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Therapeutic potential of sustainable zinc oxide nanoparticles biosynthesized using Tradescantia spathacea aqueous leaf extract

  • Sumreen Sultana , Bagepalli Shivaram Ashwini , Umme Hani , Nazima Haider , Mohammad Nasser Alomary , Venkateshppa Bhavana , Bangari Daruka Prasad , Tekupalli Ravikiran , Mohammad Azam Ansari EMAIL logo and Thimappa Ramachandrappa Lakshmeesha EMAIL logo
Published/Copyright: June 10, 2025
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Green Processing and Synthesis
From the journal Green Processing and Synthesis Volume 14 Issue 1

Abstract

The present study focused on the synthesis of zinc oxide nanoparticles (ZnO NPs) utilizing an aqueous extract (leaves) of Tradescantia spathacea and assessed their antibacterial and anticancer activities. The characterization of NPs was performed using XRD, UV-Vis spectroscopy, Fourier-transform infrared, scanning electron microscopy-energy-dispersive X-ray analysis, and HRTEM with selected area electron diffraction. The antibacterial activity of ZnO NPs was evaluated using the disc diffusion method and the trypan blue dye exclusion method. The anticancer effects in HeLa cells were assessed using the MTT assay, while cellular uptake was assessed through Rhodamine B isothiocyanate-labeled ZnO NPs. The cytotoxic properties of NPs were assessed by estimating the mitochondrial membrane potential (MMP) and apoptosis by Hoechst, propidium iodide, Annexin V-FITC staining, and cell cycle distribution. The synthesized NPs exhibited antibacterial ability with the highest inhibition zone measuring 13.2 mm at a concentration of 1 mg·mL−1. The MTT assay on HeLa cells showed dose-dependent viability ranging from 88% to 24%, with an IC50 value of 84.26 μg·mL−1. JC-1 and Hoechst staining assays confirmed the impairment of MMP and apoptosis, with significant cell cycle arrest observed in the Sub G0/G1, S, and G2/M phases, indicating a disruption in the regular cell cycle. In conclusion, green-synthesized ZnO NPs displayed significant antibacterial and anticancer properties, emphasizing their potential for use in biomedicine and healthcare applications.

Keywords: HeLa cells; Escherichia coli ; green synthesis; mitochondrial membrane potential; apoptosis; cell cycle

1 Introduction

Cancer is one of the main reasons for mortality across the globe, with nearly 20 million new cases reported in 2022, and this number is further expected to increase to 28.4 million by 2040 [1,2]. Among various types of cancers in women, cervical cancer ranks as the fourth most prevalent cause of death. Almost 99% of the cervical cancer cases are due to persistent infection with human papillomavirus [3]. Currently, available drugs for curing cancer lack potency and specificity and have high side effects. Insights into the molecular mechanisms underlying cervical cancer have sparked interest in developing novel treatment strategies as an alternative to conventional medications, including chemotherapy and radiation [4]. Furthermore, cancer patients are also highly vulnerable to a wide range of bacterial infections, which is a great obstacle because of the direct and indirect effects on the immune systems [5]. They are especially at risk due to surgical issues, chemotherapy, radiotherapy, and immunosuppressive medications [6].

The World Health Organization claims that antimicrobial resistance poses a serious global health threat, contributing to 4.95 million deaths overall [7,8]. The overuse and misuse of antimicrobials in humans, animals, and plants are key factors in the emergence of drug-resistant pathogens [9]. Particularly, Escherichia coli is a prevalent pathogen associated with cervical cancers, and higher levels of IL-10 in cervical tissues suggest that E. coli may contribute to cancer development [10]. In cancer patients, antibiotic failure enhances the risk of sepsis and sepsis-related mortality [11]. Consequently, it becomes vital to investigate the development of safe, efficient, and eco-friendly materials with both anticancer and antibacterial properties.

Recently, nanomaterials have garnered significant interest in various sectors because of their unique intrinsic physicochemical characteristics [12]. Globally, nanoparticles (NPs) are becoming a vital option for the treatment of infections resistant to drugs as well as for selective targetivity to cancerous cells [13]. Zinc oxide NPs (ZnO NPs) are the metal oxide NPs that are recognized by the US Food and Drug Administration as “generally recognized as safe (GRAS)” for human and animal consumption [14]. Moreover, these NPs generate reactive oxygen species (ROS), which are harmful to microorganisms under light and further exhibit antimicrobial effects in the dark through cell membrane damage and Zn²⁺ ion release. ZnO NPs are also known to elevate ROS generation and induce apoptotic cell death in cancer cells. These attributes make ZnO NPs potent microbicidal and anticancer agents [15,16].

ZnO NPs can be efficiently prepared through green synthesis, offering benefits over chemical and physical approaches with regard to production expenses and the impact on the environment. While the methods involving physicochemical production of NPs have been frequently employed, biogenic synthesis is the best choice due to its low environmental impact [17]. This approach allows for rapid and scalable production while reducing the use of harmful chemicals that can pose toxicity and health risks [18]. Medicinal plants serve as a rich source of phytochemicals suitable for treating a variety of disorders, and they can also serve as potential sources for new drug development. These plant products continue to be the most readily available and reasonably priced medications for basic healthcare in poor nations [19]. They contain numerous phytochemicals like alkaloids, tannins, ascorbic and carboxylic acids, amides, aldehydes, ketones, flavonoids, and phenols that serve as stabilizing and reducing agents, facilitating the reaction with metallic salts to produce nanosized materials. Additionally, these phytochemicals contribute to the stability, biocompatibility, and eco-friendliness of NPs [20].

Tradescantia spathacea, commonly referred to as “oyster plant,” is a part of the Commelinaceae family and possesses several therapeutic properties, including anti-diabetic effects, cosmetic benefits, anti-inflammatory properties, wound-healing potential, and applications in the treatment of venereal diseases, hematemesis, dysentery, asthma, intestinal infections, hemorrhage, antimicrobial activity, allergic rhinitis, detoxification, antioxidant activity, and neuroprotection [21,22,23]. In India, it is extensively used in folk medicine to cure a wide range of ailments, cancer being the chief among them [24]. Even though extensive studies are available on the biosynthesis of ZnO NPs using plant extracts, limited literature is available regarding the utilization of T. spathacea for investigating the antibacterial and anticancer activities. Recently, a study reported the use of T. spathacea for developing an electrochemical sensor based on a ZnO and multi-walled carbon nanotube nanocomposite for chloramphenicol detection in food samples [24]. Another study reported the synthesis of tin oxide NPs using aqueous leaf extract of T. spathacea for studying the photoantioxidant activity [25]. Therefore, the novelty of our study lies in exploring the antibacterial activity of ZnO NPs synthesized using T. spathacea against E. coli isolated from sputum, as well as exploring its anticancer effect against HeLa cells.

2 Materials and methods

2.1 Preparation of T. spathacea aqueous leaf extract

Fresh T. spathacea leaves were gathered from the Department of Microbiology and Biotechnology at Bangalore University in Bengaluru. The precise collection site, along with the plant material, was mapped using ArcGIS (version 9.3), which is depicted in Figure 1. The collected plant sample was authenticated (RRCBI-mus587) as T. spathacea by Dr V. Rama Rao from the Central Ayurveda Research Institute. The aqueous extract of leaves was prepared by carefully selecting disease-free samples and, thoroughly rinsing under running tap water several times and subsequently washing with double-distilled water to ensure cleanliness. After this, the leaves were soaked in double-distilled water for 1 h. Following the soaking process, the leaves were finely blended using an electric blender and centrifuged (3,000 rpm, 5 min). The supernatant obtained from this step was utilized in synthesizing the ZnO NPs [26].

Figure 1 (a) T. spathacea plant, and (b) sampling location of T. spathacea.
Figure 1

(a) T. spathacea plant, and (b) sampling location of T. spathacea.

2.2 Qualitative phytochemical analysis of the aqueous extract of T. spathacea leaves

The preliminary phytochemical analysis of the prepared leaf extract was accomplished to detect various bioactive components such as carbohydrates (Molisch’s test), proteins, tannins, cardiac glycosides (Keller–Killiani test), steroids, flavonoids (alkaline reagent test), and alkaloids (Dragendorff’s test) according to the standard protocols [27,28].

2.3 Gas chromatography-mass spectrometry (GC-MS) analysis

The T. spathacea leaf extract was analyzed using a Clarus 680 gas chromatograph equipped with an Elite-5MS column (30 m × 0.25 mm ID × 250 µm film thickness, 5% biphenyl, 95% dimethylpolysiloxane). The carrier gas (helium) at a flow rate of 1 mL·min−1 was used. The sample (1 µL) was inserted at an injector temperature of 260°C. The temperature of the oven was initially adjusted to 60°C for 2 min, then increased up to 300°C at 10 °C·min−1, and maintained for 6 min. Mass detection was performed using electron impact ionization at 70 eV that scanned fragment masses from 40 to 600 Da. Compounds were identified using the NIST (2008) GC-MS library.

2.4 Synthesis of ZnO NPs from the aqueous extract of T. spathacea leaves

The synthesis of ZnO NPs was accomplished by solution combustion [29]. Zinc nitrate hexahydrate [Zn(NO3)2·6H2O] (2.5 g) was mixed with the T. spathacea filtrate (20 mL) and subjected to magnetic stirring for 15 min. This solution was then placed into a crucible and moved to a pre-heated muffle furnace, which was maintained for 2 h at 400 ± 10°C. After combustion, the material was collected for further characterization. Besides, the zinc oxide without plant extract (control) was also synthesized under the same conditions in order to provide emphasis on the role of the plant extract in the synthesis of ZnO NPs.

2.5 Characterization of ZnO NPs

The formation of ZnO NPs was confirmed using various analytical techniques. A UV-Vis spectrophotometer (Shimadzu 1800, Japan) was used for obtaining absorption spectra from 200 to 800 nm, confirming their formation. Fourier-transform infrared (FT-IR) spectroscopy (Agilent Technologies) was utilized for recording spectra from 4,000 to 400 cm⁻¹ to identify the functional groups. The crystalline nature and phase purity were analyzed via powder X-ray diffraction (Shimadzu XRD 7000 maxima, Japan) over a range of 20–80° with Cu Kα radiation (1.5418 Å). The elemental composition and surface morphology were assessed by energy-dispersive X-ray (EDX) analysis and scanning electron microscopy (SEM; Hitachi TM-3000), respectively. High-resolution transmission electron microscopy (HR-TEM) analysis (Tecnai G2, F30 FEI) was performed to determine the NP size and provided selected area electron diffraction (SAED) patterns. Together, these techniques offered a comprehensive characterization of the synthesized ZnO NPs, including their size, structure, composition, and morphology.

2.6 Isolation and identification of E. coli

E. coli was isolated from a fresh sample (sputum) by mixing it at a ratio of 1:1 with phosphate-buffered saline (PBS). A 1 mL aliquot of this diluted sputum was inoculated onto blood agar and MacConkey agar plates and incubated (37°C, 24 h). Colonies were chosen based on the preliminary morphology and colony characteristics, which were then transferred to nutrient agar plates, and molecular identification was conducted using the ITS region of the 16S rRNA gene. The obtained sequence was deposited in the GenBank database (NCBI), followed by a BLAST homology search [30].

2.7 Antibacterial efficacy of ZnO NPs

The antibacterial efficiency of ZnO NPs against isolated E. coli was evaluated using the disc diffusion method. An E. coli suspension was cultured for 24 h in Mueller–Hinton broth and adjusted to the McFarland standard of 0.5, corresponding to approximately 1.5 × 108 CFU·mL−1. Approximately 100 µL of this suspension was spread on Mueller–Hinton agar plates. Sterile discs were loaded with varying concentrations (1, 0.5, and 0.25 mg·mL−1) of ZnO NPs along with a positive control streptomycin (1 μg·mL−1) and sterile double-distilled water as a negative control. After placing the discs on the inoculated plates, they were maintained for 24 h at 37°C. The inhibition was assessed by the measurement of the clear zones surrounding the discs [31].

2.8 Estimation of ROS generation

ROS generation was assessed using 2′,7′-dichlorodihydrofluorescein diacetate (DCFHDA). Bacterial cell (E. coli) suspensions treated with ZnO NPs (1, 0.5, and 0.25 mg·mL−1) were centrifuged at 300 g for 30 min at 4°C. Hydrogen peroxide (H2O2, 0.8 mM) was used as a positive control, while the supernatant from non-treated cells served as a negative control. The supernatant from all the groups was incubated with 100 μM DCFHDA in darkness for 1 h. The fluorescence intensity was measured using a multimode reader (Biotek, Synergy LS) equipped with a green filter with an excitation wavelength of 485 nm and an emission wavelength of 530 nm [32].

2.9 Trypan blue dye exclusion assay

The dye exclusion (trypan blue) technique was used to differentiate between live and non-viable bacteria. ZnO NPs (1 mg·mL−1) were treated with the bacterial cell suspension (1.5 × 108 CFU·mL−1) and incubated at 37°C (24 h). This suspension was then subjected to centrifugation, and the pellet obtained was then combined with a 0.4% solution of trypan blue. A 10 µL aliquot of this suspension was positioned on a glass slide and visualized under a bright-field microscope (LM521710, Lynx) at 40× magnification. This method enabled clear differentiation between live and dead bacterial cells, facilitating the evaluation of the antimicrobial effects of ZnO NPs on cell viability and their potential applications [33].

3 In vitro anticancer activity

3.1 Cell culture and maintenance

HeLa cells were provided by the National Centre for Cell Science (Pune, India) and cultivated in DMEM containing FBS (10%) and 1% antimycotic–antibiotic solution. These cells were kept in a CO₂ incubator set at 5% CO₂ and 37°C. The culture medium was refreshed every 2 days and subcultured once the cells became 70–80% confluent.

3.2 Determination of cell viability by MTT assay

The method of Mosmann was followed for performing the MTT assay [34]. A 96-well plate was seeded with HeLa cells (1 × 10⁴ cells per well) and allowed to attach for 24 h. The cells were then exposed to varying concentrations of ZnO NPs (10–320 μg·mL−1) along with cisplatin (10 μg·mL−1) as a standard reference and incubated for an additional 24 h. To each well, 10 μL of MTT solution prepared in PBS (5 mg·mL−1) was added and kept in the dark (4 h, 37°C). Then, 100 μL of DMSO was added after the removal of the medium to dissolve the formazan crystals. A microplate reader (Biotek 800 TS, Agilent Technologies) was used for determining the absorbance at 570 nm. The cell viability (%) was determined using the following equation:

Cell viability ( % ) = Absorbance ( Sample ) Absorbance ( Control ) × 100 (1)

3.3 Cellular uptake of ZnO NPs

3.3.1 Preparation of rhodamine B isothiocyanate (RITC)-labelled ZnO NPs

The RITC-labelled ZnO NPs were prepared according to the previously published protocol [35]. RITC-tagged ZnO NPs were synthesized in a two-step process. First, ZnO NPs were converted into amine-functionalized NPs, and then RITC was conjugated to amine-functionalized ZnO NPs. Initially, ZnO NPs (0.5 g) were dissolved in DMSO (50 mL) and left in a bath sonicator for around 1 h. The resulting dispersion was transferred to a round-bottom flask connected to a reflux condenser. Then, 400 µL of 3-aminopropyl triethoxy silane (APTES) was added to the mixture and refluxed for 3 h at 120°C followed by centrifugation (12,000 rpm, 15 min). The unreacted APTES was removed by repeated washing with ethanol and further dried overnight at 60°C. After this, the dried powder of amine-functionalized ZnO NPs dissolved in 0.1 M NaHCO3 was mixed with RITC dissolved in DMSO (0.5 mg·mL−1) in an equal ratio and stirred at room temperature for 24 h in the dark for the formation of RITC-labelled ZnO NPs.

3.3.2 Cellular internalization of ZnO NPs

The qualitative cellular uptake was investigated using confocal laser scanning microscopy [36]. HeLa cells (4 × 104 cells) were allowed to grow on a coverslip for 24 h. Then, the sub-toxic concentration of RITC-labelled ZnO NPs was incubated for 24 h. Subsequently, the PBS-washed cells were stained with Hoechst (1 μg·mL−1) for 20 min (dark). The reddish fluorescence of RITC-labelled ZnO NPs was detected at their respective excitation and emission wavelengths. Subsequently, the ImageJ software was used to calculate the mean fluorescence intensity of RITC staining in the cytoplasm.

3.4 Assessment of mitochondrial membrane potential (MMP)

The MMP was measured using a fluorescent stain, JC-1, according to the previously published protocol [37]. The HeLa cell seeding was achieved as described earlier and further exposed to ZnO NPs for 24 h. JC-1 (10 μM) was then added and left in the dark (30 min, 37°C). The cells were gently washed with PBS and resuspended in fresh PBS, followed by flow cytometric analysis.

3.5 Apoptotic analysis

3.5.1 Hoechst staining

Hoechst staining was performed following the procedure of Lee et al. [38]. The cells were grown on confocal dishes for 24 h and subsequently treated with ZnO NPs. After 24 h, the cells were washed with PBS and stained with Hoechst 33432 dissolved in PBS (1 μg·mL−1) for 20 min. A confocal microscope (CKX41, Olympus Corporation, Japan) was used for examining the stained cells. The condensed and damaged nuclei with bright glowing appearance were considered as an indicator of apoptotic cells.

3.5.2 Detection of apoptosis using Annexin V-FITC and PI

Apoptosis was performed according to the manufacturer’s instructions (Elab Sciences). Cells were seeded and left for 24 h and exposed to ZnO NPs for another 24 h. Then, 200 μL of the assay buffer was added, and the cells were processed after rinsing thrice with cold PBS. Subsequently, 10 μL of annexin V was mixed with 500 μL of the cell suspension and left in the dark for 10 min, followed by mixing 5 μL of PI. A flow cytometer was used for apoptotic analysis.

3.6 Cell cycle analysis

PI staining was used for the assessment of cell cycle distribution following the earlier protocol [39]. The cells were processed in a 6-well plate, as described earlier. The trypsinized cells were centrifuged, and the pellet was mixed with 70% ethanol for fixation and left overnight at 4℃. The PI staining solution, comprised of 50 μg·mL−1 PI, 0.05% Triton X-100, and 0.1 mg·mL−1 RNase A, was mixed and left for 40 min at 37°C followed by flow cytometric analysis.

3.7 Statistical analysis

GraphPad prism was used for performing the statistical analysis. Data were presented as (n = 3) mean ± SE, followed by one-way analysis of variance (ANOVA). Post hoc analysis was done through Tukey’s test. Probability values less than 0.05 were considered to be significant.

4 Results and discussion

4.1 Phytochemical analysis

Traditional herbs are an abundant source of phytochemicals, including alkaloids, flavonoids, terpenoids, and tannins, which act as secondary metabolites with therapeutic effects against diseases such as antibiotic-resistant infections and cancer. This study highlights their potential for effective treatments with minimal side effects [40]. Moreover, in NP synthesis, plant-derived phytochemicals play a crucial role in reducing and stabilizing metal ions, facilitating NP formation, and preventing agglomeration [41]. The results of the phytochemical analysis of the aqueous extracts from T. spathacea leaves are presented in Table 1. This analysis indicates the prevalence of carbohydrates, steroids, saponins, flavonoids, and tannins in the leaf extract. Supporting our findings, reports of Pulipaka et al. [42] demonstrated that the T. spathacea aqueous extract contains carbohydrates, proteins, saponins, tannins, terpenoids, flavonoids, alkaloids, resins, and coumarins.

Table 1

Phytochemical analysis of aqueous leaf extract of T. spathacea

Sl. no. Phytochemical test Results
1 Carbohydrates +
2 Tannin −
3 Steroid +
4 Saponin +
5 Flavonoid +
6 Alkaloid +
7 Glycosides −
8 Proteins +

4.2 GC-MS analysis

Plants are the reservoirs of several bioactive compounds and are increasingly being used to discover novel drugs with better therapeutic efficacy. GC-MS has been recognized as a reliable technique for the detection and identification of bioactive compounds [43]. In our study, 13 compounds were identified based on their retention time and molecular formula, as shown in Figure 2 and Table 2. The compounds that were detected include 3-amino-2-oxazolidinone, cyclopentane, 1-(2-decyldodecyl)-2,4-dimethyl, palmitic acid vinyl ester, 3-methyl-2-(2-oxopropyl)furan, 2-isopropyl-5-methylcyclohexyl 3-(1-phenyl-3-oxobutyl)-coumarin-4-yl carbonate, 9-octadecenoic acid (z)-octadecyl ester, cyclopentane, 1-(2-decyldodecyl)-2,4-dimethyl, 2-isopropyl-5-methylcyclohexyl 3-(1-phenyl-3-oxobutyl)-coumarin-4-yl carbonate, cyclopentane, 1-(2-decyldodecyl)-2,4-dimethyl, decyl oleate, 1-hexyl-2-nitrocyclohexane, and 3-methyl-2-(2-oxopropyl)furan, I-propyl 9-octadecenoate.

Figure 2 GC-MS chromatogram of T. spathacea leaf aqueous extract.
Figure 2

GC-MS chromatogram of T. spathacea leaf aqueous extract.

Table 2

Phytoconstituents of aqueous leaf extract of T. spathacea identified by GC-MS

Peak no. Components Retention time (min) Molecular weight (g·mol−1) Molecular formula
1 3-Amino-2-oxazolidinone 1.34 102 C3H6O2N2
2 Cyclopentane, 1-(2-decyldodecyl)-2,4-dimethyl- 6.31 406 C29H58
3 Palmitic acid vinyl ester 7.26 282 C18H34O2
4 Palmitic acid vinyl ester 8.84 282 C18H34O2
5 3-Methyl-2-(2-oxopropyl)furan 10.21 138 C8H10O2
6 2-Isopropyl-5-methylcyclohexyl 3-(1-phenyl-3-oxobutyl)-coumarin-4-yl carbonate 10.89 490 C30H34O6
7 9-Octadecenoic acid (z)-, octadecyl ester 13.37 534 C36H70O2
8 Cyclopentane, 1-(2-decyldodecyl)-2,4-dimethyl- 13.62 406 C29H58
9 2-Isopropyl-5-methylcyclohexyl 3-(1-phenyl-3-oxobutyl)-coumarin-4-yl carbonate 15.65 490 C30H34O6
10 Cyclopentane, 1-(2-decyldodecyl)-2,4-dimethyl- 17.93 406 C29H58
11 9-Octadecenoic acid (z)-, octadecyl ester 18.45 534 C36H70O2
12 Decyl oleate 19.90 422 C28H54O2
13 1-Hexyl-2-nitrocyclohexane 22.68 213 C12H23O2N
14 3-Methyl-2-(2-oxopropyl)furan 25.06 138 C8H10O2
15 1-Propyl 9-octadecenoate 26.17 324 C21H40O2

4.3 Characterization of ZnO NPs

The formation of ZnO NPs synthesized with T. spathacea extract and the as-formed bulk zinc oxide (ZnO) without plant extract, along with its UV-Vis absorption and FTIR spectra, are shown in Figures S1 and S2 of the supplementary file. The synthesized ZnO NPs were characterized using techniques like UV-Vis, FTIR, and XRD. The UV-Vis spectrum of T. spathacea synthesized ZnO NPs displays a strong absorption peak at approximately 288 nm, indicating their maximum surface plasmon resonance, as shown in Figure 3a. This peak confirms the successful synthesis of ZnO NPs. This observation is similar to the findings by Aliyu et al. [44], which recorded an absorption peak at 281 nm for ZnO NPs synthesized with Ziziphus spinachristi extracts. Slight differences in absorption wavelengths resulted from variations in synthesis techniques and the distinct phytochemicals present in the specific plant parts [45].

Figure 3 (a) UV-Vis, (b) FT-IR, and (c) XRD spectra of ZnO NPs.
Figure 3

(a) UV-Vis, (b) FT-IR, and (c) XRD spectra of ZnO NPs.

FTIR measurements were conducted to analyze the functional groups in ZnO NPs synthesized from T. spathacea leaf extract. As shown in Figure 3b, the FTIR spectra revealed significant peaks: an absorption peak at 3,322 cm⁻¹ corresponding to –OH stretching, a peak at 1,630 cm⁻¹ corresponding to –C═C– stretching and a peak at 522 cm⁻¹, indicating the metal oxide (Zn–O) stretching vibration mode. Our results are in accordance with those of Hemanth Kumar et al. [46], who also detected similar peaks in ZnO NPs derived from Simarouba glauca DC, suggesting that functional groups play a crucial role in the stabilization and synthesis of ZnO NPs from plant extracts.

The powder XRD pattern of the synthesized ZnO NPs is presented in Figure 3c. All the observed diffraction peaks match the (100), (002), (101), (102), (110), (103), (200), (112), (201), (004), and (202) planes, indexed as per the JCPDS Card No. 36-1451 [47]. According to established references, every diffraction peak was properly attributed to the hexagonal phase of ZnO NPs. The (100), (002), and (101) planes relate to the Bragg angles of 31°, 34°, and 36°, respectively, indicating the good crystallinity of ZnO NPs. The XRD analysis revealed that the (101) plane displayed the highest peak intensity at a Bragg angle of 31°, indicating its significance in the crystalline structure of ZnO NPs. The Debye–Scherrer equation was used for computing the crystallite size (Eq. 2):

(2) D = K λ β cos θ

where D is the crystallite size of the NPs, K is the Scherrer constant (0.98), β is the full width at half-maximum of the diffraction peak, and λ is the wavelength of X-rays (1.54 Å). The average crystallite size was calculated to be 15.24 nm, suggesting that the synthesized ZnO NPs possess a relatively small size. Our finding aligns with the reports of Aklilu and Aderaw [48], which showed the crystallite size of ZnO NPs bio-synthesized using Catha edulis leaf extract was approximately 17 nm. The XRD analysis of bio-fabricated ZnO NPs prepared utilizing the leaf extract of Salvia officinalis, as detailed by Abomuti et al. [49], reveals important insights into the structural properties of the NPs. The analysis shows eminent peaks conforming to the crystallographic planes (100), (002), and (101), with 2θ values of 31.7°, 34.3°, and 36.2°, respectively. These peaks indicate the crystallinity of the synthesized ZnO NPs, which are characteristics of the hexagonal wurtzite structure, which is a common and stable form of ZnO. Comparable results were also observed for the biosynthesized ZnO NPs using Eucalyptus globulus Labill. leaf extract [50].

Further, the SEM and HR-TEM images show the surface morphology, shape, and size of the ZnO NPs (Figure 4). The SEM results reveal that ZnO NPs are hexagonal with a smooth surface. The EDX analysis confirms the occurrence of only zinc and oxygen, indicating high purity, and suggests that plant phytochemicals contribute to reducing and stabilizing the NPs. Similar results were described for ZnO NPs derived from plant extracts [51,52]. However, HR-TEM images provide the exact morphology and size of the synthesized ZnO NPs at different magnifications. The images obtained through TEM reveal the hexagonal shape and size within the range of 25–85 nm and an average diameter close to 50 nm. However, the Scherrer rings in the SAED pattern matched well with the XRD planes, which confirms the formation of crystalline ZnO NPs.

Figure 4 (a) SEM image, (b) EDX spectra, (c) HR-TEM image (inset: histogram depicting size distribution), and (d) SAED pattern of ZnO NPs.
Figure 4

(a) SEM image, (b) EDX spectra, (c) HR-TEM image (inset: histogram depicting size distribution), and (d) SAED pattern of ZnO NPs.

4.4 Molecular identification of the bacterial isolate

The isolated bacteria cultured on blood agar and MacConkey agar were confirmed to be E. coli through molecular identification. The submission of the nucleotide sequence was done in the NCBI’s GenBank with accession number (OR223289.1). The homology search using BLAST revealed 100% similarity with E. coli (AP027850.1).

4.5 Antibacterial activity of ZnO NPs

The T. spathacea synthesized ZnO NPs demonstrated antibacterial activity against E. coli isolated from a sputum sample. The clear inhibition zones obtained were found to be 9.6, 12.1, and 13.2 mm at concentrations of 0.25, 0.5, and 1 mg/disc, respectively. Streptomycin, which was used as a positive control, exhibited an inhibition zone of 17 mm, while sterile discs impregnated with distilled water showed no inhibition, as illustrated in Figure 5. Consistent with this study, various researchers have utilized the disc diffusion technique to evaluate the antibacterial properties of ZnO NPs against E. coli [53,54,55]. Chunchegowda et al. [56] demonstrated that ZnO NPs synthesized from Passiflora subpeltata effectively inhibited E. coli isolated from poultry feces with the inhibition zones measuring 17, 15, and 12 mm at doses of 1, 0.5, and 0.1 mg/disc, respectively. These results align closely with those of our current study.

Figure 5 (a) Antibacterial activity of ZnO NPs (the letters “N” and “P” indicate negative and positive control [streptomycin], while the numbers 1, 2, and 3 refer to the concentration of ZnO NPs at 1, 0.5, and 0.25 mg·mL−1, respectively), and (b) bar graph representing the zone of inhibition. The data are expressed as mean ± SE (n = 3) and analyzed by one-way ANOVA followed by Tukey’s test. *p < 0.05 indicates significant difference compared to the control.
Figure 5

(a) Antibacterial activity of ZnO NPs (the letters “N” and “P” indicate negative and positive control [streptomycin], while the numbers 1, 2, and 3 refer to the concentration of ZnO NPs at 1, 0.5, and 0.25 mg·mL−1, respectively), and (b) bar graph representing the zone of inhibition. The data are expressed as mean ± SE (n = 3) and analyzed by one-way ANOVA followed by Tukey’s test. *p < 0.05 indicates significant difference compared to the control.

4.6 ROS generation in bacterial cells

Metal oxide NPs exert an antibacterial mechanism through the generation of ROS, which ultimately leads to oxidative stress [57]. In our study, the DCFDA method was utilized to examine the ability of ZnO NPs to induce ROS-mediated bacterial inhibition. Non-fluorescent DCFHDA is oxidized in the presence of ROS and transforms into green-fluorescent dichlorofluorescein (DCF) [58]. As shown in Figure 6, E. coli cells treated with positive control (0.8 mM H2O2) showed highest ROS generation compared to non-treated cells, while the treatment with ZnO NPs (1, 0.5, and 0.25 mg·mL−1) showed dose-dependent increase in ROS generation. The obtained ROS results align well with the zone of inhibition studies, where the highest zone was observed on 1 mg·mL−1 ZnO NP treatment.

Figure 6 ROS generation induced by ZnO NPs in E. coli by the DCFDA method.
Figure 6

ROS generation induced by ZnO NPs in E. coli by the DCFDA method.

4.7 Trypan blue dye exclusion test

Trypan blue staining can be used to demarcate a clear difference between dead and live cells. This dye cannot permeate the intact cell membrane of a live bacterium but is easily taken up by the dead cells as their membrane is ruptured [59]. In the microscopic analysis, trypan blue staining results indicated that untreated cells remained viable and appeared colorless, whereas cells treated with ZnO NPs took up the dye and appeared blue (Figure 7), indicating the disruption of membrane integrity of bacterial cells. This outcome confirms the effective antibacterial activity of ZnO NPs. Consistent with our findings, another study used the trypan blue dye assay to evaluate the antibacterial ability of silver NPs against a various bacterial pathogens [60].

Figure 7 Trypan blue dye exclusion assay: (a) control and (b) ZnO NP-treated cells.
Figure 7

Trypan blue dye exclusion assay: (a) control and (b) ZnO NP-treated cells.

4.8 In vitro anticancer activity

4.8.1 MTT assay

The anticancer efficacy of T. spathacea-synthesized ZnO NPs was evaluated by MTT assay. It is a sensitive technique for determining cell viability, which is based on the conversion of yellow-colored soluble MTT dye into insoluble formazan, which is purple-colored by mitochondrial dehydrogenase enzymes of live cells; however, the dead cells do not express this activity [61]. In our study, we found that the tested concentration of ZnO NPs on HeLa cells showed dose-dependent cell viability ranging approximately from 88 to 24% at dosages of 10–320 µg·mL−1 (Figure 8a). The IC50 concentration of ZnO NPs was found to be 84.26 μg·mL−1. However, 10 μg of standard cisplatin (IC50 value) treatment revealed around 51% viability. Further, the anti-proliferative ability of ZnO NPs was confirmed by phase contrast microscopic images. The control group showed regular and healthy cell morphologies. However, the ZnO NP-treated cells showed decreased cell density with shrinkage and irregularly shaped and detached cells (Figure 8b). These results demonstrated the strong anticancer capability of the phytosynthesized ZnO NPs. The potent anticancer activity of these NPs is accredited to several mechanisms, including the profound generation of ROS, subsequently resulting in oxidative stress, mitochondrial dysfunction, damaged DNA, and induction of apoptosis, ultimately resulting in cell death [62]. Our findings are in line with the earlier reports, which also revealed the potent cytotoxic capacity of ZnO NPs synthesized using various plant extracts [63,64].

Figure 8 (a) Anti-cancer activity of ZnO NPs on HeLa cells by MTT assay (non-treated cells and cisplatin 10 μg·mL−1 served as negative and positive controls, respectively). The data are expressed as mean ± SE (n = 3) and analyzed by one-way ANOVA followed by Tukey’s test. *p < 0.05 indicates a significant difference compared to the control. (b) Morphological analysis using phase contrast microscopy.
Figure 8

(a) Anti-cancer activity of ZnO NPs on HeLa cells by MTT assay (non-treated cells and cisplatin 10 μg·mL−1 served as negative and positive controls, respectively). The data are expressed as mean ± SE (n = 3) and analyzed by one-way ANOVA followed by Tukey’s test. *p < 0.05 indicates a significant difference compared to the control. (b) Morphological analysis using phase contrast microscopy.

4.8.2 Cellular uptake of ZnO NPs

Confocal imaging was used for studying the cellular uptake of RITC-labeled ZnO NPs in HeLa cells. In our study, we used Hoechst 33342 for staining the nucleus. As shown in Figure 9, the nucleus appears blue, while red fluorescence indicates the internalization of RITC-labeled ZnO NPs within the HeLa cells. Further, the uptake is confirmed upon merging the bright field images with those obtained through blue (nucleus) and red channels (RITC-labeled ZnO NPs), which clearly show that the cell has sufficiently internalized ZnO NPs. Further, the mean fluorescence intensity of cytoplasmic staining (RITC-tagged ZnO NPs) was found to be 58.143. The process of cellular uptake involves two steps: first, the adherence of material (NPs) onto the cell and then interaction with lipids, proteins, and the other elements in the membrane of the cell. After this, uptake mechanisms are activated, which enhances the material’s absorption efficiency [65].

Figure 9 Cellular uptake of RITC-labeled ZnO NPs through confocal microscopy.
Figure 9

Cellular uptake of RITC-labeled ZnO NPs through confocal microscopy.

4.8.3 Assessment of MMP

JC-1 (fluorescent dye) was employed for the MMP measurements. JC-1 emits red fluorescence upon accumulating in the mitochondria of the healthy cells but produces green fluorescence upon leaching into the cytosol as a result of a reduction in MMP, leading to a negative internal potential [66]. Our results revealed that the control cells showed 93.2% and 6.41% red and green fluorescence, respectively, indicating intact MMP. Whereas the ZnO NP treatment (IC50 concentration) significantly reduced the red fluorescence to 79.9% along with an increase in green fluorescence up to 19.0%, confirming impairment in MMP (Figure 10). These results indicate that the treatment with ZnO NPs considerably disrupted the MMP, which can lead to the opening of the pores of the outer membrane and the release of apoptosis-modulating proteins, resulting in the activation of apoptotic cascade [67].

Figure 10 MMP of (a) control and (b) ZnO NP-treated cells.
Figure 10

MMP of (a) control and (b) ZnO NP-treated cells.

4.8.4 Apoptotic analysis

Apoptosis is a process of controlled cell death with apparent nuclear changes, resulting in the formation of apoptotic bodies [39]. Hoechst staining was utilized to identify the morphological aberrations in the nucleus, which is a clear indication of apoptosis [68]. After being exposed to ZnO NPs at their IC50 concentrations, the cells showed bright blue fluorescence with a condensed and fragmented nucleus, as indicated by the arrows (Figure 11). However, the control cells showed regular-shaped nuclei with a dull blue appearance indicative of viable and healthy cells. Further, this was validated using staining with annexin V-FITC and PI. Figure 12 clearly depicts the apoptosis-inducing capability of ZnO NPs with the early and late apoptotic percentages of 38.53% and 10.70%, respectively, while the viable cells were found to be 48.75% compared to 99.60% in control cells.

Figure 11 Morphological analysis of cell nucleus using Hoechst staining.
Figure 11

Morphological analysis of cell nucleus using Hoechst staining.

Figure 12 (a) Flow cytometric analysis of apoptosis using Annexin V-FITC and PI staining (LL, live cells, LR, early apoptotic cells, UR, late apoptotic cells, and UL, necrotic cells), and (b) bar chart representing the percentage of cells in different quadrants.
Figure 12

(a) Flow cytometric analysis of apoptosis using Annexin V-FITC and PI staining (LL, live cells, LR, early apoptotic cells, UR, late apoptotic cells, and UL, necrotic cells), and (b) bar chart representing the percentage of cells in different quadrants.

4.8.5 Cell cycle analysis

The cell cycle was meticulously regulated through a sophisticated network of regulatory molecules [69]. In our study, PI staining was utilized to observe the cell cycle distribution of HeLa cells. From Figure 13, it is apparent that the cell number in the sub-G0 phase augmented in the ZnO NP treatment group (5.02%) from 1.26% (control), indicating the increase in the apoptotic population. While the G0/G1 phase showed a significant reduction (53.7%) in treated cells compared to control (64.3%). However, the cells in the S and G2/M phases displayed moderate increases from 13.8% and 20.4% to 16.2% and 25.1%, respectively. Our results are in accordance with the previous studies of Sahu et al. [70], which revealed the total increase in the DNA content of the sub-G0/G1, S, and G2/M phases leads to apoptosis by disrupting the normal progression of the cell cycle.

Figure 13 Cell cycle analysis of (a) control and (b) ZnO NP-treated cells.
Figure 13

Cell cycle analysis of (a) control and (b) ZnO NP-treated cells.

5 Conclusion

In our study, an aqueous extract of T. spathacea leaf was utilized in the biosynthesis of ZnO NPs by solution combustion, which is an eco-friendly, reliable, and safe process. Several characterization techniques confirmed the ZnO NP formation with distinct optical properties, purity, and hexagonal wurtzite structure in nanosize. Moreover, significant antibacterial efficacy against the sputum-isolated E. coli was exhibited by these NPs. Further, the anticancer activity of these NPs was confirmed through MTT, MMP, apoptosis, and cell cycle analysis. Therefore, these NPs can be considered as a therapeutic strategy for cancer treatment and bacterial infections.

Acknowledgment

The authors extend their appreciation to the Deanship of Scientific Research at King Khalid University for funding this work through the Large Group Research Project under grant number RGP2/259/46. The authors wish to convey their gratitude to the Department of Microbiology and Biotechnology, Bangalore University, for providing the essential facilities required to complete this study.

  1. Funding information: The authors extend their appreciation to the Deanship of Scientific Research at King Khalid University for funding this work through the Large Group Research Project under grant number RGP2/259/46. The authors are grateful to the University Grants Commission (UGC) of India for funding under the UGC startup grant No. F.30-580/2021(BSR) dated 01.11.2021.

  2. Author contributions: Sumreen Sultana: writing – original draft, writing – review and editing, methodology, and investigation; Bagepalli Shivaram Ashwini: resources and writing – review and editing; Venkateshppa Bhavana: writing – original draft and investigation; Umme Hani, Nazima Haider, Mohammad Nasser Alomary, and Bangari Daruka Prasad: formal analysis; Tekupalli Ravikiran: formal analysis and visualization; Mohammad Azam Ansari and Thimappa Ramachandrappa Lakshmeesha: conceptualization, project administration, writing – review and editing, and supervision.

  3. Conflict of interest: Authors state no conflict of interest.

  4. Data availability statement: All data generated or analyzed during this study are included in this published article.

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Received: 2024-11-24
Published Online: 2025-06-10

© 2025 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  9. Preparation of Co/Cr-MOFs for efficient removal of fleroxacin and Rhodamine B
  10. Applying nanocarbon prepared from coal as an anode in lithium-ion batteries
  11. Improved electrochemical synthesis of Cu–Fe/brass foil alloy followed by combustion for high-efficiency photoelectrodes and hydrogen production in alkaline solutions
  12. Precipitation of terephthalic acid from post-consumer polyethylene terephthalate waste fractions
  13. Biosynthesized zinc oxide nanoparticles: Multifunctional potential applications in anticancer, antibacterial, and B. subtilis DNA gyrase docking
  14. Anticancer and antimicrobial effects of green-synthesized silver nanoparticles using Teucrium polium leaves extract
  15. Green synthesis of eco-friendly bioplastics from Chlorella and Lithothamnion algae for safe and sustainable solutions for food packaging
  16. Optimizing coal water slurry concentration via synergistic coal blending and particle size distribution
  17. Green synthesis of Ag@Cu and silver nanowire using Pterospermum heterophyllum extracts for surface-enhanced Raman scattering
  18. Green synthesis of copper oxide nanoparticles from Algerian propolis: Exploring biochemical, structural, antimicrobial, and anti-diabetic properties
  19. Simultaneous quantification of mefenamic acid and paracetamol in fixed-dose combination tablet dosage forms using the green HPTLC method
  20. Green synthesis of titanium dioxide nanoparticles using green tea (Camellia sinensis) extract: Characteristics and applications
  21. Pharmaceutical properties for green fabricated ZnO and Ag nanoparticle-mediated Borago officinalis: In silico predications study
  22. Synthesis and optimization of gemcitabine-loaded nanoparticles by using Box–Behnken design for treating prostate cancer: In vitro characterization and in vivo pharmacokinetic study
  23. A comparative analysis of single-step and multi-step methods for producing magnetic activated carbon from palm kernel shells: Adsorption of methyl orange dye
  24. Sustainable green synthesis of silver nanoparticles using walnut septum waste: Characterization and antibacterial properties
  25. Efficient electrocatalytic reduction of CO2 to CO over Ni/Y diatomic catalysts
  26. Greener and magnetic Fe3O4 nanoparticles as a recyclable catalyst for Knoevenagel condensation and degradation of industrial Congo red dye
  27. Recycling of HDPE-giant reed composites: Processability and performance
  28. Fabrication of antibacterial chitosan/PVA nanofibers co-loaded with curcumin and cefadroxil for wound healing
  29. Cost-effective one-pot fabrication of iron(iii) oxychloride–iron(iii) oxide nanomaterials for supercapacitor charge storage
  30. Novel trimetallic (TiO2–MgO–Au) nanoparticles: Biosynthesis, characterization, antimicrobial, and anticancer activities
  31. Green-synthesized chromium oxide nanoparticles using pomegranate husk extract: Multifunctional bioactivity in antioxidant potential, lipase and amylase inhibition, and cytotoxicity
  32. Therapeutic potential of sustainable zinc oxide nanoparticles biosynthesized using Tradescantia spathacea aqueous leaf extract
  33. Chitosan-coated superparamagnetic iron oxide nanoparticles synthesized using Carica papaya bark extract: Evaluation of antioxidant, antibacterial, and anticancer activity of HeLa cervical cancer cells
  34. Antioxidant potential of peptide fractions from tuna dark muscle protein isolate: A green enzymatic approach
  35. Clerodendron phlomoides leaf extract-mediated synthesis of selenium nanoparticles for multi-applications
  36. Optimization of cellulose yield from oil palm trunks with deep eutectic solvents using response surface methodology
  37. Nitrogen-doped carbon dots from Brahmi (Bacopa monnieri): Metal-free probe for efficient detection of metal pollutants and methylene blue dye degradation
  38. High energy density pseudocapacitor based on a nanoporous tungsten(VI) oxide iodide/poly(2-amino-1-mercaptobenzene) composite
  39. Green synthesized Ag–Cu nanocomposites as an improved strategy to fight multidrug-resistant bacteria by inhibition of biofilm formation: In vitro and in silico assessment study
  40. In vitro evaluation of antibacterial activity and associated cytotoxicity of biogenic silver nanoparticles using various extracts of Tabernaemontana ventricosa
  41. Fabrication of novel composite materials by impregnating ZnO particles into bacterial cellulose nanofibers for antimicrobial applications
  42. Solidification floating organic drop for dispersive liquid–liquid microextraction estimation of copper in different water samples
  43. Kinetics and synthesis of formation of phosphate composites from low-grade phosphorites in the presence of phosphate–siliceous shales and oil sludge
  44. Removal of minocycline and terramycin by graphene oxide and Cr/Mn base metal–organic framework composites
  45. Microfluidic preparation of ceramide E liposomes and properties
  46. Therapeutic potential of Anamirta cocculus (L.) Wight & Arn. leaf aqueous extract-mediated biogenic gold nanoparticles
  47. Review Article
  48. Sustainable innovations in garlic extraction: A comprehensive review and bibliometric analysis of green extraction methods
  49. Rapid Communication
  50. In situ supported rhodium catalyst on mesoporous silica for chemoselective hydrogenation of nitriles to primary amines
  51. Special Issue: Valorisation of Biowaste to Nanomaterials for Environmental Applications
  52. Valorization of coconut husk into biochar for lead (Pb2+) adsorption
  53. Corrigendum
  54. Corrigendum to “An updated review on carbon nanomaterials: Types, synthesis, functionalization and applications, degradation and toxicity”
https://doi.org/10.1515/gps-2024-0241
Keywords for this article
HeLa cells; Escherichia coli; green synthesis; mitochondrial membrane potential; apoptosis; cell cycle
Creative Commons
BY 4.0

Articles in the same Issue

  1. Research Articles
  2. Optimized green synthesis of silver nanoparticles from guarana seed skin extract with antibacterial potential
  3. Green adsorbents for water remediation: Removal of Cr(vi) and Ni(ii) using Prosopis glandulosa sawdust and biochar
  4. Green approach for the synthesis of zinc oxide nanoparticles from methanolic stem extract of Andrographis paniculata and evaluation of antidiabetic activity: In silico GSK-3β analysis
  5. Development of a green and rapid ethanol-based HPLC assay for aspirin tablets and feasibility evaluation of domestically produced bioethanol in Thailand as a sustainable mobile phase
  6. A facile biodegradation of polystyrene microplastic by Bacillus subtilis
  7. Enhanced synthesis of fly ash-derived hydrated sodium silicate adsorbents via low-temperature alkaline hydrothermal treatment for advanced environmental applications
  8. Impact of metal nanoparticles biosynthesized using camel milk on bacterial growth and copper removal from wastewater
  9. Preparation of Co/Cr-MOFs for efficient removal of fleroxacin and Rhodamine B
  10. Applying nanocarbon prepared from coal as an anode in lithium-ion batteries
  11. Improved electrochemical synthesis of Cu–Fe/brass foil alloy followed by combustion for high-efficiency photoelectrodes and hydrogen production in alkaline solutions
  12. Precipitation of terephthalic acid from post-consumer polyethylene terephthalate waste fractions
  13. Biosynthesized zinc oxide nanoparticles: Multifunctional potential applications in anticancer, antibacterial, and B. subtilis DNA gyrase docking
  14. Anticancer and antimicrobial effects of green-synthesized silver nanoparticles using Teucrium polium leaves extract
  15. Green synthesis of eco-friendly bioplastics from Chlorella and Lithothamnion algae for safe and sustainable solutions for food packaging
  16. Optimizing coal water slurry concentration via synergistic coal blending and particle size distribution
  17. Green synthesis of Ag@Cu and silver nanowire using Pterospermum heterophyllum extracts for surface-enhanced Raman scattering
  18. Green synthesis of copper oxide nanoparticles from Algerian propolis: Exploring biochemical, structural, antimicrobial, and anti-diabetic properties
  19. Simultaneous quantification of mefenamic acid and paracetamol in fixed-dose combination tablet dosage forms using the green HPTLC method
  20. Green synthesis of titanium dioxide nanoparticles using green tea (Camellia sinensis) extract: Characteristics and applications
  21. Pharmaceutical properties for green fabricated ZnO and Ag nanoparticle-mediated Borago officinalis: In silico predications study
  22. Synthesis and optimization of gemcitabine-loaded nanoparticles by using Box–Behnken design for treating prostate cancer: In vitro characterization and in vivo pharmacokinetic study
  23. A comparative analysis of single-step and multi-step methods for producing magnetic activated carbon from palm kernel shells: Adsorption of methyl orange dye
  24. Sustainable green synthesis of silver nanoparticles using walnut septum waste: Characterization and antibacterial properties
  25. Efficient electrocatalytic reduction of CO2 to CO over Ni/Y diatomic catalysts
  26. Greener and magnetic Fe3O4 nanoparticles as a recyclable catalyst for Knoevenagel condensation and degradation of industrial Congo red dye
  27. Recycling of HDPE-giant reed composites: Processability and performance
  28. Fabrication of antibacterial chitosan/PVA nanofibers co-loaded with curcumin and cefadroxil for wound healing
  29. Cost-effective one-pot fabrication of iron(iii) oxychloride–iron(iii) oxide nanomaterials for supercapacitor charge storage
  30. Novel trimetallic (TiO2–MgO–Au) nanoparticles: Biosynthesis, characterization, antimicrobial, and anticancer activities
  31. Green-synthesized chromium oxide nanoparticles using pomegranate husk extract: Multifunctional bioactivity in antioxidant potential, lipase and amylase inhibition, and cytotoxicity
  32. Therapeutic potential of sustainable zinc oxide nanoparticles biosynthesized using Tradescantia spathacea aqueous leaf extract
  33. Chitosan-coated superparamagnetic iron oxide nanoparticles synthesized using Carica papaya bark extract: Evaluation of antioxidant, antibacterial, and anticancer activity of HeLa cervical cancer cells
  34. Antioxidant potential of peptide fractions from tuna dark muscle protein isolate: A green enzymatic approach
  35. Clerodendron phlomoides leaf extract-mediated synthesis of selenium nanoparticles for multi-applications
  36. Optimization of cellulose yield from oil palm trunks with deep eutectic solvents using response surface methodology
  37. Nitrogen-doped carbon dots from Brahmi (Bacopa monnieri): Metal-free probe for efficient detection of metal pollutants and methylene blue dye degradation
  38. High energy density pseudocapacitor based on a nanoporous tungsten(VI) oxide iodide/poly(2-amino-1-mercaptobenzene) composite
  39. Green synthesized Ag–Cu nanocomposites as an improved strategy to fight multidrug-resistant bacteria by inhibition of biofilm formation: In vitro and in silico assessment study
  40. In vitro evaluation of antibacterial activity and associated cytotoxicity of biogenic silver nanoparticles using various extracts of Tabernaemontana ventricosa
  41. Fabrication of novel composite materials by impregnating ZnO particles into bacterial cellulose nanofibers for antimicrobial applications
  42. Solidification floating organic drop for dispersive liquid–liquid microextraction estimation of copper in different water samples
  43. Kinetics and synthesis of formation of phosphate composites from low-grade phosphorites in the presence of phosphate–siliceous shales and oil sludge
  44. Removal of minocycline and terramycin by graphene oxide and Cr/Mn base metal–organic framework composites
  45. Microfluidic preparation of ceramide E liposomes and properties
  46. Therapeutic potential of Anamirta cocculus (L.) Wight & Arn. leaf aqueous extract-mediated biogenic gold nanoparticles
  47. Review Article
  48. Sustainable innovations in garlic extraction: A comprehensive review and bibliometric analysis of green extraction methods
  49. Rapid Communication
  50. In situ supported rhodium catalyst on mesoporous silica for chemoselective hydrogenation of nitriles to primary amines
  51. Special Issue: Valorisation of Biowaste to Nanomaterials for Environmental Applications
  52. Valorization of coconut husk into biochar for lead (Pb2+) adsorption
  53. Corrigendum
  54. Corrigendum to “An updated review on carbon nanomaterials: Types, synthesis, functionalization and applications, degradation and toxicity”
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