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Zinah Ayyed Habeeb1
, Sanaa Khudhur Jameel1and Mais Emad Ahmed2*
1Medical Microbiology Department, College of medicine, AL-Iraqia University, Baghdad, Iraq.
2Department of Biology, College of Science, University of Baghdad, Jadriya, Baghdad, Iraq.
Corresponding Author E-mail:mais.emad@sc.uobaghdad.edu.iq
DOI : https://dx.doi.org/10.13005/bpj/3139
Abstract
Studies on the functional characteristics of Se nanoparticles (SeNP) have become increasingly popular in recent decades because of their strong selectivity, stability, and high biocompatibility. Numerous studies demonstrate SeNP's anticarcinogenic properties. In order to defeat bacteria that are resistant to antibiotics, new strategies need to be developed. A subclass of antimicrobial peptides known as selenium nanoparticles (NPs) can be used either in place of or in addition to well-known antibiotics. SeNPs have been investigated with the Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), zeta potential, and transmission electron microscopy (TEM)., and UV-visible analysis. The antibacterial qualities of SeNPs were confirmed by the generated nanoparticles showed increased antimicrobial activity at different concertation (1000-64) µg/ml In the good diffusion assay, the NPs efficiently suppressed the growth of susceptible SeNPs have zones of inhibition between 5-19 mm against Gram-Negative while against Gram Positive bacteria Staphylococcus epidermidis and Staphylococcus aureus bacteria have zones of inhibition between 6-17 mm , respectively. Our study has shown that combining various antimicrobials with different mechanisms of action led to an extensive variety of inhibition and raised efficacy, particularly against germs that are resistant to several medications. The best anti- human breast cancer cell line MCF7 properties of nanocomposite against the above cell lines was in the case of MCF7 cell line. According to the above findings, the SeNPs may be administrated for the treatment of several types of human breast cancer in humans. Ovarian cancer had increased nanoparticle sensitivity. At a concentration of 25 μL/mL, SeNPs demonstrated excellent cytotoxicity and anticancer activity, making them suitable for use as an anticancer agent.The physicochemical characteristics, experimental setup, and concentrations of SeNPs exploited in this study must all be carefully stated.
Keywords
Ascorbic Acid; Selenium Nanoparticles; X-ray Diffraction; Cell lines (MCF7); GI50 value
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| Copy the following to cite this article: Habeeb Z. A, Jameel S. K, Ahmed M. E. Synthesis, Characterization, and Cytotoxic Evaluation of Selenium Nanoparticles. Biomed Pharmacol J 2025;18(1). |
| Copy the following to cite this URL: Habeeb Z. A, Jameel S. K, Ahmed M. E. Synthesis, Characterization, and Cytotoxic Evaluation of Selenium Nanoparticles. Biomed Pharmacol J 2025;18(1). Available from: https://bit.ly/4ky3wiJ |
Introduction
The field of theranostics, which integrates therapy and diagnostics, has advanced significantly due to thanks to developments in nanotechnology, pharmaceutical science, and healthcare. With selenium nanoparticles (SeNPs), which are growing into more and more effective theranostic agents, this is especially so Some studies.1 Antioxidant, anti-inflammatory, anticancer, antibacterial, antidiabetic, wound healing, and cytoprotective therapies are just a few of the areas in which SeNPs have demonstrated great promise. The review emphasizes SeNPs’ versatility and biocompatibility, which are essential for sophisticated disease monitoring, detection, and individualized care. Particular attention is given to developments in green synthesis methods, highlighting their economical and environmentally beneficial uses in biosensing, imaging, diagnostics, and therapy .2 Because of its unique bioactivities in nanoforms, selenium (Se), a crucial trace element, stands out among these nanoparticles. Through oxidoreductase activity, a vital enzymatic mechanism, selenium is integrated into selenium-containing proteins like selenocysteine (Sec)”like selenocysteine (Sec), which are crucial for preserving physiological redox equilibrium. Devices based on selenium have been created in the field of medication delivery to deliver drugs to particular areas Researchers like Karthik et al .,3 The production of selenium nanoparticles (SeNPs) via biological synthesis techniques and their uses in a variety of industries. The environmental-friendly characteristics of plant-based synthesis, which uses ingredients like aloe vera leaf extracts, has drawn research. Other research have employed various species, such as the seeds of Ganoderma lucidum spore oil and Catathelasmaventricosum polysaccharides, for SeNP synthesis, exhibiting a variety of methods. Aloe vera extracts, which are abundant in secondary metabolites, work as natural reductants and stabilizers in SeNP creation Some studies,4 Selenium has a limited range of therapeutic and dangerous dosages, and even subtoxic concentrations can have negative health effects, despite its critical role in proper biological functioning. Because of their high bioavailability and low toxicity, selenium nanoparticles (NPs) efficiently enhance the delivery and absorption of this trace element without producing toxicity Another investigation. 5 There is a great deal of typological variety in the contentious link between exposure dose and the negative effects of selenium nanoparticles. To improve the risk assessment of these molecules, more research is required to elucidate the absorption, metabolism, and long-term toxicity of selenium nanoparticles Some studies. 6
We also go over the latest research on the potential of a number of biological materials for the synthesis of selenium nanoparticles. The physical characteristics of the product (size, shape, and stability) will be discussed together with the precursor, extract, procedure, time, temperature, and other synthesis requirements. Future researchers can more accurately synthesize SeNPs and use them in desired applications thanks to the synergy of SeNP synthesis via many Somestudies.7 Thus, the purpose of this study was to create antibacterial composite resins using crystalline selenium nanoparticles (SeNPs) and evaluate the composite resin’s mechanical, physical, and antibacterial qualities following SeNP integration.
Material and Methods
SeNPs’ characterization and environmentally friendly manufacture Selenium nanoparticles (SeNPs) were produced by a reduction reaction between sodium selenite and ascorbic acid. 50 mM sodium selenite and stabilizing agents were combined with magnetic stirring for five minutes to produce stabilized SeNPs. The solutions were then gradually supplemented with 1 weight percent ascorbic acid in deionized water. To attain total reduction, the reaction mixtures were shaken for half an hour studies are needed.8
Characterization of SeNPs
Characterization is essential in order to fully understand the distinctive properties of nanoparticles. The methods that followed have been utilized to determine the nanoparticles’ sizes and shapes: The evaluation of surface structure, particle size distribution, particle or crystal effect, nanoparticle aggregation, surface functionalization, and single-particle assessments can be seen using Field Emission Scanning Electron Microscopy (FE-SEM). A device used in materials research to ascertain a material’s crystallographic organization is called an X-ray diffraction machine (XRD).. Fourier-transform infrared spectroscopy (FT-IR, RX I, PerkinElmer, Inc., USA) at a resolution of 4 cm1 and Zeta Potential (ZP) and energy dispersive X-ray (EDX) were used to analyze the functional groups of produced nanostructures in the 4000–400 nm range Auther Salman et al,. 9
MIC determination
The lowest concentration of SeNPs that restricts bacterial growth was identified (MIC) using Eppendorf tubes as the medium and the broth microdilution technique. After the production in MHB, the microbial inoculum’s concentration was lowered to 100 UFC/μl. Two-fold serial dilutions of NPs (1000, 500, 250, 125, and 64 μg/μl) were produced using MHB in six tubes. The tubes were afterward incubated for a full day at 37° C. Following that, results were acquired by looking for indicators of bacterial growth in each tube, making it possible for an estimation of the subMIC Another investigation . 10
Testing the antimicrobial activity of SeNPs
Natural Products-Based Metallic Nanoparticles possess a variety of bioactivities, such as antimicrobial properties. The microorganism’s resistance to multiple drugs and its distinct cell membrane composition limit its bioactivity. Mueller Hinton Agar was used for the Agar Well Diffusion assay. Sterilized inoculating loop or swab were used to spread the test isolates Clinical isolate included gram-negative and gram-positive were obtained from the College of Medicine at Iraqi University were examined. onto Muller-Hinton Agar plates. A sterile cork borer was used to create 5 wells 5 mm, which were then filled with 80 µl of varied concentrations depends on microdilution technique below. Then subsequently incubated for 24 hours at 37 ° C, after which the zones of inhibition were determined the following day Another investigation. 11
Cytotoxicity assay
The cytotoxic effects were determined with an in vitro 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) test. Seven billion cells were introduced in each well of a 96-well plate and incubated overnight to ensure adhesion between cells. After that, cell lines (MCF7) were treated to substances at progressively higher SeNPs concentrations (6.25–100 μg/ml); three replicate wells were employed for each treatment. After 24 hours of incubation, the media was taken out of the plate, and each well received 20 μl of MTT solution (5 mg/ml) from Shanghai Macklin Biochemical Co., Ltd. The wells were then incubated for 3 hours at 37˚C in the dark. After adding 50 μl of DMSO (Bio Basic Inc.), To dissolve the MTT, it was rattled for a total of ten minutes. A BioTek Instruments, Inc. microplate reader was used for identifying absorbance at 490 nm. The following formula was used to determine the number of living cells from the raw fluorescence data.
Where the absorbance is denoted by “A.” GraphPad Prism software version 6 (Dotmatics) was used to create the dose-response curve, and the same curve was used to calculate the growth inhibitory concentration (GI50), which lowers viability by 50% Some studies,12
Results
Synthesis of SeNPs
This medium’s color shift, which turned red once Se2O3 had been reduced to Se0, was the first hint of the synthesis of the SeNPs. After the powdered SeNPs nanoparticles had been produced at 120°C, they were described, and the formation of a white precipitate showed that the nanoparticles were formed. The precipitate turns white after centrifugation. After microwave drying, we were presented with a beautiful white powder.
Characterization of SeNPs
UV–Vis Spectral Analysis and Zeta Potential Analysis
The reduction to Se0 was demonstrated by the UV-Vis spectroscopic research studies. The acquisition of a strong absorption signal at 288 nm validated the production of SeNPs while Na2SeO3. The peak attained 298 nm as the Na2SeO3 concentration grew (Fig. 1a). On the other hand, the zeta potential of He10214 was -51.2 mV, while that of the SeNPs produced by Si10198 was 60.6 mV (Fig. 1b).
Atomic Force Microscopy (AFM) analysis and X-ray analysis
AFM analysis of SeNPs was performed with identify and characterize distributions of nanoparticles. The nanoparticales microstructure was investigated utilizing atomic force microscopy (AFM). Nanoparticles were discovered to have irregular and triangle cluster morphologies, ranging between 0.25 and 23.33 nm in height and 13 to 177 nm in diameter (as verified by a particle size analyzer) according to the three-dimensional AFM. (Fig. 2a). XRD assessing further verified the chemical production of SeNPs (Fig. 2b). The strongest peak was seen at 54 cps, and the peak value for diffraction was roughly 29.5 (2θ).
 |
Figure 1: a) UV analysis b) Zeta potential measurement of the SeNPs of synthesized |
 |
Figure 2 : a) Atomic Force Microscopy analysis b) X-ray analysis of SeNPs |
Field emission Scanning Electron Microscope (FESEM) and EDX analysis
Images of SeNPs under an electron microscope appear in Fig. 3.a. The typical lengths of these spherical particles in 6 and 8 mM of the synthesized Na2SeO3 are around 50–100 nm and 30–100 nm, respective. The spherical shape of the biosynthesised SeNPs was confirmed by FESEM images.The unchanged chitosan nanoparticles are composed of clusters of particles that vary in size from 10 to 33 nm. The synthesized SeNPs’ EDX investigation, as shown in Fig. 3.b.
 |
Figure 3: a) FESEM Images b) EDX analysis confirming selenium element |
FTIR Analysis
High absorption can be noticed in the FTIR spectra of the unaltered (CH/GL) specimen film at 3293 cm−1 (-OH stretching), 2925 cm−1 (amide A’s N-H stretching), 1549 cm−1 (C-C, C bending), 1410 cm−1 (C-O broadening), and 1640 cm−1 (C=O stresses). The carbonyl component (-C=O) of chitosan and the C-H group of gelatin interact in the FTIR a spectrum of the blended composite films (Figure 4).
 |
Figure 4: Fourier-transform infrared spectroscopy (FTIR) spectra |
Antibacterial activity SeNPs
Because selenium nanoparticles has antimicrobial action both on its own and when metal nanoparticles bond to this substance, researchers are very interested in SeNPs nanoparticles. It has outstanding antibacterial qualities. Multiple explanations for selenium nanopartical antibacterial effects have been offered by scientists. The antibacterial properties of the targeted metal NPs against four Gram-Positive and negative bacteria were observed are shown Figures 5-a and 5- b respectively. The standard drug kanamycin’s zone of inhibition (measured in millimeters) against the Gram-negative bacteria Klebsiella pneumoniae and Escherichia coli were at concentration 1000 µg/ml found 17 mm and 18 mm respectively. On other staphylococcus aureus and staphylococcus epidermises at the same concertation range inhibition zone reached 8 and 17 mm at concentration 1000 µg /ml.
 |
Figure 5: Mean ± SD Zone of Inhibition in mm treated with SeNPs against a) gram-positive bacteria and b gram-negative at Different Concentrations between 1000, 500, 250, 125, and 62.5 μg/mL.: Standard Deviation, (n = 3) |
The cytotoxic effect of SeNPs nanoparticles on tumor cell lines
The cytotoxic activity of selenium nanoparticles on the human breast adenocarcinoma cell line MCF7 were studied as seen in Table 1 and Figure 6. The results obtained showed that the cell viability reduced as the concentration of SeNPs increased. The GI 50 value of SeNPs was about 24. 5 µg/ml. These NPs exhibited toxic effects against the MCF7 cell line, and at a concentration of 25 µg/ml, the cytotoxic effects reached 65.4%. SeNPs were tested for in vitro cytotoxicity against MCF7 cell lines at concentrations of 6.25–100 μg/ml; In agreement with our results, several evidence have indicated the cytotoxic potential of SeNPs on different cell lines.
Table 1: The cytotoxicity of SeNPs on cell line MCF7
| CV: Control vehicle
TR: Technical replicate |
|||||||
| MDA-MB231 | CV | 6.25 uM | 12.5 uM | 25 uM | 50 uM | 100 uM | |
| Compound 1 | Raw data | 0.361 | 0.254 | 0.308 | 0.201 | 0.241 | 0.136 |
| TR1 | % Survival | 100 | 70.36011 | 85.31856 | 55.67867 | 66.759 | 37.67313 |
 |
Figure 6: The cytotoxic effect of SeNPs on MCF7 cells |
Discussion
All three SeNPs have been determined to be nearly spherical by scanning electron microscopy (SEM) analysis, and the purified SeNPs’ EDAX microanalysis demonstrated substantial selenium absorption maxima at 1.37 (SeLa), 11.22 (SeKa), and 12.49 keV (SeKb). Additionally, the elemental compositions in various synthesised SeNPs changed Researchers like. 13 In biogenic SeNPs, for instance, the weight percentage of selenium was 9.26% for Bm-SeNPs(+) and 11.01% for Sm-SeNPs(~).However, ChSeNPs had a greater selenium proportion (31.61%). Additionally, the crystalline makeup of biogenic SeNPs, which were rich in C, O, P, and S, indicate the presence of biological polysaccharides close to the nanostructures. Based on the presence and quantity of C, O, and S, one might believe that the biogenic SeNPs cap is composed of proteins or enzymes, specific cellular residues, and phospholipids found in the membrane (P peaks) 14. Se-Ps’ electrical structure and, in turn, their ability to absorb and scatter light are influenced by their size,the UVvis spectrum is where these impacts are most apparent. This technique aids researchers in analyzing the interactions between NPs and light, yielding crucial data for comprehending NP size and quantum characteristics.
Relevant information regarding the size, shape, and composition of the particles are provided by this procedure. NPs’ small size causes quantized energy levels Another investigation. 15 Studies using UV-vis spectroscopy have demonstrated that Se-NPs absorb light differently due to their differing electrical configurations. To comprehend how light interacts with NPs, surface plasmon resonance data is crucial. Se-NPs have been verified by researchers based on peaks in the ultraviolet (UV) (200–400 nm) Studies by Instructions to Authors. 16 These Se-NPs have a 45–0 nm diameter and a spherical shape. Particle aggregation takes over during the creation of nanoparticles (NPs), concealing the reduction of precursor atoms and their initial nucleation Some studies,17 XRD may be used to discern distinguish various translucent forms of selenium, particularly the hexagonal and amorphous forms. Peak broadening in the XRD pattern, which is associated with the size of the crystalline structures in the NPs, can be used to estimate the average Se-NP particle size from XRD data. Furthermore, XRD can show which impurities are present in the caused Se-NPs and authenticate the chemical formula and stoichiometry of Se-NPs. Some studies, 18. Examining the cytotoxicity of green produced Se-NPs is essential for determining their appropriateness for biological applications. The green synthesis approach creates Se-NPs from natural and eco-friendly sources, like plant extracts, which are thought to be less hazardous than chemically produced Se-NPs Authors Puri et al ,.19 Some doses of Se have been shown to be detrimental. Consequently, it is thought that the size, structure, and dosage of Se nanoparticles affect their toxicity. Several procedures have demonstrated that biogenic Se-NPs are less potentially potentially dangerous to animals than sodium selenite. Bano et al. defined that low concentrations of Se-NPs are safe after analyzing the toxicological effects of Se-NPs in animals Another investigation. 20
Conclusions
All physiological functions depend on selenium, a necessary element that offers nutritional value. In order to prevent health abnormalities caused by selenium insufficiency, it is crucial to maintain a sufficient supply of selenium. Because nano-selenium is readily available, has low toxicity, SeNPs are an antioxidant, antibiofilm, antibacterial, and anticancer that is also involved in the dietary supplement. SeNPs are less harmful at the right dosage, according to numerous toxicity studies. Future studies on Se-NPs should concentrate on developing characterisation tools, looking for Se-NPs’ potential in targeted medication delivery systems and enhancing green synthesis techniques for scalability and sustainability. To guarantee safety, thorough toxicity and environmental effect analyses will be necessary, and in-
Acknowledgement
The authors would like to acknowledge the Departments of Biology, College of Science, University of Baghdad and Medical and Molecular Biotechnology Department, Biotechnology Research Center, Al-Nahrain University, for their great support in performing this work in their laboratories.
Funding Sources
The author(s) received no financial support for the research, authorship, and/or publication of this article.
Conflict of Interest-
The author(s) do not have any conflict of interest.
Data Availability Statement
This statement does not apply to this article.
Ethics Statement
This case report was conducted according to the Helsinki Declaration. The collection and evaluation of the data were confidential according to the health policy of the Iraqi Ministry of Health IN September 22-9-2024 Ref:CSEC/0922/0086
Informed Consent Statement
This study did not involve human participants, and therefore, informed consent was not required.
Clinical Trial Registration
This research does not involve any clinical trials
Author Contribution
- Zinah Ayyed Habeeb: writing—original draft methodology, investigation, and formal analysis.
- Sanaa Khudhur Jameel: writing—original draft methodology, investigation, and formal analysis.
- Mais. Emad. main concepts, data interpretation, supervision, and all reviewed the manuscript
References
- Nag S., Kar S., Mishra S. et al. Unveiling green synthesis and biomedical theranostic paradigms of selenium Nanoparticles (SeNPs)-A State-of-the-Art comprehensive update. International Journal of Pharmaceutics. 2024;124535-124542.
CrossRef - Waqar M. A. A comprehensive review on recent advancements in drug delivery via selenium nanoparticles. Journal of Drug Targeting.2024; 1-14.
CrossRef - Karthik K., Cheriyan B. V., Rajeshkumar S. et al. A review on selenium nanoparticles and their biomedical applications. Biomedical Technology.2024; 61-74.
CrossRef - Zhou S., Zhu H., Xiong P. et al. Spore oil-functionalized selenium nanoparticles protect pancreatic beta cells from palmitic acid-induced apoptosis via inhibition of oxidative stress-mediated apoptotic pathways. Antioxidants. 2023; 12(4); 840.
CrossRef - Ahmed, M. E., Sulaiman, G. M., Hasoon, B. A et al . Green Synthesis and Characterization of Apple Peel‐Derived Selenium Nanoparticles for Anti‐Fungal Activity and Effects of MexA Gene Expression on Efflux Pumps in Acinetobacter baumannii. Applied Organometallic Chemistry, 2024;7805-7811.
CrossRef - Ryabova, Y. V., Sutunkova, M. P., Minigalieva, I. A., et al. Toxicological effects of selenium nanoparticles in laboratory animals: A review. Journal of Applied Toxicology. 2024; 44(1): 4-16.
CrossRef - Sampath S, Sunderam V, Manjusha M, Dlamini Z et al. Selenium Nanoparticles: A Comprehensive Examination of Synthesis Techniques and Their Diverse Applications in Medical Research and Toxicology Studies. Molecules. 2024; 29(4): 801.
CrossRef - El-Deeb B, Al-Talhi A, Mostafa N and Abou-assy R. Biological synthesis and structural characterization of selenium nanoparticles and assessment of their antimicrobial properties. Am Sci Res J Eng Technol Sci. 2018; 45: 135–70 .
- Salman, F., and ME, A. M. N. A. The effect of selenium nanoparticles on the expression of MexB gene of Pseudomonas aeruginosa Isolated from wound and burn infections. Iraqi JMS. 2024; 22 (1): 79-92.
- Hassan, Z. J. S., Hamid, M. K., and Ahmed, M. E. Synthesized Zinc Oxide Nanoparticles by the Precipitation Method on Streptococcus spp. from Dental Carries and Cytotoxicity Assay. J. Drug Deliv. Technol.2022 ; 12, : 1327-1330
CrossRef - Ahmed, M. E., Hamza Faiq, N., Almutairi, H. H., & Alam, M. W. (2025). Biosynthesized ZnO-CuO Nanocomposite for Biofilm Formation of Proteus mirabilisupon LuxS Gene Expression. Inorganics, 13(2), 65. https://doi.org/10.3390/inorganics13020065
CrossRef - Mohammed Mahdi S., Fadhil Abbas A., M.E. Ahmed. Cytotoxicity of zinc oxide nanoparticles to lymphocytes using Enterococcus faecium bacteriocin and assessment of their antibacterial effects. Nanomed Res J, 2024; 9(2) :180-194.
- Filipović, N., Ušjak, D., Milenković, M. T ,et al . Comparative study of the antimicrobial activity of selenium nanoparticles with different surface chemistry and structure. Frontiers in bioengineering and biotechnology. 2021; 8:624621-624630.
CrossRef - Fawzi, F. H., and Ahmed, M. E. Green Synthesis and Characterization of Selenium Nanoparticles via Staphylococcus warneri Approach: Antimicrobial and on PhzM Pyocyanin Gene Expression in Pseudomonas aeruginosa. Plasmonics .2024 ; 1-17.
CrossRef - Kumar A, Shah SR, Jayeoye TJ, Kumar A, Parihar A, et al. Biogenic metallic nanoparticles: biomedical, analytical, food preservation, and applications in other consumable products. Front Nanotechnol .2023; 5: 1175149.
CrossRef - Joudeh N, Linke D. Nanoparticle classification, physicochemical properties, characterization, and applications: a comprehensive review for biologists. J Nanobiotechnol .2022;20(1): 262
CrossRef - Alagesan V, Venugopal S. Green synthesis of selenium nanopar- ticle using leaves extract of Withania somnifera and its biolog-ical applications and photocatalytic activities. BioNanoScience 2019;9 (1): 105-16.
CrossRef - Amin MA, Ismail MA, Badawy AA et al potency of fungal-fabricated selenium nanoparticles to improve The the growth performance of Helianthus annuus L. and control of cut-worm agrotis ipsilon. Catalysts 2021;11(12): 1551
CrossRef - Puri A, Mohite P, Ansari Y, Mukerjee N, Alharbi HM . Plant-de- rived selenium nanoparticles: investigating unique morphologies,enhancing therapeutic uses, and leading the way in tailored med-ical treatments. Mater Adv ,2024;5(9):3602-28.
CrossRef - Bano I, Skalickova S, Arbab S, Urbankova L, Horky P. Toxi- cological effects of nanoselenium in animals. J Anim Sci Biotechnol 2022;13(1): 72
CrossRef
