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Article

Calotropis Gigantea Latex-Derived Zinc Oxide Nanoparticles: Biosynthesis, Characterization, and Biofunctional Applications

by
Jayalekshmi C
1,
Rajiv Periakaruppan
1,*,
Valentin Romanovski
2,*,
Karungan Selvaraj Vijai Selvaraj
3 and
Noura Al-Dayan
4
1
Department of Biotechnology, PSG College of Arts & Science, Coimbatore 641014, Tamil Nadu, India
2
Department of Materials Science and Engineering, University of Virginia, Charlottesville, VA 22904, USA
3
Vegetable Research Station, Tamil Nadu Agricultural University, Palur, Cuddalore 607102, Tamil Nadu, India
4
Department of Medical Lab Sciences, Prince Sattam Bin Abdulaziz University, Alkharj 16278, Saudi Arabia
*
Authors to whom correspondence should be addressed.
Eng 2024, 5(3), 1399-1406; https://doi.org/10.3390/eng5030073
Submission received: 3 June 2024 / Revised: 3 July 2024 / Accepted: 6 July 2024 / Published: 9 July 2024
(This article belongs to the Special Issue REPER Recent Materials Engineering Performances)
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Abstract

:
Latex of C. gigantea was used to synthesize zinc oxide nanoparticles (ZnO NPs) by the green chemistry approach. The crystalline size, shape, and purity of as-synthesized ZnO NPs were characterized through scanning electron microscopy with energy-dispersive X-ray spectroscopy, transmission electron microscopy, X-ray diffraction analysis, and Fourier-transform infrared spectroscopy techniques. Crystalline, spherical ZnO NPs with an average size of 21.8 nm were formed. In addition, the biological properties of the ZnO NPs, such as antioxidant and antibacterial activity, were evaluated by 2,2-diphenyl-1-picrylhydrazyl assay and the agar well-diffusion method. The highest free radical scavenging activities of 83.11 ± 1.89 % were observed at a concentration of 350 μg/mL of C. gigantea latex-mediated ZnO NPs. The latex in the C. gigantea latex-mediated ZnO NPs inhibited the growth of pathogenic bacteria. The maximum zone of inhibition was found in P. aeruginosa and S. aureus. C. gigantea latex-mediated ZnO NPs have significant biocompatibility and broad-spectrum antibacterial properties against wound-causing bacteria and, therefore, can be suggested for use in the formulation of novel creams or gels for healing applications.

1. Introduction

Nanotechnology is defined as advancement and exploration at the molecular or macromolecular, and even atomic, levels. Nanoparticles (NPs) are viewed as the fundamental components of nanotechnology and are described as particles with a size smaller than 100 nm in at least one dimension [1,2]. Various chemical and physical methods are currently being applied in the synthesis of nanoparticles [3,4]. Nevertheless, the biogenic reduction of metal precursors in aqueous media to generate NPs is environmentally friendly, cost-effective, and devoid of chemical impurities, making it ideal for biological and medical uses that require high purity. Biogenic reduction follows a “Bottom Up” approach similar to chemical reduction but utilizes natural product extracts as reducing agents with growth-controlling, built-in stabilizing, and capping capabilities [5].
Plant-produced nanoparticles exhibit greater stability and faster synthesis rates compared to those produced by microorganisms. Additionally, plant-derived nanoparticles display a wider range of shapes and sizes in contrast to those generated by other organisms. Researchers fascinated by the benefit of utilizing plant and plant-based materials for the biosynthesis of metal nanoparticles have investigated the mechanisms of bioreduction and metal ion absorption by plants, as well as the potential mechanisms involved in metal nanoparticle formation within plants [6]. Utilizing microorganisms poses a higher risk due to concerns regarding pathogenicity, in addition to the necessity of maintaining sizable cultures [7]. Different natural bio-resources such as seaweeds [8], leaves [9,10,11], flowers [12], bark [13], and microbes [14,15] have been utilized for the synthesis of zinc oxide nanoparticles.
Zinc oxide (ZnO) plays a crucial role as a component in numerous enzymes, sunscreens, and creams designed to alleviate pain and itching. The microcrystals of zinc oxide exhibit high efficiency in absorbing light within the UV-A and UV-B range of the spectrum, due to their wide band gap. The influence of zinc oxide on biological processes is contingent upon factors such as its structure, particle size, duration of exposure, concentration, pH levels, and compatibility with living organisms [16]. Despite the numerous reports indicating the significant antibacterial properties of CaO [17], FexOy [18], ZnO [19], HEA [20], and others [21,22,23] as a result of the production of Reactive Oxygen Species (ROS) on their surfaces, ZnO stands out as a particularly effective microorganism-resistant material. Moreover, ZnO possesses unique attributes such as structure-dependent, electrical, and thermal transport properties, making it both bio-safe and biocompatible [24].
Calotropis gigantea is otherwise called Milk Weed and Swallow Wort. Different compounds such as calotoxin, calotropin, uscharidin, gigantin, uscharin, cardiac glycosides, lupeol, and calactin are present in latex derived from C. gigantea [25]. The main objectives of this study were as follows: (i) investigate the biosynthesis of ZnO NPs using the aqueous mixed latex of C. gigantea and characterization of its size, shape, crystalline nature, and purity through spectroscopic and microscopic studies; (ii) investigate the antioxidant and antimicrobial properties of the as-synthesized ZnO NPs.

2. Materials and Methods

2.1. Materials and Reagents

C. gigantea latex was obtained from Nakshatra Garden at PSG College of Arts & Science, Coimbatore, 641014, India. Zinc nitrate and other chemicals (2,2-diphenyl-1-picrylhydrazyl, ethanol, methanol) with purity of 99.5% were obtained from Sigma Aldrich, Mumbai, India.

2.2. Synthesis of ZnO NPs Using C. gigantea Latex

An amount of 100 mL of 0.1 M of zinc nitrate solution was prepared. Next, 1% latex was added to the solution under constant stirring. The solution mixture was kept under vigorous stirring at 90 °C for 4–5 h. A pH level of 8 was maintained by a Benchtop pH-mV Meter (Sper Scientific, Scottsdale, AZ, USA). A white precipitate was achieved after this process and then centrifuged at 10,000 rpm for 20 min. The interaction of the 0.1 M zinc nitrate solution with 1% latex under constant stirring at 90 °C for 4–5 h and pH 8 involves the hydrolysis of zinc ions to form zinc hydroxide, which then interacts with latex particles to form a zinc–latex composite. Some zinc hydroxide may also convert to zinc oxide under these conditions. The process is driven by the elevated temperature, constant stirring, and basic pH, which facilitate the formation and stabilization of the composite material. The supernatant was removed and the white precipitate was collected. The precipitate was washed with ethanol and deionized water and then air-dried. Next, it was annealed at 400 °C for 1 h. Finally, the obtained white color particles were stored [26,27].

2.3. Characterization of ZnO NPs

The optical properties of ZnO NPs were determined by UV–visible spectroscopy (Shimadzu, Kyoto, Japan, UV–visible spectroscopy) at a wavelength range of 200–800 nm. X-ray diffraction (X’Pert Pro Panalytical) was employed to determine the phase and size of the ZnO NPs. FT-IR (Fourier-transform infrared spectroscopy) was used to analyze the functional groups of C. gigantea latex that were present as capping and stabilization agents of the ZnO NPs. The ZnO NPs were ground with KBr pellets (1% w/w), pressure was applied to form a disk, and then scanning was performed in the range of 400 to 4000 cm−1. Field-emission scanning electron microscopy (FESEM) analysis was conducted to determine the morphology of the ZnO NPs (Shimadzu miracle 10 models). Energy-dispersive X-ray (EDX) analysis provided details about the elemental composition of the zinc oxide nanoparticles.

2.4. Determination of Antibacterial Activity of ZnO NPs

Human pathogens such as Klebsiella pneumonia, Micrococcus sp., Proteases sp., Staphylococcus aureus, and Pseudomonas aeruginosa were acquired from the Microbiology Department of the PSG College of Arts & Science, Coimbatore, Tamil Nadu, India. The bacterial strains were grown in nutrient broth for antibacterial studies. The synthesized zinc oxide nanoparticles were tested for antibacterial activity against human pathogens by the agar well-diffusion method [28]. An amount of 100 µL of bacterial culture was prepared and swabbed on Mueller–Hinton agar plates.
The agar plates were punctured to create wells (5 mm) using a sterile gel puncture. Then, 100 µL of concentrations (25–100 μg/mL) of the synthesized zinc oxide nanoparticles were added to the wells. An amount of 100 µL (10 μg/mL) of positive control (Tetracycline) was prepared and added to the wells of all plates using micropipettes and incubated for 48 h at room temperature. The zone of inhibition was measured in millimeters. Each assay was carried out with five replicates and mean values were recorded [29].

2.5. Determination of Antioxidant Activity of ZnO NPs

According to the study by Periakaruppan et al. [30], the antioxidant properties of the zinc oxide nanoparticles were determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Different concentrations (10, 50, 150, 250, and 350 μg/mL) of ZnO NPs were prepared and ascorbic acid was used as a standard solution and transferred to different test tubes. The reference was 2 mL of methanol. DPPH 0.3 mM was freshly prepared (1 mL) and added to different tubes and the solution was incubated for 30 min in a dark condition. The absorbance peak was measured at 517 nm and methanol was used as a blank solution. The DPPH activity of the ZnO NPs was calculated using Formula (1):
Aa = (Ao − Ai/Ao) · 100%,
where Aa is the percentage (%) of the DPPH radical scavenging effect; Ai is the average absorbance of the tested solution; and Ao is the average absorbance of the DPPH solution.

3. Results and Discussion

3.1. Characterization of C. gigantea Latex-Mediated ZnO NPs

The UV-Vis spectrum determined that C. gigantea latex-mediated ZnO NPs show a strong absorption peak at 346 nm and thus ensure the formation of ZnO NPs (Figure 1a). A band gap of 3.37 eV was observed for C. gigantea latex-mediated ZnO NPs. The optical properties of the green-synthesized ZnO NPs were assessed using UV-Vis spectroscopy [31,32]. Our optical property findings are in good agreement with the reported values in the literature [33,34,35].
Figure 1b represents the FTIR spectra of C. gigantea latex-mediated ZnO NPs. The functional groups of the C. gigantea latex-mediated ZnO NPs were formed due to the interaction of biomolecules or secondary metabolites from latex, which were responsible for the reduction of zinc ions to ZnO NPs. Peaks at 493, 555, 601, 694, and 879 cm−1 were observed in the FTIR spectra and refer to the occurrence of metal oxide groups, which clearly reveals the formation of ZnO NPs. A peak at 1635 cm−1 was obtained because of vibrations of hydroxyl groups from surface-adsorbed water. The band at 1334 cm−1 indicates the presence of C–N stretching bonds. C–O stretching was detected by the existence of a peak at 1126 cm−1. Moreover, bands were detected at 1419 cm−1 and 1573 cm−1, which corresponded to C-H bonding. Similarly, Geetha et al. [36] synthesized ZnO NPs using Euphorbia Jatropa latex as a reducing agent and reported the various functional groups, namely ZnO bonding, H-bonded, and O–H groups.
The XRD patterns of C. gigantea latex-mediated ZnO NPs are shown in Figure 1c. The XRD diffractogram exhibits different diffraction peaks at the (100), (002), (101), (102), (110), (103), (112), and (201) reflection lines that refer to the hexagonal crystal system of the P63mc space group of the ZnO phase. The narrow and strong peaks denote that the C. gigantea latex-mediated ZnO NPs were crystalline in nature. The average size of the C. gigantea latex-mediated ZnO NPs was determined by Debye Scherrer’s formula (D = Kλ/βCosθ). The average size of the C. gigantea latex-mediated ZnO crystallites was around 21.8 nm, which was calculated for the main peak at 36.55 2theta (hkl 011) (JCPDS card number 36-1451). Miri et al. [37] biosynthesized zinc oxide nanoparticles and investigated their crystalline nature. They determined the average size of particles using XRD analysis. The ZnO NP size range according to the SEM images was about 50–200 nm. This means that the obtained ZnO NPs were polycrystalline in nature. The green chemistry approach by Vanathi et al. [27] was used for phyto-mediated zinc oxide nanoparticle synthesis and investigation of the crystalline nature of the NPs was studied by XRD analysis.
Figure 2 shows the shape of C. gigantea latex-mediated ZnO NPs. Under various magnifications, the shape of the C. gigantea latex-mediated ZnO NPs was examined and we confirmed that particles were spherical with a few agglomerated forms. Al-darwesh et al. [38] synthesized Ficus carica latex-mediated zinc oxide nanoparticles and observed a spherical shape with an average size of 29.3 nm using a scanning electron microscope. E. átirucalli plant latex-mediated ZnO nanoparticles were fabricated by Kumar et al. [39] who reported the formation of spherically shaped particles with agglomeration.
The elemental composition and formation of ZnO NPs were investigated by an energy-dispersive X-ray (EDX) diffractive study. The EDX spectra confirm the presence of oxygen (47.81 at%), zinc (40.80 at%), and carbon (11.39 at%) (Figure 2a) signals of ZnO NPs and prove the formation of ZnO NPs. Similar results were derived for Mangifera indica leaf-mediated ZnO NPs in EDX spectra by Rajeshkumar et al. [40] who observed a pure form of zinc oxide nanoparticles. Rad et al. [10] proved the elemental composition of Mentha pulegium-mediated ZnO NPs by EDX analysis.

3.2. Analysis of Antioxidant Activity

The antioxidant potential of C. gigantea latex-mediated ZnO NPs was studied using DPPH assay (Table 1). Ascorbic acid was used as a standard solution. The DPPH assay results demonstrate that the antioxidant properties increased with increasing concentrations of C. gigantea latex-mediated ZnO NPs (10–350 μg/mL) and the % of free radical scavenging activity was boosted. The highest free radical scavenging activity (83.11 ± 1.89%) was observed at a concentration of 350 μg/mL of C. gigantea latex-mediated ZnO NPs. The lowest free radical scavenging activity was detected at a concentration of 10 μg/mL of C. gigantea latex-mediated ZnO NPs. Abdelhakim et al. [41] reported that biogenic ZnO NPs show promising antioxidant potential with IC50 of 102 µg/mL. The in vitro antioxidant activity of phyto-assisted ZnO NPs was determined by Rehana et al. [42] who found significant free radical scavenging activity.

3.3. Antibacterial Activity

C. gigantea latex-assisted ZnO NPs show significant antibacterial activity against wound-causing bacteria such as Proteases sp., Klebsiella pneumonia, Micrococcus sp., Staphylococcus aureus, and Pseudomonas aeruginosa (Figure 3). The maximum zone of inhibition was observed against S. aureus and P. aeruginosa at various concentrations of C. gigantea latex-mediated ZnO NPs. Al-darwesh et al. [38] stated that the biological activity and antibacterial properties of F. carica latex-mediated ZnO NPs kill and reduce the growth and activity of S. aureus, K. pneumoniae, B. subtilis, E. coli, and P. aeruginosa. Biogenic ZnO NPs were synthesized using different plant extracts and exhibited similar antibacterial properties [43,44,45,46].

4. Conclusions

C. gigantea latex-mediated ZnO NPs were synthesized by rapid, simple, and eco-friendly methods. An aqueous-based latex extract was employed to synthesize the ZnO NPs. The formation of C. gigantea latex-mediated ZnO NPs was confirmed and physicochemical properties were characterized through various microscopic and spectroscopic methods. The synthesized nanoparticles were well crystalline in nature and the average size was determined to be 21.8 nm. C. gigantea latex-mediated ZnO NPs with antioxidant potential were successfully synthesized and proved by DPPH assay. The antibacterial activity of C. gigantea latex-mediated ZnO NPs was assessed using wound-causing bacteria. The maximum zone of inhibition was observed against P. aeruginosa and S. aureus. The results conclude that the as-synthesized C. gigantea latex-mediated ZnO NPs have excellent biocompatibility and broad-spectrum antibacterial activity against wound-causing bacteria. In summary, C. gigantea latex-mediated ZnO NPs can be suggested for use in the formulation of new creams or gels for therapeutic applications.

Author Contributions

J.C.: investigation, writing—original draft preparation, project administration; R.P.: conceptualization, supervision, writing—review and editing; V.R.: formal analysis, validation, data curation, investigation, writing—review and editing. K.S.V.S.: formal analysis, validation, data curation; N.A.-D.: formal analysis, validation, data curation. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

All data, models, and code generated or used during this study appear in the submitted article.

Conflicts of Interest

The authors declare no conflicts of interest.

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Figure 1. UV-Vis spectrum of C. gigantea latex-mediated ZnO NPs (a), FTIR analysis of C. gigantea latex-mediated ZnO NPs (b), and XRD analysis of C. gigantea latex-mediated ZnO NPs (c).
Figure 1. UV-Vis spectrum of C. gigantea latex-mediated ZnO NPs (a), FTIR analysis of C. gigantea latex-mediated ZnO NPs (b), and XRD analysis of C. gigantea latex-mediated ZnO NPs (c).
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Figure 2. FESEM (a) and TEM (b,c) images of C. gigantea latex-mediated ZnO NPs.
Figure 2. FESEM (a) and TEM (b,c) images of C. gigantea latex-mediated ZnO NPs.
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Figure 3. Antibacterial activity of C. gigantea latex-mediated ZnO NPs.
Figure 3. Antibacterial activity of C. gigantea latex-mediated ZnO NPs.
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Table 1. Antioxidant analysis of C. gigantea latex-mediated ZnO NPs.
Table 1. Antioxidant analysis of C. gigantea latex-mediated ZnO NPs.
Concentration (µg/mL)% of InhibitionIC50 (µg/mL)
1058.20 ± 1.2912.52
5065.57 ± 1.58
15068.85 ± 1.47
25077.21 ± 1.36
35083.11 ± 1.89
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C, J.; Periakaruppan, R.; Romanovski, V.; Vijai Selvaraj, K.S.; Al-Dayan, N. Calotropis Gigantea Latex-Derived Zinc Oxide Nanoparticles: Biosynthesis, Characterization, and Biofunctional Applications. Eng 2024, 5, 1399-1406. https://doi.org/10.3390/eng5030073

AMA Style

C J, Periakaruppan R, Romanovski V, Vijai Selvaraj KS, Al-Dayan N. Calotropis Gigantea Latex-Derived Zinc Oxide Nanoparticles: Biosynthesis, Characterization, and Biofunctional Applications. Eng. 2024; 5(3):1399-1406. https://doi.org/10.3390/eng5030073

Chicago/Turabian Style

C, Jayalekshmi, Rajiv Periakaruppan, Valentin Romanovski, Karungan Selvaraj Vijai Selvaraj, and Noura Al-Dayan. 2024. "Calotropis Gigantea Latex-Derived Zinc Oxide Nanoparticles: Biosynthesis, Characterization, and Biofunctional Applications" Eng 5, no. 3: 1399-1406. https://doi.org/10.3390/eng5030073

APA Style

C, J., Periakaruppan, R., Romanovski, V., Vijai Selvaraj, K. S., & Al-Dayan, N. (2024). Calotropis Gigantea Latex-Derived Zinc Oxide Nanoparticles: Biosynthesis, Characterization, and Biofunctional Applications. Eng, 5(3), 1399-1406. https://doi.org/10.3390/eng5030073

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