Home Technology Eco-friendly green synthesis of silver nanoparticles with Syzygium aromaticum extract: characterization and evaluation against Schistosoma haematobium
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Eco-friendly green synthesis of silver nanoparticles with Syzygium aromaticum extract: characterization and evaluation against Schistosoma haematobium

  • Samah S. Eldera EMAIL logo , Esmat M. A. Hamzawy , Sarah Aldawsari , Reem Al-Wafi and Mohamed Abou El-Nour
Published/Copyright: November 20, 2025
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REVIEWS ON ADVANCED MATERIALS SCIENCE
From the journal REVIEWS ON ADVANCED MATERIALS SCIENCE Volume 64 Issue 1

Abstract

The use of nanotechnology in the treatment of many diseases is crucial to avoid the undesirable side effects of chemotherapy. Numerous attempts have been made to find alternative treatments using nanotechnology for these diseases. Schistosomiasis, caused by Schistosoma sp. parasites, is one of the most significant neglected tropical diseases in Africa, and the World Health Organization has approved a single chemotherapy treatment, Praziquantel (PZQ). Some reports have shown resistance of Schistosoma parasites to PZQ, and it has also been found to be ineffective against immature worms. Silver nanoparticles were synthesized using a green method to avoid the side effects of chemicals used in traditional methods, utilizing the plant Syzygium aromaticum, and their properties were consistent with previous studies. The green silver nanoparticles were evaluated in the laboratory at different concentrations: 3.125, 6.25, 12.5, 25, 50 and 100 μg/ml, as well as in mixtures with PZQ at concentrations of 12.5 + 0.4, 25 + 0.3, 50 + 0.2, and 75 + 0.1 μg/ml. At a concentration of 100 μg/ml, this resulted in complete worm death after 6 h, while concentrations of 50 and 25 μg/ml caused complete worm death after 12 h. A concentration of 12.5 μg/ml caused worm death after 24 h, while the effectiveness of PZQ was enhanced when mixed with silver nanoparticles, leading to faster killing. In in vivo study, hamsters were treated with the biosynthesized silver nanoparticles alone and in combination with PZQ, where both the size and number of granulomas, as well as the number of eggs in liver tissues, significantly decreased with treatment using silver nanoparticles.

Keywords: silver nanoparticles; green synthesis; Syzygium aromaticum ; Schistosoma haematobium

1 Introduction

Schistosoma haematobium is a neglected tropical parasite that causes schistosomiasis in humans. Despite the seriousness of this disease, which leads to great morbidity and causes the death of the patient, it has been neglected because it is directly linked to areas of unstable social and economic development, especially the loss of good sanitation. Schistosomiasis is spread in the Middle East, Southeast Asia and parts of South America and sub-Saharan Africa. Rating of those exposed to schistosomiasis are estimated at about 779 million people, and there are more than 250 million people who are actually infected with schistosomiasis, 201.5 million of whom live in Africa [1]. There are many species of schistosomes that infect humans, including S. haematobium, which causes urogenital disease and is endemic to the Middle East and Africa. Schistosomiasis leads to squamous cell carcinoma of the bladder, and for this reason, S. haematobium was classified as a group 1 carcinogen [2], Schistosoma mekongi, Schistosoma mansoni, Schistosoma malayensis and Schistosoma intercalatum these affects the large intestine; while Schistosoma japonicum, which affects the small intestine [3].

Because it is nowadays for a few ignored tropical diseases like leishmaniasis and trypanosomiasis, the therapy of schistosomiasis was nearly as troublesome and poisonous until the 1970s when Praziquantel was found [4], 5]. Praziquantel (PZQ) is the only chemotherapy drug recognized by the World Health Organization (WHO) for the treatment of schistosomiasis as it is safe and effective against all adult worms of all schistosomiasis infecting humans [4]. However, Praziquantel does not prevent reinfection and is much less effective in the pre-mature and juvenile stage, as it has been shown that some worms are resistant to this drug [5]. In addition, it causes immune responses and hypersensitivity [6]. Therefore, the goal of many researchers was to find an alternative medicine that would be safe and effective against schistosomiasis.

Nanomaterials have entered in recent years in many fields, including biomedicine, environment, energy sciences, chemical industries, space industries and cosmetics. The chemical synthesis of nanoparticles requires many physical and chemical pathways, including photochemical reduction, laser absorption, electrochemical reduction, and the Langmuir-Blodgett method, while this chemical synthesis forces researchers to use high radiation and stabilization agents, as well as toxic and reactive reducing agents [7], 8]. Because of the toxicity associated with chemical synthesis, it affects the environment, which negatively affects living organisms, which imposes various restrictions on the use of this method. Therefore, development in the production of nanoparticles by the green synthesis method, as it is environmentally friendly and cost-effective. This method promised a new era of safe nanomaterials technology production. One of the most important ways to produce green nanoparticles involves the use of living organisms (microorganisms and plants) [9]. Plants contain many chemical compounds, which makes them the best candidates to facilitate the green synthesis of nanoparticles because they can act as reducing, stabilizing and capping agents during the manufacturing process. While, the use of plants for the synthesis of nanoparticles is safer than chemical and bacterial methods because there is no threat of dangerous bacterial and chemical contamination and also by using less energy [10]. Silver nanoparticles have been used in many biological uses, including antibacterial, antifungal, anti-infective, antiviral, anti-inflammatory and wound-healing properties at low concentrations [11], 12]. Silver nanoparticles are an inorganic, non-toxic, antibacterial agent that can eliminate around 650 different types of pathogenic microorganisms [13], 14].

The pharmaceutical effects of silver nanoparticles on a variety of parasites were examined by green synthesis using a wide range of plant extracts, either in vitro or in vivo experiments; Leishmania sp. treated with Commiphora molmol (myrrh) [15], Plasmodium sp. treated with Azadirachta indica and Ocimum sanctum [16], Toxoplasma gondii treated with Ziziphus spinachristi and phoenix dactylifera [17] and Trichinella spiralis treated with C. molmol (myrrh) [18]. There have also been many attempts to use metals and metal oxides at the nanoscale to control and eliminate various microbes e.g. E.coli and Streptobacillus sp. treated with gold nanoparticles using aqueous fruit extract of Artocarpus heterophyllus [19], Aspergillus niger, Aspergillus flavus treated with gold nanoparticles using flower extract of Caesalpinia pulcherrima [20], Escherichia coli, Pseudomonas aeruginosa, Bacillus aureus and Staphylocccus aureus treated with silver nanoparticles using leaves extract of Justica wynaadensis [21]. Also, bimetallics (Fe–Pd and ZnO–CuO) were converted using the green method to nano size for use in various industries to improve their quality [22], 23].

The aim of the present research was to evaluate the harmful effect of green silver nanoparticles against S. haematobium (Egyptian strain) and how this is reflected in the survival rate.

2 Materials and methods

All used tools and glassware were washed with sterilized distilled water several times and dried in the oven to remove any contaminants before use. Silver nitrate (AgNO3, 99 %) and Whatman filter paper purchased from Sigma-Aldrich.

2.1 Aqueous extract preparation

Clove buds, Syzygium aromaticum were purchased from the local market, dried in the sun for 7 h to get rid of moisture in the buds, and then sieved to remove impurities. To obtain fine particles, the buds were ground and then sifted through a 10-mesh sieve. In 100 ml of hot sterilized distilled water, 25 g of sifted clove buds were dissolved. The solution was boiled for 30 min at 50 °C and left undisturbed at room temperature until it is cooled over a magnetic stirrer. The solution is filtered with Whatman 1 filter paper. Finally, the extracts were placed in porcelain dishes in temperature-controlled oven to remove the remaining water in the extracts to give a residue of 4.5 g. At 4 °C, the residues were stored for further use in a dark bottle.

2.1.1 Biosynthesis of silver nanoparticles

A 10 ml solution of silver nitrate (0.7 g of AgNO3 in a liter of double distilled water) was prepared. The reducing agent (aqueous extract of S. aromaticum) was added at a ratio of 3:1 with continuous stirring at a temperature of 90 °C using a thermomagnetic stirrer. As the reaction continued, the color of the solution changed from brown to golden yellow, indicating that the silver nitrate particles were reduced to the size of silver nanoparticles. The solution is cooled in a dark area of room temperature. The mixture is centrifuged at 10,000 rpm for 30 min. The silver particles were washed 4 times with double distilled water and dried by Lyophilizer.

2.2 Characterization of silver nanoparticles

For the analysis of interfaces to examine the surface adsorption functional groups on the synthesized silver nanoparticles by preparing the samples in the form of potassium bromide pellet (KBr), using the FT-IR spectrum (JASCO 6700 spectrometer) at the Egyptian Petroleum Research Institute (EPRI), Cairo, Egypt, the samples were scanned against a blank KBr pellet background at wave number range 4,000–400 cm−1. The size distribution and zeta potential measurements were determined using Malvern zetasizer Nano ZS, by injecting 2 ml of the solution into the cell, using a laser, at an angle of 173° to collect the feedback optics, where the applied potential difference to push the electrodes was 150 V of capillary electrophoresis. To examine silver nanoparticles through TEM (JEOL, model 1200EX, Tokyo, Japan), a copper grid (300 mesh) is used, a drop of suspension is placed on it, covered with a carbon film, and left to dry at room temperature. Silver nanoparticles were subjected to X-ray diffraction by Ultima IV (Rigaku, Japan), using Cu-Kα radiation with a wavelength (λ) of 1.5406 Å. X-ray scanning was carried out in 2/θ continuous mode at 2° per minute.

2.3 Toxicological study

Cytotoxicity was determined using the MTT colorimetric technique by adopting procedures as previously described (Ebrahimzadeh et al.) [24]. In a 96-well tissue culture plate, 1 × 105 cells/ml of Vero cell were inoculated for 24 h at 37 °C. After cell growth, the growth medium was discarded and the cells were washed twice with wash medium. Silver nanoparticles (31.25, 62.5, 125, 250, 500, 1,000 μl/ml) were diluted in Minimal Essential Medium (MEM, Gibco, USA) medium with 2 % serum. All wells were treated with different concentration of silver nanoparticles, leaving 3 wells as a negative control. The plate was incubated at 37 °C. Add 20 ml of MTT solution to each well and mix well. Incubate for 4 h at 37 °C and 5 % CO2 to allow MTT metabolism. The culture is completely discarded. Add 200 ml of formazan and leave on a shaking table for 5 min mix the formazan well with the solvent. Using a spectrophotometer (Biotek Instruments, USA) the optical density of each well was measured individually at 590 nm. Cell viability % was estimated using the following equation:

Cell viability  % = Optical   density   OD sample OD   control × 100

3 Experimental design

3.1 Hamster and infection with Schistosoma

In this experiment, 60 males of golden hamster (Mesocricetus auratus) were used, some of which were used to obtain adult worms of S. haematobium for in vitro experiment, and others for in vivo experiment. The age of the hamster ranges from 7 to 9 weeks, and their weight ranges from 110 to 135 g. Hamsters were purchased from the Schistosome Biological Supply Program (SBSP), TBRI, Giza, Egypt. Hamsters were placed in polycarbonate boxes with wooden sawdust and covered with steel wire. In each box, there were 6 hamsters. The temperature was 22 ± 3 °C and the humidity was 50 ± 15. Feeding was available at all times and treatments and dissection were performed according to the ethical guidelines of the Theodor Bilharz Research Institute (TBRI). Hamsters were infected according to Smithers and Terry [25] by shaving part of the abdominal hair and placing them on a cotton wool in contact with water containing 250 live cercariae for 30 min until infection occurred. 90 days after infection, according to Smithers and Terry [25] five hamsters were sacrificed by cervical decapitation, then perfusion of the porto-mesenteric and hepatic vessels was achieved to count all recovered worms.

3.2 In vitro study

The culture medium was prepared consisting of Roswell Park Memorial Institute (RPMI) 1,640 culture medium (Biowhittaker, Lonza, B-4800 verviers, Belgium) with an antibiotic to inhibit the growth of bacteria (streptomycin 100 μg/ml, gentamycin 160 μg/ml and penicillin 100 U/ml), 20 % fetal calf serum (Gibco, USA), 2 g/l glucose, 20 g/l Na HCO3 of medium and 0.39 g/l glutamate. In 24-well culture plates (TPP, ST. Louis, Mo.), four groups were exposed to different concentrations, and in each well, seven adult couple worms of S. haematobium were placed. The first group of adult worms of S. haematobium were exposed to different concentrations of silver nanoparticles (AgNPs) (3.125, 6.25, 12.5, 25, 50, and 100 μg/ml). The second group of adult worms were exposed to different concentrations of silver nanoparticles (AgNPs) + PZQ (12.5 + 0.4, 25 + 0.3, 50 + 0.2, 75 + 0.1 μg/ml). The fourth group of adult worms were exposed to a concentration of 0.5 μg/ml of PZQ as a positive control. The fifth group of worms were exposed to dimethyl sulfoxide (DMSO) as a negative control. Each well contained 2 mL of medium. The plates were incubated for 48 h in a humidified atmosphere of 5 % carbon dioxide, and at a temperature of 37 °C (Thermo Fisher Scientific, Marietta, OH, USA). Using an Olympus optical inverted microscope, the worms were observed at intervals of 2, 4, 6, 12, 24 and 48 h. The experiment was repeated three times. All steps of the experiment were done in a sterile laminar flow chamber. the treated worms were examined for their pairing (mating), and locomotion (the worms’ motor activity changes). Worms are monitored for 2 min, and if no movement appears, they become shrunken and change their color to black, they are considered dead.

3.2.1 Electron microscope

To examine the changes that occurred on the integument of adult male and female worms of S. haematobium after incubating for 48 h with different concentrations of the tested materials, they were examined by scanning electron microscope (Inspect S; FEI, Holland) at the Regional Center of Mycology and Biotechnology (RCMB). For preparation samples were preserved in 3 % glutaraldehyde in 0.2 % sodium cacodylate solution for 120 min. They were washed for 120 min with a solution of 0.2 % cacodylate and 0.4 % sucrose in equal volume. The samples were transferred for 60 min in a solution of 2 % osmic acid and 0.2 % cacodylate in equal volume, then washed with distilled water. The samples are dried with an ascending series of ethyl alcohol and this step is repeated three times. Schistosomes worms are placed on metal thump, coated with gold for examination.

3.3 In vivo study

3.3.1 Golden hamsters

Hamsters previously infected with S. haematobium cercariae began to excrete Schistosoma eggs after the worms reached adulthood. The hamsters were categorized into four groups (5 hamsters per group), while the fifth group consisted of hamsters uninfected with S. haematobium, serving as the healthy control group. The initial cohort, hamsters infected with S. haematobium, received treatment with distilled water (negative control group). The second group, consisting of hamsters, received a treatment of 600 mg/kg of body weight of Biltersed suspension (Alexandria Company for Pharmaceuticals and Chemical, Alexandria, Egypt) as the positive control group. The third group, the hamsters, received a dosage of 50 mg/kg of body weight of silver nanoparticle suspension (Treatment Group 1). The fourth group of hamsters received a treatment consisting of 50 mg/kg body weight of silver nanoparticles suspension combined with 300 mg/kg body weight of biltersed suspension (Treatment group 2). All doses were administered orally. Three replicates were conducted in the experiment from the third and fourth groups.

3.3.2 Histopathological assessment

Subsequent to the sacrifice of the hamsters, the liver was excised and preserved in a 10 % saline solution, after which the samples were maintained in 10 % formalin. The chosen specimens are cleansed to eliminate surplus formalin, and the tissues are desiccated by subjecting them to an ascending series of alcohol at hourly intervals. Xylene is utilized to eliminate excess alcohol, followed by waxing with paraffin, and subsequently allowed to dry for sectioning. The tissue is sectioned to a thickness of 5 μm using a microtome, and the wax is eliminated with xylene. Tissues undergo staining with haematoxylin and eosin (H&E) subsequent to rehydration. The tissues were analyzed using an inverted light microscope to identify granuloma following the application of DPX.

3.4 Statistical analysis

The statistical software SPSS Version 22 was utilized for data coding and entry. Continuous variables underwent the Shapiro-Wilk and Kolmogorov-Smirnov tests for normality, and the data were evaluated to determine compliance with the assumptions of parametric tests. The arcsine square root transformation was employed to standardize probability and percentile data for normality assessment. The data’s mean and standard deviation were presented. ANOVA analyses were conducted concerning the recorded mortality among the experimental groups (Control, Lemongrass LC15, Lemongrass LC50, Citral LC15, and Citral LC50). The analysis was conducted with a minimum of three replicates for each group. The post-hoc analysis was conducted utilizing a Tukey pairwise comparison. Data were presented using R Studio version 2022.02.4 when possible.

4 Results

4.1 Green synthesis of silver nanoparticles (AgNPs)

As a reducing and capping agent, S. aromaticum extract was used to synthesized AgNPs. When added to plant extract, the colorless silver nitrate solution changes to a dark brown color. During the reaction process, the solution’s color changes from light yellow to brownish yellow, indicating the reduction of silver ions by the plant extract to zero valence silver particles and indicating the formation of silver nanoparticles.

4.2 Characterization of green synthesized silver nanoparticles

4.2.1 FT-IR analysis

The reduction of metal Ag+ to Ag0 nanoparticles was facilitated by the functional groups present in Eucalyptus citratus extract, which play roles as capping, stabilizing, and reducing agents. FT-IR analysis was conducted to examine potential interactions between these functional groups and the green-synthesized AgNPs in the 400 cm−1 to 4,000 cm−1, as shown in Figure 1.

Figure 1: FTIR spectra of green synthesized silver nanoparticles by Syzygium aromaticum.
Figure 1:

FTIR spectra of green synthesized silver nanoparticles by Syzygium aromaticum.

The main and strongest vibration modes of the greenly synthesized AgNPs were detected in the FTIR data at approximately 3,348.26 cm−1, 2,986.39 cm−1, 2,164.49 cm−1, 1,598.65 cm−1, 1,374.38 cm−1, 1,045.46 cm−1, 822.88 cm−1, and 594.78 cm−1. The broad band at 3,348.26 cm−1 is caused by the amino groups (NH2) present in the S. aromaticum extract vibrating in the N–H stretching, which corresponds to the –OH stretching of phenolic compounds. The peak at 2,986.39 cm−1 is associated with C–H stretching modes, whereas the peak at 1,598.65 cm−1 is associated with N–H binding primary amides and is associated with C=O stretching, indicating the presence of compounds containing carboxylates, aldehydes, esters, ketones derived from tannins and flavonoids. The N–H stretching vibration present in the amide linkage is demonstrated by an intense peak at 1,374.38 cm−1, which is typical for silver metal. The C–N stretching vibrations of the aliphatic amines could be attributed to the extreme range at 1,045.46 cm−1. As shown in Figure 1, the peak at 822.88 cm−1 and 594.78 cm−1 represents strong bands of alkenes and aromatic ring, respectively.

4.2.2 Transmission electron microscope (TEM)

The size of the biosynthesis silver nanoparticles in the present investigation was tested using TEM (Figure 2). The silver nanoparticles were homogeneously distributed and semi-spherical with a size of less than 70 nm. This result is confirmed by the results obtained from the dynamic light scattering (DLS).

Figure 2: TEM of green synthesized silver nanoparticles by Syzygium aromaticum.
Figure 2:

TEM of green synthesized silver nanoparticles by Syzygium aromaticum.

4.2.3 XRD pattern analysis

XRD pattern analysis for the prepared AgNPs was shown in Figure 3. The reflection planes recorded from 20° to 60°. Two strong Bragg reflections for Ag at 38.25° and 44.45° correspond to the (111) and (200) planes, respectively, that can be indexed based on the characteristics of the face-centered cubic (FCC) crystal structure of silver. The interplanar spacing (calculated in Table 1) values are 2.353 Å, 2.038 Å for the (111) and (200) planes, respectively, the spectra closely align with the standard values recorded of Ag nanocrystals (JCPDS card no: 65–2,871). Moreover, appearance of minor peaks (marked stars) in the graph at 23.65°, 27.45°, 29.56°, 31.88°, 34.00°, related to the bio-organic phase that is present on the surface of the silver nanoparticles crystallizes, giving rise to some unassigned peaks.

Figure 3: X-ray diffraction of biosynthesized AgNPs by Syzygium aromaticum.
Figure 3:

X-ray diffraction of biosynthesized AgNPs by Syzygium aromaticum.

Table 1:

Calculated XRD data for biosynthesis silver nanoparticles by Syzygium aromaticum.

Peak Number Pos. [o2θ] d-spacing [Å] Height [cts] FWHM [°2θ] Rel. Int. [%]
1 23.6506 3.76198 26.30 0.1771 10.63
2 27.4525 3.24901 18.23 0.1771 7.37
3 29.5675 3.02125 20.84 0.2952 8.42
4 31.8852 2.80673 23.65 0.1771 9.56
5 34.0055 2.63643 12.62 0.4723 5.10
6 38.2507 2.35303 247.41 0.2657 100.00
7 44.4504 2.03818 68.67 0.2952 27.76

The Debye-Scherrer’s equation was used to determine the average particle crystal size diameter D.

D = K λ β cos θ

θ is the diffraction angle, λ is X-ray wavelength (0.1541 nm), β is the full width at half maximum (FWHM) and K is the Scherrer constant or shape factor (0.9). By calculating the average crystalline size of synthesized silver nanoparticles according to the Debye-Scherrer’s equation, it was found 39 nm, as shown in Table 1.

4.2.4 Zeta potential

The results showed that the zeta potential value of silver nanoparticles synthesized by clove extract was −31.0 mV (Figure 4). The synthesized AgNPs are kept stable by the negative charges, which prevent nanoparticle agglomeration. Zeta potential value range of 0–5 mV the nanoparticles are unstable due to their tendency to aggregate, and in range of 5–20 mV the nanoparticles are stable in a minimum, and in range of 20–40 mV the particles are stable, and in the range above 40 they are very stable. This demonstrates the stability of the silver nanoparticles acquired from S. aromaticum extract.

Figure 4: Zeta potential of green synthesized silver nanoparticles by Syzygium aromaticum.
Figure 4:

Zeta potential of green synthesized silver nanoparticles by Syzygium aromaticum.

4.2.5 Dynamic light scattering (DLS)

Using a dynamic light scattering particle size analyzer, the size of the synthesized silver nanoparticles was determined. Dynamic light scattering analysis of silver nanoparticles indicated their successful conversion to nano-size. The average hydrodynamic size of the silver particles, which were reduced and capping by S. aromaticum extract, was about 175.6 nm with a polydispersity index (PDI) of 0.277, as shown in DLS diagram of the synthesized Ag nanoparticles (Figure 5).

Figure 5: DSL of green synthesized silver nanoparticles by Syzygium aromaticum.
Figure 5:

DSL of green synthesized silver nanoparticles by Syzygium aromaticum.

4.3 In vitro study

The efficacy of the treatment in vitro for green-synthesized silver nanoparticles on adult S. haematobium worms at concentrations of 3.125, 6.25, 12.5, 25, 50, and 100 μg/ml was studied. The silver nanoparticles exhibited various clear effects that were examined to determine their potential impacts. Additionally, the effect of silver nanoparticles mixed with PZQ at concentrations of 12.5 + 0.4, 25 + 0.3, 50 + 0.2, and 75 + 0.1 μg/ml was also studied. The silver nanoparticles had a dose-dependent effect on each parameter evaluated. Movement, a significant decrease in the movement of adult worms was observed in most concentrations. The percentage of adult worms whose movement decreased was directly related to the incubation period and concentration. The movement activity of all adult worms was monitored after time periods of 2, 4, 6, 16, 24, and 48 h. The concentrations of 3.125 and 6.25 μg/ml of silver nanoparticles began to reduce the movement of the worms after 12 h without complete noticeable loss of movement. In the negative control groups, it was observed that the movement of the worms remained unchanged for 24 h, however, movement decreased over the 48-h period, but there was no complete loss of movement. Moreover, the effect of 0.5 μg/ml of PZQ (positive control group) led to a decrease in the movement activity of adult worms during the first 2 h of incubation, and complete loss of activity occurred in all worms within the four-hour period. While the previous concentrations did not significantly affect the stability of the mating of adult worms, as they separated after 40 h. Praziquantel (0.5 μg/ml) as a positive control group caused the separation of a pair of adult worms after the first 2 h of incubation. On the other hand, separation of the pairs was observed after, approximately, 40 h of incubation in the negative control groups. Additionally, it was confirmed that the concentrations that were not 99 % lethal to the parasites were effective in inhibiting the couples in all samples. In contrast, concentrations of 12.5 and 25 μg/ml of silver nanoparticles caused the separation of 30 % and 45 % of adult worms, respectively, and led to a decrease in worm movement after 12 and 6 h, respectively. These concentrations also killed 100 % of the adult worms after 24 and 12 h, respectively, with a statistically significant difference (p < 0.001). When the same concentrations were combined with PZQ (12.5 + 0.4 and 25 + 0.3 μg/ml), they caused the separation of 70 % and 100 % of the adult worms in the first 2 h, respectively, and led to a reduction in worm movement after 6 and 4 h, respectively. They also killed 100 % of the adult worms after 12 and 6 h, respectively, with a statistically significant difference (p < 0.001). At concentrations of 50 and 100 μg/ml of silver nanoparticles, a 100 % separation of adult worms occurred within the first 2 h, with a significant decrease in movement after 4 h of exposure. Nanoparticles of silver killed 100 % of the adult worms within 12 and 6 h, respectively, with a statistically significant difference (p < 0.001). At concentrations of 50 + 0.2 and 75 + 0.1 μg/ml of silver nanoparticles mixed with PZQ, 100 % separation of adult worms was achieved during the first 2 h, with a significant decrease in movement after the 2 h. The mortality rate reached 100 % within 4 h, with the mortality rate reached 100 % within 4 h, with a statistically significant difference (Tables 2 and 3) and (Figure 6a and b).

Table 2:

Statistical analysis of the effect of AgNPs synthesized by Syzygium aromaticum and AgNPs plus PZQ with different concentration and different times on male adult worms of S. haematobium.

Tested materials Conc. (µg/ml) Number of dead male adult worms of S. haematobium after incubation for 48 h
2 h 4 h 6 h 12 h 24 h 48 h
Green silver NPs 3.125 0 0 0 1 ± 0.81 2.66 ± 0.74 4 ± 0.81
6.25 0 0.66 ± 0.47 1 ± 0.81 1.66 ± 0.47 4 ± 0 5 ± 0
12.5 0.66 ± 0.47 2 ± 0 3.33 ± 0.94 4.66 ± 1.24 7 ± 0 7 ± 0
25 1 ± 0.81 3 ± 0.81 5.33 ± 0.94 7 ± 0 7 ± 0 7 ± 0
50 1 ± 0 3.66 ± 0.47 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0
100 1.33 ± 0.47 4 ± 0 7 ± 0 7 ± 0 7 ± 0 7 ± 0
Green silver NPs

+

PZQ		 12.5 + 4 2 ± 0.81 3.66 ± 0.47 5.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0
25 + 3 2.66 ± 0.47 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0
50 + 2 3.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0 7 ± 0
75 + 1 3 ± 0.81 7 ± 0 7 ± 0 7 ± 0 7 ± 0 7 ± 0
PZQ 0.5 μg/ml 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0 7 ± 0
Table 3:

Statistical analysis of the effect of AgNPs synthesized by Syzygium aromaticum and AgNPs plus PZQ with different concentration and different times on female adult worms of S. haematobium.

Tested materials Conc. (µg/ml) Number of dead female adult worms of S. haematobium after incubation for 48 h
2 h 4 h 6 h 12 h 24 h 48 h
Green silver NPs 3.125 0 0 0 1 ± 0 2.66 ± 0.94 3.66 ± 0.47
6.25 0 0.33 ± 0.47 1.33 ± 0.47 1.66 ± 0.47 3.33 ± 0.94 5.66 ± 0.47
12.5 0.66 ± 0.47 2.33 ± 0.94 4 ± 0.81 4.66 ± 1.24 7 ± 0 7 ± 0
25 0.66 ± 0.47 3.33 ± 0.47 5.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0
50 1 ± 0.81 3.66 ± 0.47 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0
100 1 ± 0 3.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0
Green silver NPs

+

PZQ		 12.5 + 4 2 ± 0.81 3.66 ± 0.47 6 ± 0.81 7 ± 0 7 ± 0 7 ± 0
25 + 3 3.33 ± 0.47 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0
50 + 2 4.33 ± 1.24 7 ± 0 7 ± 0 7 ± 0 7 ± 0 7 ± 0
75 + 1 3.33 ± 0.47 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0
PZQ 0.5 μg/ml 6.66 ± 0.47 7 ± 0 7 ± 0 7 ± 0 7 ± 0 7 ± 0

4.4 Tegumental topographic examination of S. haematobium by Scanning Electron Microscope (SEM)

Examining normal adult S. haematobium worms taken from untreated golden hamsters revealed that, the oral sucker of schistosome worms has an oval shape and is covered in acute spines of various sizes. The ventral sucker is the circular structure with spines behind the oral sucker. The abdominal surface thickens and ventrally folds behind the ventral sucker to create the gynaecophoric channel. Rows of tiny spines are present on the ventral side of Schistosoma. Male Schistosoma have a large number of tubercles on their dorsal surface, but there are no spines on the surface of the body in between them (Figure 7A and B).

Figure 6: Statistical analysis of the effect of AgNPs synthesized by Syzygium aromaticum and AgNPs plus PZQ with different concentration and different times on the adult worms of S. haematobium, (a) Male and (b) Female.
Figure 6:

Statistical analysis of the effect of AgNPs synthesized by Syzygium aromaticum and AgNPs plus PZQ with different concentration and different times on the adult worms of S. haematobium, (a) Male and (b) Female.

Figure 7: SEM photomicrograph of S. haematobium: (A and B) untreated worms (A) normal body architecture with numerous tubercles supported with acute spine, (B) normal oral sucker (oval) and rounded ventral sucker. (C and D) treated group with PZQ (C) severe damage of body, (D) tubercles deformation and spine decay (E, F, G, H) treated group with biosynthesized Ag nanoparticles (E) complete destruction of oral and ventral suckers, (F) peeling and destruction of the tegument (G) abnormalities appeared in the tubercles and destruction of the spines, (H) wrinkles and bubbles appeared on damage tegument, (I, J, K, L) treated group with AgNPs plus PZQ (I) severe damage of oral and ventral sucker appear pits (J) appearance pits on the surface with microtubules (K and L) completely destroyed of tegument (peeling of tubercles, tegument sloughing, devastation spines) (L) numerous bubbles appear.
Figure 7:

SEM photomicrograph of S. haematobium: (A and B) untreated worms (A) normal body architecture with numerous tubercles supported with acute spine, (B) normal oral sucker (oval) and rounded ventral sucker. (C and D) treated group with PZQ (C) severe damage of body, (D) tubercles deformation and spine decay (E, F, G, H) treated group with biosynthesized Ag nanoparticles (E) complete destruction of oral and ventral suckers, (F) peeling and destruction of the tegument (G) abnormalities appeared in the tubercles and destruction of the spines, (H) wrinkles and bubbles appeared on damage tegument, (I, J, K, L) treated group with AgNPs plus PZQ (I) severe damage of oral and ventral sucker appear pits (J) appearance pits on the surface with microtubules (K and L) completely destroyed of tegument (peeling of tubercles, tegument sloughing, devastation spines) (L) numerous bubbles appear.

The worms treated with PZQ showed destruction of the oral and ventral suckers. There was severe shrinkage in both the female and male tegument, with extensive peeling of the surface, which led to exposure of the underlying muscle layer. Additionally, the sensory papillae lost their dome-like shape and the terminal sensory spines became subsided. Edema was observed in the tegument around the oral sucker with ulcerations in the form of holes. Complete loss of the papillary spines was observed (Figure 7C and D).

The adult worms treated with green silver nanoparticles alone exhibited similar effects to those treated with PZQ. Some areas of the tegument appeared as irregular masses sub-integument inclusion, with general deformities in the papillae, partial and complete loss of papillary spines, and degeneration of the papillae. Additionally, many of the papillae showed collapse with extensive membranous folds between them and swollen areas in the male worms. Severe shrinkage was observed on most of the tegumental surface of the adult worms, accompanied by massive deformities in the oral and ventral suckers. Various-sized spherical blebs appeared in many tegumental areas. Furthermore, the gynaechophoric canal appeared deformities changes, such as pits that appear on the inner surface, were induced on the tegumental surface by the activity of silver nanoparticles (Figure 7E–H).

In the group treated with a mixture of silver nanoparticles and PZQ, there were severe deformities in the oral and ventral suckers with high contractions of the tegument. Numerous bubbles were observed in some surface areas, and the papillae lost their typical shape, with sensory spines either disappearing or shortening (Figure 7I–L).

4.5 Histopathological changes of liver architecture

The healthy group (control group) liver was histologically examined using light microscopy. Hepatocytes extend from the central vein of the hepatic lobes to the hepatic lobules’ periphery, where the portal tracts are visible. Additionally, there were no indications of inflammatory infiltrates (Figure 8A and B).

Figure 8: Light microscopy of liver sections from golden hamsters. (A and B) Show a normal (uninfected) liver section. (C and D) Depict a liver section infected with S. haematobium (untreated), characterized by numerous fibrocellular granulomas. (E–H) Illustrate an infected liver section treated with green AgNPs, revealing fibrocellular granulomas with miracidia inside the eggs scattered throughout the hepatic parenchyma. There is a marked reduction in both the number and size of granulomas, improved liver structure, egg destruction, and the presence of empty granulomas. The granulomas appear irregular in shape and contain less fibrous material. (I–L) Present an infected liver section treated with green AgNPs combined with PZQ, showing a significant decrease in granuloma number and size, enhanced liver architecture, egg destruction, and empty granulomas. The granulomas maintain an irregular shape with diminished fibrous content.
Figure 8:

Light microscopy of liver sections from golden hamsters. (A and B) Show a normal (uninfected) liver section. (C and D) Depict a liver section infected with S. haematobium (untreated), characterized by numerous fibrocellular granulomas. (E–H) Illustrate an infected liver section treated with green AgNPs, revealing fibrocellular granulomas with miracidia inside the eggs scattered throughout the hepatic parenchyma. There is a marked reduction in both the number and size of granulomas, improved liver structure, egg destruction, and the presence of empty granulomas. The granulomas appear irregular in shape and contain less fibrous material. (I–L) Present an infected liver section treated with green AgNPs combined with PZQ, showing a significant decrease in granuloma number and size, enhanced liver architecture, egg destruction, and empty granulomas. The granulomas maintain an irregular shape with diminished fibrous content.

Histopathological characteristics of schistosomiasis were apparent in liver sections from the infected, untreated group. The features comprised numerous dispersed fibro-cellular granulomas, inflammatory infiltrates encircling the portal spaces, plasma cells, densely clustered concentric aggregations of lymphocytes, hepatic stellate cells, eosinophils, and macrophages encircling entrapped eggs (Figure 8C and D).

The liver tissue cells of the infected group treated with Ag nanoparticles showed moderate infiltration by chronic inflammatory cells, but there were no fibrosis areas or Schistosoma eggs present. This group exhibited reduced density of connective tissue and a lower quantity of centrally located eggs. In comparison to the untreated positive control group, which exhibited a percentage reduction in granule count (43.1 %) and size (42.8 %), the absence of Schistosoma eggs and fibrosis, along with a notable decrease in chronic inflammatory liver cell infiltration, were observed (Table 4; Figure 8E–H). The cohort administered a combination of silver nanoparticles and PZQ exhibited the most significant percentage reduction in both the quantity (73.2 %) and dimensions of granules (63.3 %) as illustrated in Table 4 and Figure 8I–L.

Table 4:

The mean size and number of granuloma of hamster liver in infected control group comparison to treated group.

Groups of animals Diameter of granuloma (mean ± SE) Granuloma size reduction % Count of granuloma (mean ± SE) Count reduction %
Untreated control 356.71 ± 30.23 – 16.6 ± 5.43 –
Positive control 185.67 ± 31.25a 47.9 8.56 ± 4.43 48.4
Green AgNPs 203.73 ± 32.54a 42.9 9.87 ± 3.56 40.5
Green AgNPs + PZQ 131.74 ± 30.34a 63.1 5.63 ± 3.22 66.1

  1. aStatistically significant difference from infected control at P-value < 0.001.

5 Discussion

The main properties that determine the biological activity of nanoparticles are the shape, type, aggregation and stability of nanoparticles, including the toxic effect on biological systems.

The first step to form nanoparticles by green synthesis is to mix the biological extract with the metal salt solution. Chemicals in plant extracts are reducing agents in green synthesis. Biomolecules such as terpenoids and polyphenols can reduce metals by transferring electrons into metallic ions [26], 27]. As a result, reduced ions begin to form nucleus, which is a re-formation of ordered arrangements resembling a crystalline structure. The sizes of the nanoparticles increase with the stability of the reduced ions on the surface of the nucleus [28]. The growth of the particles is stopped and their size is stabilized by the biomolecules, which are attached to the surface of the particles and cover them, which prevents agglomeration among themselves [29]. For instance, the S. aromaticum extract’s saponins interact with silver ions (Ag+) through its hydroxyl groups to form silver nanoparticles, which give the newly formed silver nanoparticles stability. The carbonyl functional groups on the flavonoids in the plant extract interact with the metallic ions, converting the enols in the flavonoids into keto by reactive hydrogen liberation, which results in the formation of silver nanoparticles (Ag0) [10].

FTIR has become an important tool in analyzing silver binding molecules and understanding the participation of functional groups in plant extracts in the relationship between metal particles and biomolecules. FTIR can give accuracy, repeatability and good transmission ratio, and determine whether functional groups are related to the conversion of metal particles to metal nanoscale.

On the reduced nanoparticles, FTIR spectroscopy measurements were made to find potential biomolecules in the S. aromaticum extract that were in charge of reducing, capping, and stabilizing silver metal particles. The presence of peaks in the FTIR spectrum suggests that the AgNPs carry mainly coated functional group fragments from the plant extract. The reduction of metal salts in the plant extract is brought on by the high concentrations of flavonoids (gallic acid and ascorbic acid) and antioxidants [10]. Tannins and flavonoids serve as stabilizing and coating agents during green synthesis, and terpenoids are effective at converting aldehyde groups into carboxylic acids in silver metal ions. Additionally, polyphenols are potential reducing agents in the synthesis of AgNPs, and amide groups are responsible for the presence of enzymes that reduce the synthesis of silver metal ions and stabilize them. Additionally, the amino acid residue and protein carbonyl group have a stronger affinity for the metal and may prevent agglomeration [30].

The XRD pattern of the biosynthesized particles from the S. aromaticum extract was examined to confirm the crystalline nature of the silver nanoparticles. The values of the reported peaks agree with several previous results [7], 8]. According to Choudhary et al. [30] XRD showed the angle (2θ) peak at 38.2° which indicated the pure silver nanoparticles. Some non-specific peaks were observed because the clove buds extract contains some proteins and biological organic compounds that crystallize on the surface of the silver particles. Similar results synthesized of the silver nanoparticles using Coleus aromaticus extract, Mangifera indica extract and Geranium mushroom extract were reported [7], [8], [9].

The zeta potential is the difference in potential between the mobile dispersion medium and the stationary layer of the dispersion medium that is attached to the dispersed particle. Through electrostatic repulsion between the silver nanoparticles, the surface charge has a significant impact on the stability of the particles in suspension [31]. Additionally, the potential capping of the bioactive organic components found in the S. aromaticum extract contributes to the negative charge on the silver nanoparticles. The electrostatic repulsion between the silver nanoparticles is revealed by the high negative value of the particles, which shows that there was no agglomeration and the particles were stable. The silver particles’ tendency to interact with the negatively charged plasma protein is prevented by their negative charges. Due to their exposure to plasma proteins inside of a living organism, silver nanoparticles develop a protein corona that makes them stable in the biological medium [32].

The silver nanoparticles appeared semi-spherical in transmission electron microscopy images with an average size of 67 nm. While the particle size determined by DLS was slightly larger in size than the measurements obtained by TEM. The difference is due to the fact that TEM measures a limited number of silver particles, while DLS measures the hydrodynamic diameter of silver particles in solution or particles in dispersion, so DLS measures all particles in the dissolved sample [31]. The PDI can be recognized as a number computed from two parameters proportional to the correlation data despite the fact that it lacks dimensions and thus tends to have a value of 0. A polydispersive distribution of particles has a value of 1, whereas a monodispersive distribution has a value of 0. According to Danaei et al. [33] and Khorrami et al. [26], PDI values smaller than 0.05 indicate that the sample is monodispersive, whereas values larger than 0.7 km indicate a very wide distribution of silver particle size, and therefore it is not preferable to be analyzed by PDI technique.

Numerous studies have identified and quantified similar chemical compounds in the clove extract, Wadi [34] detected 58 compounds in S. aromaticum. Eugenol has the highest concentration, (53.24) major peaks were identified as eugenol, and is known to possess antimicrobial activity against many pathogens [35]. Similar findings confirmed the presence of 17 heterogeneous compounds, including eugenol (68.7–87.4 %), cyperene (20.5–7.2 %), phenethyl isovalerate (6.4–3.6 %), and cis-thujopsene were detected in both grounded and ungrounded seeds by GC–MS analysis (1.9–0.8 %) [36]. Research has investigated the pharmacological effects of S. aromaticum on various pathogenic parasites and microorganisms, such as Plasmodium, pathogenic bacteria, Theileria, Babesia, and hepatitis C viruses. Studies have reported that eugenol exhibits anti-inflammatory, anticancer, analgesic, antiseptic, antioxidant, antidepressant, antispasmodic, antifungal, antiviral, and antibacterial properties against several pathogens, including methicillin-resistant S. aureus and Staphylococcus epidermidis. Additionally, eugenol has been shown to protect against CCl4−induced liver toxicity and has demonstrated lethal effects on various parasites, including Fasciola gigantica, Giardia lamblia, S. mansoni, and Haemonchus contortus. This review explores the phytochemical composition and biological activities of clove extracts and essential oil, focusing on the primary active compound, eugenol [37]. Eugenol is rapidly absorbed and metabolized in the liver when ingested, and 95 % of the dose is excreted within 24 h. Acute toxicity is low by the oral route, with LD50 values ranging from 1,190–3,000 mg/kg-day. High oral doses of eugenol are acutely toxic to the liver in dogs and rats. Eugenol is a mild to moderate eye and skin irritant depending on the formulation. Subchronic toxicity is low, with NOAELs ranging from 900–>2,000 mg/kg-day. No data are available on endocrine disruption, reproductive, developmental, neurological, and immunotoxicity [37].

Previous studies focused solely on the bioactivity of silver nanoparticles (AgNPs) synthesized by S. aromaticum extract against various microorganisms, including bacteria such as E. coli, Staphylococcus aureus and Klebsiella pneumonia as well as fungi like Trichophyton rubrum [38]. These investigations demonstrated that AgNPs from S. aromaticum possess strong antibacterial and antifungal effects. However, the precise mechanisms behind the antimicrobial and antiparasitic actions of silver nanoparticles remain unclear. Some proposed mechanisms include the release of free silver ions that damage DNA, leading to disruptions in adenosine triphosphate production and the generation of reactive oxygen species. Another theory suggests that the adhesion of silver nanoparticles to affected cells disrupts the cell membrane, affecting its permeability. Additionally, silver nanoparticles are capable of penetrating target cells [39].

Numerous studies have sought to identify a potent and effective alternative to Praziquantel (PZQ) using extracts from medicinal plants. Abou El-Nour and Fadladdin [40] studied the impact of aqueous extracts from Coriandrum sativum, Piper nigrum, and Zingiber officinalis on S. mansoni. Additionally, Fadladdin [41] investigated the effects of aqueous extracts from Ziziphus spina-christi, Origanum majorana, and Salvia fructicosa on S. haematobium. Several other studies have explored the potential of metal nanoparticles as alternative treatments to PZQ. For instance, Khalil [42] utilized iron nanoparticles against S. mansoni and its intermediate host, Biomphalaria alexandrina, observing effects at a concentration of 30 μg/ml. Abou El-Nour [43] also assessed the effectiveness of copper oxide nanoparticles on S. mansoni and S. haematobium, achieving results at concentrations as low as 1.25 μg/ml. Dkhil [44] studied gold and selenium nanoparticles in vivo and found a significant impact on S. mansoni, which contributed to improved public health in murine models. To manage the intermediate host of S. mansoni, Moustafa [45] applied gold and silver particles against the snail B. alexandrina and its infectious cercariae, noting a mortality rate of 50 μg/ml for silver and 100 μg/ml for gold. Hamdan [46] compared the effects of green and chemical silver nanoparticles on S. mansoni, finding concentrations of 100 and 80 μg/ml to be the most effective in eliminating the worms. Eldera [47] studied biosynthesized zinc oxide nanoparticles by O. majorana aqueous leaves extracts against S. mansoni.

Xiao [48] found that exposing adult Schistosoma worms to 1–3 g/ml of PZQ resulted in immediate spastic paralysis in vitro. Furthermore, adult male worms exhibited surface blebbing on their tegument. In 2004, Pica-Mattoccia and Cioli [49] replicated this experiment using different concentrations of 0.1 and 1 g/ml, observing the same effects, which ultimately led to the death of all the schistosomes. These findings align closely with the effects observed with silver nanoparticles in the current study.

Although the initial side effects of the drug included a rapid influx of calcium into the parasite, resulting in calcium-dependent muscle contraction and paralysis [50], the proposed mechanism of PZQ suggests that the drug integrates into the membrane, causing a lipid phase transition and destabilizing the membrane. This theory indicates that Praziquantel interacts with the outer membrane of the tegument, leading to significant damage. Given that the silver nanoparticles synthesized from plant extracts produced similar effects to PZQ, it was anticipated that their mode of action would be the same [51].

It interacts between the host’s immune system and the outer surface of S. haematobium, which acts as an interface between the worms and their surrounding environment while also serving as a protective barrier. This tissue possesses various characteristics and functions, making it a primary target for anti-schistosomal drugs. Numerous studies have demonstrated the in vitro effects on the ultrastructural features of adult worms. Iron nanoparticles [42], copper oxide nanoparticles [43], and silver nanoparticles [46] exhibited similar effects to those observed with the current silver nanoparticles. These effects included loss of spines on the oral and ventral suckers, damage to spines, sloughing, formation of blebs, and significant oedematous areas. Several studies have investigated the effects of plant extracts on the worm’s surface [40], 45], 52], 53]. Ultrastructural assessments are typically conducted on male schistosome worms, as they are continuously exposed to the host’s micro-environment, whereas females are mostly confined within the male’s gynecological duct, making them less exposed. Additionally, males have numerous tubercles on their surface, making any changes in soft tissues more apparent.

Severe schistosomiasis can significantly disrupt liver function by altering its structure. The disease leads to varying levels of liver damage and immune responses that can progress to fibrosis. Many treatments for schistosomiasis aim to reduce, delay, or eliminate liver fibrosis, particularly in the chronic stages of the illness. Treatment also focuses on decreasing egg production and preventing schistosomes from depositing eggs in the liver. The current study indicated that chronic granulomatosis has a notable impact on liver parenchyma. Granulomas, which form due to the accumulation of schistosome eggs in fibrotic regions, contain a variety of chronic inflammatory cells, including plasma cells, lymphocytes, epithelial cells, eosinophils, and macrophages [54].

Overall, silver nanoparticles synthesized by S. aromaticum were found to be highly effective against schistosomiasis. This effectiveness was demonstrated by a reduction in both the size and number of granulomas, as well as a decrease in the number of eggs in the tissues of treated hamsters [41]. The therapeutic efficacy of these silver nanoparticles is attributed to the biologically active compounds present in the S. aromaticum extract, which includes various phytochemicals such as flavonoids, saponins, tannins, carbohydrates, quinones, alkaloids, and a phenolic glycoside/cardinolide [38]. Importantly, the proteins in the extract have immunomodulatory effects that lower Th2 cytokine production (linked to granuloma formation) while increasing Th1 cytokine production (associated with resistance to granuloma formation). This shift boosts the expression of IL-6, IL-1β, TNF-α, and iNOS, leading to enhanced neutrophil production and targeted leukocyte activity at sites of infection. Additionally, these proteins exhibit anti-edematogenic properties, normalize oxidative stress markers, and help maintain the structural integrity of the liver, spleen, and intestines [55]. These combined effects contribute to the observed reductions in granuloma size and number, explaining the significant improvements in the tissues of hamsters treated with silver nanoparticles.

The positive effects of silver nanoparticles synthesized by S. aromaticum, when mixed with PZQ in both in vitro and in vivo studies, have been demonstrated in numerous previous investigations [46], [56], [57], [58]. The key outcomes included an enhancement in the effectiveness of PZQ in laboratory tests. Additionally, in the groups treated with the nanoparticle mixture, there was a significant reduction in both the number and size of granulomas, as noted in this study.

6 Conclusions

The experimental data showed that treatment with green silver nanoparticles led to more severe visible changes in the teguments of adult S. haematobium worms and also caused significant damage to the oral and ventral suckers compared to those treated with PZQ alone. In in vivo experiments, green silver nanoparticles exhibited anti-schistosomal effects, in addition to their enhanced potency in augmenting the therapeutic effect of PZQ in the infected hamster model. The use of green silver nanoparticles led to greater reductions in the size/number of granulomas. This study supports the general trend of using safe, economical, and easy-to-prepare green nanoparticles in the treatment of fluke diseases. Mixing green silver nanoparticles with the traditional drug (PZQ) led to an improved effectiveness in killing adult worms. Silver may become a promising new candidate for developing new treatments for schistosomiasis.

Corresponding author: Samah S. Eldera, Physics Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia; and Physics Department, Faculty of Science, Al-Azhar University, Nasr City, Cairo, Egypt, E-mail:

Funding source: Deanship of Scientific Research (DSR) at King Abdulaziz University, Jeddah, Saudi Arabia

Award Identifier / Grant number: G: 144-247-1443

Acknowledgments

This project was funded by the Deanship of Scientific Research (DSR) at King Abdulaziz University, Jeddah, under grant No. (G: 144-247-1443). The authors, therefore, acknowledge with thanks DSR for technical and financial support.

  1. Funding information: The funding support for the research of this study was granted by the Deanship of Scientific Research (DSR) at King Abdulaziz University, Jeddah, Saudi Arabia (grant No. G: 144-247-1443).

  2. Author contribution: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Conflict of interest: The authors state no conflict of interest.

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

  5. Ethical approval: The in vivo research protocol followed the guidelines of the National Institutes of Health (NIH) Guide for Care and Use of Laboratory Animals (approval registration No. 16-257).

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Received: 2024-10-24
Accepted: 2025-09-27
Published Online: 2025-11-20

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

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

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https://doi.org/10.1515/rams-2025-0166
Keywords for this article
silver nanoparticles; green synthesis; Syzygium aromaticum; Schistosoma haematobium
Creative Commons
BY 4.0

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  71. Bioactive potential of marine Aspergillus niger AMG31: Metabolite profiling and green synthesis of copper/zinc oxide nanocomposites – An insight into biomedical applications
  72. Preparation of geopolymer cementitious materials by combining industrial waste and municipal dewatering sludge: Stabilization, microscopic analysis and water seepage
  73. Seismic behavior and shear capacity calculation of a new type of self-centering steel-concrete composite joint
  74. Sustainable utilization of aluminum waste in geopolymer concrete: Influence of alkaline activation on microstructure and mechanical properties
  75. Optimization of oil palm boiler ash waste and zinc oxide as antibacterial fabric coating
  76. Tailoring ZX30 alloy’s microstructural evolution, electrochemical and mechanical behavior via ECAP processing parameters
  77. Comparative study on the effect of natural and synthetic fibers on the production of sustainable concrete
  78. Microemulsion synthesis of zinc-containing mesoporous bioactive silicate glass nanoparticles: In vitro bioactivity and drug release studies
  79. On the interaction of shear bands with nanoparticles in ZrCu-based metallic glass: In situ TEM investigation
  80. Developing low carbon molybdenum tailing self-consolidating concrete: Workability, shrinkage, strength, and pore structure
  81. Experimental and computational analyses of eco-friendly concrete using recycled crushed brick
  82. High-performance WC–Co coatings via HVOF: Mechanical properties of steel surfaces
  83. Mechanical properties and fatigue analysis of rubber concrete under uniaxial compression modified by a combination of mineral admixture
  84. Experimental study of flexural performance of solid wood beams strengthened with CFRP fibers
  85. Eco-friendly green synthesis of silver nanoparticles with Syzygium aromaticum extract: characterization and evaluation against Schistosoma haematobium
  86. Predictive modeling assessment of advanced concrete materials incorporating plastic waste as sand replacement
  87. Self-compacting mortar overlays using expanded polystyrene beads for thermal performance and energy efficiency in buildings
  88. Enhancing frost resistance of alkali-activated slag concrete using surfactants: sodium dodecyl sulfate, sodium abietate, and triterpenoid saponins
  89. Equation-driven strength prediction of GGBS concrete: a symbolic machine learning approach for sustainable development
  90. Empowering 3D printed concrete: discovering the impact of steel fiber reinforcement on mechanical performance
  91. Advanced hybrid machine learning models for estimating chloride penetration resistance of concrete structures for durability assessment: optimization and hyperparameter tuning
  92. Influence of diamine structure on the properties of colorless and transparent polyimides
  93. Post-heating strength prediction in concrete with Wadi Gyada Alkharj fine aggregate using thermal conductivity and ultrasonic pulse velocity
  94. Experimental and RSM-based optimization of sustainable concrete properties using glass powder and rubber fine aggregates as partial replacements
  95. Special Issue on Recent Advancement in Low-carbon Cement-based Materials - Part II
  96. Investigating the effect of locally available volcanic ash on mechanical and microstructure properties of concrete
  97. Flexural performance evaluation using computational tools for plastic-derived mortar modified with blends of industrial waste powders
  98. Foamed geopolymers as low carbon materials for fire-resistant and lightweight applications in construction: A review
  99. Autogenous shrinkage of cementitious composites incorporating red mud
  100. Mechanical, durability, and microstructure analysis of concrete made with metakaolin and copper slag for sustainable construction
  101. Special Issue on AI-Driven Advances for Nano-Enhanced Sustainable Construction Materials
  102. Advanced explainable models for strength evaluation of self-compacting concrete modified with supplementary glass and marble powders
  103. Analyzing the viability of agro-waste for sustainable concrete: Expression-based formulation and validation of predictive models for strength performance
  104. Special Issue on Advanced Materials for Energy Storage and Conversion
  105. Innovative optimization of seashell ash-based lightweight foamed concrete: Enhancing physicomechanical properties through ANN-GA hybrid approach
  106. Production of novel reinforcing rods of waste polyester, polypropylene, and cotton as alternatives to reinforcement steel rods
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