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Biosynthesis of silver and gold nanoparticles using Annona squamosa seed extract: Insights into antibacterial and anticancer activities

  • Authors:
    • Firli Rahmah Primula Dewi
    • Aulia Umi Rohmatika
    • Almando Geraldi
    • A'Liyatur Rosyidah
    • Arniza Khairani Mohd Jamil
    • Turan Demircan
    • Versa Rachmania Hajar
    • Ameliora Clareista Elfentiana
  • View Affiliations / Copyright

    Affiliations: Department of Biology, Faculty of Science and Technology, Universitas Airlangga, Surabaya, East Java 60115, Indonesia, Research Center for Vaccine and Drugs, Research Organization for Health, National Research and Innovation Agency (BRIN), Bogor 16911, Indonesia, Department of Chemistry, Faculty of Science, Universiti Malaya, Kuala Lumpur 50603, Malaysia, Department of Medical Biology, School of Medicine, İzmir Bakırçay University, İzmir 35665, Turkey
    Copyright: © Dewi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 104
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    Published online on: July 9, 2026
       https://doi.org/10.3892/br.2026.2177
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Abstract

This study reports on the green synthesis of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) using Annona squamosa seed extract and the evaluation of their antibacterial and anticancer activities. The synthesized NPs were characterized by UV‑Vis spectroscopy, transmission electron microscopy, particle size and size distribution analysis, X‑ray diffraction, Fourier‑Transform Infrared Spectroscopy and Inductively Coupled Plasma‑Optical Emission Spectrometry. Characterization results confirmed the successful synthesis of spherical, crystalline NPs with mean hydrodynamic diameters of AgNPs and AuNPs determined as 40.60 and 72.65 nm, respectively. Antibacterial activity was assessed against Staphylococcus aureus and Escherichia coli, while anticancer activity was evaluated using MCF‑7 breast cancer cells and 293 cells. AgNPs exhibited significant antibacterial activity against both bacterial species, showing bacteriostatic activity against S. aureus and bactericidal activity against E. coli. By contrast, AuNPs showed no detectable antibacterial effect. AgNPs also demonstrated selective anticancer activity, inhibiting MCF‑7 cell proliferation in a dose‑ and time‑dependent manner with IC50 values of 106.704 and 79.519 µg/ml after 48 and 72 h of treatment, respectively, while exhibiting minimal toxicity toward 293 cells. Furthermore, AgNPs modulated the expression of several cancer‑related genes, suppressed oncogenic markers and induced apoptosis through activation of p53‑associated pathways, as evidenced by increased apoptotic cell populations and an elevated BAX/BCL2 ratio. Conversely, AuNPs displayed limited biological activity under the tested conditions. These findings highlight the potential of A. squamosa‑mediated AgNPs as promising antibacterial and selective anticancer agents for future biomedical applications.
View Figures

Figure 1

Synthesis of NPs using A.
squamosa seeds extract and their characterization. (A) The
observed color changes during the synthesis of AgNPs. (B) UV-Vis
spectrophotometric analysis of the synthesized AgNPs. (C) TEM
visualization of AgNPs synthesized using A. squamosa seed
extract showing predominantly spherical NPs with crystalline
lattice fringes (scale bars, 20 and 5 nm). (D) The color changes
observed during the synthesis of AuNPs. (E) UV-Vis spectroscopic
analysis of the synthesized AuNPs. (F) TEM visualization of AuNPs
synthesized using A. squamosa seed extract exhibiting
spherical to quasi-spherical morphology and highly ordered
crystalline structures (scale bars, 20 and 5 nm). AgNPs, silver
nanoparticles; Au, gold; TEM, transmission electron microscopy;
A. squamosa, Annona squamosa.

Figure 2

Size distribution XRD patterns
confirm the crystalline structures of the NPs. (A) PSA result of
AgNPs. (B) NP size analysis of AgNPs using ViewSizer 3000. (C) XRD
patterns of AgNPs. (D) PSA result analysis of AuNPs (E). NP size
analysis of AuNPs using ViewSizer 3000. (F) XRD patterns of AuNPs.
(G) The fourier transform infrared (FT-IR) spectra of A.
squamosa seed extract, AgNPs and AuNPs. AgNPs, silver
nanoparticles; AuNPs, gold nanoparticles; a.u., absorption units;
XRD, X-ray diffraction; PSA, particle size analyzer; PDI,
polydispersity index.

Figure 3

Sterility and antibacterial activity
of the NPs. (A) Sterility test of AgNPs. (B) The antibacterial
activity of AgNPs and AuNPs against S. aureus and E.
coli at concentrations of 10, 25, 50 and 100 mg/ml, with a 24-h
incubation period (scale bars, 6 mm). CTL, control; SQ, A.
squamosa seed extract; AgNPs, silver nanoparticles; AuNPs, gold
nanoparticles.

Figure 4

Effects of Annona squamosa
seed extract and silver- or gold-based formulations on the
viability of MCF-7 and 293 cells. (A) Viability of MCF-7 cells
following treatment with SQ for 24, 48 and 72 h. (B) Viability of
MCF-7 cells following treatment with AuNPs synthesized using A.
squamosa seed extract at various concentrations for 24, 48 and
72 h. (C) Viability of MCF-7 cells following treatment with
AgNO3 at various concentrations for 24 h. (D) Viability
of MCF-7 cells following treatment with AgNPs synthesized using
A. squamosa seed extract at various concentrations for 24,
48 and 72 h. (E) Viability of 293 cells following treatment with
AgNO3 at various concentrations for 72 h. (F) Viability
of 293 cells following treatment with AgNPs at various
concentrations for 24 and 72 h. Data are presented as the mean ±
standard deviation from three independent experiments (n=3).
Statistical significance was determined using one-way ANOVA, with
comparisons made against the untreated control group. Asterisks
indicate significant differences: *P<0.05,
**P<0.01 and ****P<0.0001 vs. CTL. CTL,
control; SQ, A. squamosa seed extract; AgNPs, silver
nanoparticles; AuNPs, gold nanoparticles.

Figure 5

IC50 of silver
nanoparticles prepared with A. squamosa seeds extract
against the MCF7 cell line after (A) 24, (B) 48 and (C) 72 h of
treatment.

Figure 6

AgNPs treatment alters cell cycle
phase and several oncogene expressions in MCF-7 cells. (A) The
nuclear size and nuclear area of the MCF-7 cells following
treatment with 100 µg/ml of AgNPs for 24 h. (B) The DNA content
represents cell cycle phase of MCF-7 cells following treatment with
100 µg/ml of AgNPs for 24 h. (C) Relative mRNA levels of MYC, (D)
CCND1, (E) HER-2, (F) COX-2 and (G) CTNND1 in MCF-7 cells following
treatment with AgNPs (100 µg/ml) for 48 h. Data are presented as
mean ± standard deviation of three independent replicates (n=3).
Statistical analysis was performed using one-way ANOVA. Asterisks
indicate statistically significant differences compared with the
control group: *P<0.05, **P<0.01,
***P<0.001 and ****P<0.0001 vs. CTL.
ns, no significance; CTL, control; MYC, c-Myc; CCND1, cyclin D1;
HER2, human epidermal growth factor receptor-2; COX-2,
cyclooxygenase-2, CTNND1, catenin delta-1; AgNPs, silver
nanoparticles.

Figure 7

AgNP treatment induces apoptosis and
modulates the expression of apoptosis-related genes in MCF-7 cells.
(A) Representative flow cytometry dot plots of Annexin
V-FITC-stained MCF-7 cells showing apoptotic populations in the
untreated control and AgNP-treated (100 µg/ml, 48 h) groups. (B)
Quantification of apoptotic cells demonstrating a significant
increase in apoptosis following AgNP treatment compared with the
control group, as determined by Student's t-test. (C) Relative
expression levels of apoptosis-related genes following AgNP
exposure. AgNP treatment significantly upregulated p53, BAX and
BAX/BCL2 ratio, while significantly downregulating PUMA expression
compared with the control. Gene expression data were analyzed using
two-way ANOVA and are presented as the mean ± standard deviation
from three independent experiments (n=3). Statistical significance
is indicated as *P<0.05 and ***P<0.001
vs. control. ns, not significant; AgNPs, silver nanoparticles;
TP53, tumor protein P53; BAX, BCL2 associated X; BBC3/PUMA, BCL2
binding component 3; BCL2, BCL2 apoptosis regulator.
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Copy and paste a formatted citation
Spandidos Publications style
Dewi FR, Rohmatika AU, Geraldi A, Rosyidah A, Mohd Jamil AK, Demircan T, Hajar VR and Elfentiana AC: Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities. Biomed Rep 25: 104, 2026.
APA
Dewi, F.R., Rohmatika, A.U., Geraldi, A., Rosyidah, A., Mohd Jamil, A.K., Demircan, T. ... Elfentiana, A.C. (2026). Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities. Biomedical Reports, 25, 104. https://doi.org/10.3892/br.2026.2177
MLA
Dewi, F. R., Rohmatika, A. U., Geraldi, A., Rosyidah, A., Mohd Jamil, A. K., Demircan, T., Hajar, V. R., Elfentiana, A. C."Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities". Biomedical Reports 25.3 (2026): 104.
Chicago
Dewi, F. R., Rohmatika, A. U., Geraldi, A., Rosyidah, A., Mohd Jamil, A. K., Demircan, T., Hajar, V. R., Elfentiana, A. C."Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities". Biomedical Reports 25, no. 3 (2026): 104. https://doi.org/10.3892/br.2026.2177
Copy and paste a formatted citation
x
Spandidos Publications style
Dewi FR, Rohmatika AU, Geraldi A, Rosyidah A, Mohd Jamil AK, Demircan T, Hajar VR and Elfentiana AC: Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities. Biomed Rep 25: 104, 2026.
APA
Dewi, F.R., Rohmatika, A.U., Geraldi, A., Rosyidah, A., Mohd Jamil, A.K., Demircan, T. ... Elfentiana, A.C. (2026). Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities. Biomedical Reports, 25, 104. https://doi.org/10.3892/br.2026.2177
MLA
Dewi, F. R., Rohmatika, A. U., Geraldi, A., Rosyidah, A., Mohd Jamil, A. K., Demircan, T., Hajar, V. R., Elfentiana, A. C."Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities". Biomedical Reports 25.3 (2026): 104.
Chicago
Dewi, F. R., Rohmatika, A. U., Geraldi, A., Rosyidah, A., Mohd Jamil, A. K., Demircan, T., Hajar, V. R., Elfentiana, A. C."Biosynthesis of silver and gold nanoparticles using&nbsp;<em>Annona squamosa</em> seed extract: Insights into antibacterial and anticancer activities". Biomedical Reports 25, no. 3 (2026): 104. https://doi.org/10.3892/br.2026.2177
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