zinc NPs.

Additionally, the enzymes in the aerial extract of the plant assist the metal ions to create an enzyme–substrate complex, which results in the production of zinc with a protein cap [21], and it is known that the phenolic chemicals serve as ligands, bind to metal ions, decrease, and cap those ions to create nanoparticles. A prior study had also claimed that these ligands also function as regulators of particle size [21, 22]. The great propensity of phenolic compounds to chelate metals is the main cause of their antioxidant function. The hydroxyl and carboxylic groups found in phenolic compounds have a very high propensity to attract metal ions. In solution, the phenolic chemicals interact with metal ions, assisting in the nucleation and production of ZnO NPs.

3.2. Characterization of Synthesized ZnO NPs

The synthesized ZnO NPs were characterized by using the TGA and its derivative (DTA), UV–vis-DRS, FT-IR, XRD, and SEM techniques.

3.2.1. Thermogravimetric Analysis (TGA) and Its Derivative (DTA)

The sample’s mass loss is shown by the TGA curve, while the process’s energy gain or loss is shown by the DTA curve [5]. Figure 2 depicts the thermal analysis plots of the uncalcined ZnO NPs (1 : 1).

The three primary steps were observed from the ZnO NPs TGA curves; 6.89% of the water molecules in the first curve, in the temperature range from 20 to 250°C, reveal the dehydration of adsorbed water. In the second breakdown loss, which occurs as the temperature rises from 240 to 480°C, 10.75% of Zn (OH)₂ transforms into ZnO NPs. The remaining decomposition losses from 480 to 800°C are not displayed in the third stage. According to the evidence of breakdown losses, residual surfactants, or steady heat follow, ZnO NPs were found to be thermally stable from 480 to 800°C. The outcome of this investigation was consistent with earlier published research [23]. As a result, for all other manufactured ZnO NPs throughout this experiment, calcination temperatures of up to 480°C were used.

3.2.2. XRD Analysis

X-ray diffraction technique was utilized to explore the structure and crystallite size of the synthesized ZnO NPs. Figure 3 displays the diffraction pattern of ZnO NPs made from zinc acetate precursor and R. multifidus plant aerial extract in three different volume ratios, such as (1 : 1), (2 : 3), and (3 : 2). The lattice planes (100), (002), (101), (102), (110), (103), and (112) are successively represented by the 2θ values, 32.01°, 34.50°, 36.25°, 47.71°, 56.71°, and 68.00° in the XRD analysis of ZnO NPs. All the diffraction peaks recorded for the NPs were discovered to be in compliance with the literature report [5, 23]. The findings support the synthesis of pure ZnO NPs without the creation of any secondary phase and are in good accord with Miller index values and the hexagonal wurtzite structure of ZnO (JCPDS card no. 036-145). The evaluation using Scherer’s formula (Equation (2)) revealed the typical crystallite size of 47.92, 22.70, and 15.35 nm for ZnO NPs prepared in the volume ratios of (1 : 1), (3 : 2), and (2 : 3), respectively.where D is the crystallite size, K is 0.9 diffraction constant (shape factor), λ is 1.54 (X-ray source wavelength), β is the full width at half maximum intensity (FWHM) in radians, and θ is the angle of diffraction.

Table 2 displays the estimated and measured XRD parameters for crystallite size. The calculated and measured crystallite size XRD characteristics are shown in Table 2. The results revealed that the ratio of the plant extract grew from 1 : 1 (47.92 nm) to 3 : 2 (22.7 nm) and to 2 : 3 (15.35 nm) volume ratios, the average crystal size of ZnO NPs were found to increase. Since a greater amount of the plant extract was used during the synthesis process, it results in effective capping and stabilization of the synthesized nanoparticles and hinders the process of aggregation. This precisely matches the previously stated method of the production of ZnO NPs [24].

3.2.3. SEM Analysis

SEM was used to examine the surface morphological characteristics of the ZnO NPs. The micrographs are presented in Figure 4(a)–4(c). Figure 4(a) displays a micrograph of ZnO NPs produced using a 1 : 1 volume ratio of plant aerial extract and zinc acetate salt. ZnO NPs with a 1 : 1 volume ratio appeared as a honeycomb-like structure in the SEM picture. The increase of the nanoparticles’ size and the accumulation of zinc acetate and plant extract on their surface are what caused it to occur. Due to the increased concentration of zinc ions followed by aggregation on the surface for ZnO NPs in 3 : 2 volume ratios resulted in a road-like form and the ZnO NPs with 2 : 3 volume ratio, exhibited a spherical form [25]. According to the current work and earlier literature findings, the ability of the extract to serve as a cap during the nucleation, aggregation, and formation of NPs was what caused the reduction in size upon increasing plant extract concentration [25].

(a)

(b)

(c)

3.2.4. Fourier Transform Infrared (FTIR) Analysis

Figure 5 shows the FTIR spectra of ZnO NPs and aerial extract of R. multifidus plant. Biomolecules were found in the extract and NPs by FTIR spectral analysis. The major peaks that were detected at 3,421 and 2,943 cm⁻¹ represent, sp³ C–H stretching vibrations and the OH stretching vibration of carboxylic acid, respectively. The vibration of the CN moiety from the protein molecule of the extract is responsible for the peak at 2,151 cm⁻¹. The peak at 1,621 cm⁻¹ is related to carbonyl groups that have undergone C=O stretching and may be acidic carbonyl groups. Sharp peaks around 1,329 cm⁻¹ indicate the existence of the carboxylic acid COO– group. Additionally, it was demonstrated that the amine and carboxylate groups in the R. multifidus aerial extract were in charge of adhering to the surface of ZnO and stabilizing the biosynthesized ZnO NPs as well as fitting the prior literature study [21]. Around 1,044 cm⁻¹, the C–O–C stretching becomes visible. The peak at around 552 cm⁻¹ can be attributed to the stretching vibration of the Zn–O bond. The bending vibrations of Zn–O–H bonds resulted in a tiny peak at 683 cm⁻¹.

3.2.5. UV–Vis–DRS Spectral Analysis

Diffuse reflectance spectroscopy (DRS) was used to assess the optical energy band gap (E_g) values of the synthesized ZnO NPs in volume ratios (1 : 1, 2 : 3, and 3 : 2) as shown in Figure 6. The sample’s reflectance spectrum was found to be highly reflective between 447 and 456 nm in the UV spectrum, attaining a practically constant value thereafter, and the results were consistent with the report [26, 27]. The Kubelka–Munk formula (Equation (3)) was applied to convert the measured reflectance to absorbance.where A is the transition probability, hν is photon energy and the exponent factor, n is the nature of the optical transition (n can take up the values ½ or 2).

(a)

(b)

The bandgap energies (E_g) of synthesized ZnO NPs with the volume ratios of (1 : 1), (2 : 3), and (3 : 2) were 1.82, 3.1, and 2.57 eV, respectively, and were calculated based on Equation (3); the fraction of reflectance (F), the photon energy (hν), and direct bandgap energies (E_g). ZnO (2 : 3) NPs have a higher E_g value than the other two volume ratios. On the other hand, the 2 : 3 ratio’s average crystallite size from XRD shows that it is smaller than the other counterpart. As the concentration of the extract increases, the increased concentration of the phytochemicals stabilizes small particles by effectively capping them. This intern accounts for the decreased aggregation effect. If the particle size is very small, light interacts with the samples instead of being absorbed by it, with some of the light being reflected and scattered. This intern demonstrates the prior report’s finding that the band gap energy of greenly produced ZnO NPs has an inverse relationship with the average crystalline size [27].

3.3. Antibacterial Activity

S. aureus, B. subtilis, E. coli, and P. aeruginosa were all tested pathogens, and the ZnO NPs have exhibited a wide spectrum of antibacterial activity against all of them. This study evaluated how biomolecules and ZnO NPs worked together to combat four pathogenic diseases. Three concentrations of Co-clotrimazole, DMSO, and ZnO NPs (50, 75, and 100 μg/mL) were used to determine the zone of inhibition. The structural differences in bacteria cell walls are assumed to be the reason why ZnO NPs were discovered to have superior antibacterial action against Gram-positive bacteria than Gram-negative bacteria [5, 21]. The outer lipopolysaccharide membrane of Gram-negative bacteria, which renders the cell wall resistant to antibacterial chemical substances, is the source of these variances in morphological constitutions between these micro-organisms. Gram-positive bacteria, on the other hand, are more prone to permeability barriers since they only have an exterior peptidoglycan layer. Due to their more intricate cell walls than Gram-positive bacteria, Gram-negative bacteria are less susceptible to antibacterial medications and act as a diffusion barrier. The presence of bioactive chemicals that functions as capping and stabilizing agents on the surface of NPs can be partially blamed for the antibacterial activities of NPs. The ZnO NPs exhibited excellent antibacterial activity against both Gram-positive (S. aureus and B. subtilus) and Gram-negative (E. coli and P. aeruginosa) bacterial strains, as shown in Figure 7 (Table 3). When we compared the inhibition zone for each ratio of biosynthesized NPs, the maximum inhibition was seen for (2 : 3) ZnO NPs for each strain. This could be a result of the synergistic effects of phytochemicals included in plant extracts as well as the propensity of functional groups like phenolics and amines to interact with metal surfaces and disturb membrane integrity. At a sample concentration of 100 μg/mL, the inhibition zone for each strain is displayed in descending order for this maximum ratio; S. aureus (17.10 ± 0.45 mm), B. subtilus (16.10 ± 0.15 mm), E. coli (14.5 ± 0.32 mm), P. aeruginosa (13 ± 0.0 mm); the NPs with a 1 : 1 ratio showed lesser inhibition [24]. The alternative explanation could be that Zn antimicrobial activities are owing to its toxicity, which was noted in this experimental study and other similar works that have previously reported on the antimicrobial properties of this metal-based NPs [24, 28].

The inhibition zone of each strain was concentration-dependent, which is explained by the linear association between NPs concentration and the inhibitory effect, according to the table of antibacterial activity results. Scientists have proposed a few potential bactericidal pathways in response to ZnO NPs’ interactions with bacteria. Some researchers asserted that because smaller NPs have more surface reactivity, the released Zn²⁺ ions can penetrate them more deeply. One of the most popular theories for an antibacterial mechanism involves the release of Zn²⁺ from ZnO NPs, which is known to block several bacterial cell functions, including active transport, bacterial metabolism, and enzyme activity. Because of the toxicity properties of Zn²⁺, this ultimately causes bacteria to perish [28–32].

Some other researchers made the hypothesis that the production of reactive oxygen species (ROS), which results in oxidative stress and, ultimately, cell damage or death, is what triggers antibacterial action. ZnO NPs frequently use the generation of ROS as an antibacterial strategy [5, 24]. The attachment of NPs to the bacterial cell membrane by electrostatic forces is another potential mechanism for the antibacterial activity of ZnO NPs. This interaction could harm the bacterial cell’s integrity and alter the membrane plasma structure, allowing internal contents to seep out and ultimately leading to cell death [8, 21].

3.4. DPPH Radical Scavenging Assay

The synthesized ZnO NPs demonstrated the presence of antioxidant capabilities in an in vitro antioxidant experiment. The relationship between different volume ratios of synthetic ZnO NPs produced from aerial extracts and their capacity to scavenge DPPH radicals was proven (Table 4). All the antioxidant models showed a percentage of inhibitions, indicating that the produced ZnO NPs were effective at scavenging free radicals up to the specific concentration (Figure 8). According to the results in Table 4, the synthesized NPs have a negligible antioxidant effect in comparison to the reference standard.

P₃M₂ had the highest level of radical inhibition values of 83.67% at maximum concentration (100 μg/mL), which was comparable to L-ascorbic acid at this concentration when comparing the radical scavenging capability of various ratios. The P₂M₃ test yielded results that were second-most promising, with antiradical inhibition effects (100 μg/mL) of 72.79% compared with L-ascorbic acid, which inhibited at a rate of 98.59% at the same concentration (Table 4). Overall, the results demonstrate that the synthetic zinc oxide nanoparticles generated from R. multifidus aerial extracts offer a useful lead for future research into the development of antioxidant drugs to treat various cardiovascular diseases using more sophisticated instrumentation and cutting-edge technology. To compare the present result with previously reported work, zinc oxide-silver (ZnO–Ag) and zinc oxide-gold (ZnO–Au) nanocomposites were prepared through wet chemical process and laced into single-walled carbon nanotubes (SWCNTs) to yield ZnO–Ag-SWCNTs and ZnO–Au-SWCNTs hybrids, which shows significant phagocytic and bactericidal activities against the Gram-negative bacteria E. coli and Gram-positive S. aureus which support the current antibacterial effect of ZnO NPs using plant extracts [33].

4. Conclusion

Synthesis of ZnO NPs from aqueous extracts of aerial parts of R. multifidus using green methods was confirmed by the formation of yellow precipitates. The synthesized NPs were characterized by using XRD, SEM, TGA-DTA, FTIR, and UV-DRS, and the result is consistent with previously reported articles for ZnO NPs. FTIR result shows the nitro compounds and aromatic amines as reducing and capping agents, while analysis of produced ZnO nanoparticles revealed a UV–vis absorption peak at 442 nm. XRD examination demonstrated that every diffraction peak matched the hexagonal wurtzite structure of ZnO NPs. ZnO NPs’ spherical shape was confirmed by image taken using scanning electron microscopy. The synthesized ZnO NPs revealed a significant antibacterial activity against E. coli, P. aeruginosa, S. aureus, and B. subtilis relatively in all ratios. The significant phagocytic and bactericidal activities observed against the Gram-negative bacteria E. coli and Gram-positive S. aureus support the current antibacterial effect of ZnO NPs. The antiradical activities of produced ZnO NPs in various ratios show that plant material has the ability to scavenge free radicals. Generally, the present work indicates that there is a promising antibacterial and antioxidant nanoscale pharmaceutical product from R. multifidus aqueous aerial extracts using zinc metal and we strongly recommend testing more antifungal and anticancer activities of synthesized ZnO NPs in the future.

Data Availability

The data used to support the findings of this study are included within this article.

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Acknowledgments

The authors are grateful to Adama Science and Technology University for the support rendered towards this work. We are also grateful to the World Academy of Sciences (TWAS) and the United Nations Educational, Scientific and Cultural Organization (UNESCO) for financing this research with funds allocated to the AD research team under the TWAS Research Grant RGA No. 20-274 RG/CHE/AF/AC_G – FR3240314163.

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Copyright © 2023 Terfo Yilma et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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