Abstract
The use of metal nanoparticles (NPs) conjugated with natural herbal molecules in biomedical applications has been growing. In this work, we synthesized Iron oxide NPs conjugated with thymol (Fe3O4@Glu-Thymol) and investigated their antibacterial and anticancer potentials. Physicochemical features of the NPs were studied by FT-IR, EDS-mapping, XRD, DLS, zeta potential, and electron microscopy. The antibacterial activity of the NPs against Pseudomonas aeruginosa and anticancer activity for breast cancer cells was investigated by broth microdilution and MTT and flow cytometry assays, respectively. The expression of apoptosis signaling genes in breast cancer cells that were treated with the NPs was studied by qPCR assay. The NPs were spherical, in a size range of 40–66 nm, without impurities, and with zeta potential and hydrodynamic size of − 23 mV and 185 nm, respectively. Moreover, the FT-IR and XRD assays confirmed the proper synthesis of Fe3O4 and conjugation with thymol. The minimum inhibitory concentration of the NPs for P. aeruginosa strains was 64–128 µg/mL. Our results showed that Fe3O4@Glu-Thymol was considerably more toxic for breast cancer cells than normal human cells and the 50% inhibitory concentration were 90.4 and 322 µg/mL, respectively. Upon treating breast cancer cells with the NPs the frequency of cell apoptosis increased by 18.9%. Also, the expression of the BAX and CASP8 genes in NPs treated cells significantly increased by 1.75 and 2.25 folds, respectively while the BCL-2 gene remained almost constant. This study reveals that Fe3O4@Glu-Thymol has considerable potential to be used in biomedical fields.
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References
Awasthi R, Roseblade A, Hansbro PM, Rathbone MJ, Dua K, Bebawy M (2018) Nanoparticles in cancer treatment: opportunities and obstacles. Curr Drug Targ 19(14):1696–1709. https://doi.org/10.2174/1389450119666180326122831
Bhalla Y, Gupta VK, Jaitak V (2013) Anticancer activity of essential oils: a review. J Sci Food Agric 93(15):3643–3653. https://doi.org/10.1002/jsfa.6267
Bigdeli R, Shahnazari M, Panahnejad E, Cohan RA, Dashbolaghi A, Asgary V (2019) Cytotoxic and apoptotic properties of silver chloride nanoparticles synthesized using Escherichia coli cell-free supernatant on human breast cancer MCF 7 cell line. Artif Cells Nanomed Biotechnol 47(1):1603–1609. https://doi.org/10.1080/21691401.2019.1604533
Birsoy K, Possemato R, Lorbeer FK, Bayraktar EC, Thiru P, Yucel B, Sabatini DM (2014) Metabolic determinants of cancer cell sensitivity to glucose limitation and biguanides. Nature 508(7494):108–112. https://doi.org/10.1038/nature13110
Brentnall M, Rodriguez-Menocal L, De Guevara RL, Cepero E, Boise LH (2013) Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis. BMC Cell Biol 14(1):1–9. https://doi.org/10.1186/1471-2121-14-32
Ezealigo US, Ezealigo BN, Aisida SO, Ezema FI (2021) Iron oxide nanoparticles in biological systems: antibacterial and toxicology perspective. JCIS Open 4:100027. https://doi.org/10.1016/j.jciso.2021.100027
Habibzadeh SZ, Salehzadeh A, Moradi-Shoeili Z, Shandiz SAS (2022) Iron oxide nanoparticles functionalized with 3-chloropropyltrimethoxysilane and conjugated with thiazole alter the expression of BAX, BCL2, and p53 genes in AGS cell line. Inorg Nano-Metal Chem. https://doi.org/10.1080/24701556.2021.2025074
Hosseinkhah M, Ghasemian R, Shokrollahi F, Mojdehi SR, Noveiri MJS, Hedayati M, Salehzadeh A (2021) Cytotoxic potential of nickel oxide nanoparticles functionalized with glutamic acid and conjugated with thiosemicarbazide (NiO@ Glu/TSC) against human gastric cancer cells. J Clust Sci. https://doi.org/10.1007/s10876-021-02124-2
Kachur K, Suntres Z (2020) The antibacterial properties of phenolic isomers, carvacrol and thymol. Crit Rev Food Sci Nutr 60(18):3042–3053. https://doi.org/10.1080/10408398.2019.1675585
Kazemi-Pasarvi S, Ebrahimi NG, Shahrampour D, Arab-Bafrani Z (2020) Reducing cytotoxicity of poly (lactic acid)-based/zinc oxide nanocomposites while boosting their antibacterial activities by thymol for biomedical applications. Int J Biol Macromol 164:4556–4565. https://doi.org/10.1016/j.ijbiomac.2020.09.069
Kumari R, Saini AK, Kumar A, Saini RV (2020) Apoptosis induction in lung and prostate cancer cells through silver nanoparticles synthesized from Pinus roxburghii bioactive fraction. J Biol Inorg Chem 25(1):23–37. https://doi.org/10.1007/s00775-019-01729-3
Marchese A, Orhan IE, Daglia M, Barbieri R, Di Lorenzo A, Nabavi SF, Nabavi SM (2016) Antibacterial and antifungal activities of thymol: a brief review of the literature. Food Chem 210:402–414. https://doi.org/10.1016/j.foodchem.2016.04.111
Mazur P, Skiba-Kurek I, Mrowiec P, Karczewska E, Drożdż R (2020) Synergistic ROS-associated antimicrobial activity of silver nanoparticles and gentamicin against Staphylococcus epidermidis. Int J Nanomed 15:3551–3562
Najafloo R, Behyari M, Imani R, Nour S (2020) A mini-review of thymol incorporated materials: applications in antibacterial wound dressing. J Drug Delivery Sci Technol 60:101904. https://doi.org/10.1016/j.jddst.2020.101904
Nejabatdoust A, Zamani H, Salehzadeh A (2019) Functionalization of ZnO nanoparticles by glutamic acid and conjugation with thiosemicarbazide alters expression of efflux pump genes in multiple drug-resistant Staphylococcus aureus strains. Microb Drug Resist 25(7):966–974. https://doi.org/10.1089/mdr.2018.0304
Perlman H, Zhang X, Chen MW, Walsh K, Buttyan R (1999) An elevated bax/bcl-2 ratio corresponds with the onset of prostate epithelial cell apoptosis. Cell Death Differ 6(1):48–54. https://doi.org/10.1038/sj.cdd.4400453
Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res 29:e45. https://doi.org/10.1093/nar/29.9.e45
Sangaiya P, Jayaprakash R (2018) A review on iron oxide nanoparticles and their biomedical applications. J Supercond Novel Magn 31(11):3397–3413. https://doi.org/10.1007/s10948-018-4841-2
Seraj FQM, Heravi-Faz N, Soltani A, Ahmadi SS, Talebpour A, Afshari AR, Bahrami A (2022) Thymol has anticancer effects in U-87 human malignant glioblastoma cells. Mol Biol Rep. https://doi.org/10.1007/s11033-022-07867-3
Seresht HR, Albadry BJ, Al-mosawi AKM, Gholami O, Cheshomi H (2019) The cytotoxic effects of thymol as the major component of trachyspermum ammi on breast cancer (MCF-7) cells. Pharm Chem J 53(2):101–107. https://doi.org/10.1007/s11094-019-01961-w
Shahriarinour M, Divsar F, Eskandari Z (2019) Synthesis, characterization, and antibacterial activity of thymol loaded SBA-15 mesoporous silica nanoparticles. Inorg Nano-Metal Chem 49(6):182–189. https://doi.org/10.1080/24701556.2019.1624569
Shin J, Naskar A, Ko D, Kim S, Kim KS (2022) Bioconjugated thymol-zinc oxide nanocomposite as a selective and biocompatible antibacterial agent against Staphylococcus Species. Int J Mol Sci 23(12):6770. https://doi.org/10.3390/ijms23126770
Someda M, Kuroki S, Miyachi H, Tachibana M, Yonehara S (2020) Caspase-8, receptor-interacting protein kinase 1 (RIPK1), and RIPK3 regulate retinoic acid-induced cell differentiation and necroptosis. Cell Death Differ 27(5):1539–1553. https://doi.org/10.1038/s41418-019-0434-2
Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: Cancer J Clin 71(3):209–249. https://doi.org/10.3322/caac.21660
Tian L, Wang X, Liu R, Zhang D, Wang X, Sun R, Gong G (2021) Antibacterial mechanism of thymol against Enterobacter sakazakii. Food Control 123:107716. https://doi.org/10.1016/j.foodcont.2020.107716
Vangijzegem T, Stanicki D, Laurent S (2019) Magnetic iron oxide nanoparticles for drug delivery: applications and characteristics. Expert Opin Drug Deliv 16(1):69–78. https://doi.org/10.1080/17425247.2019.1554647
Xu Y, Liu L, Qiu X, Liu Z, Li H, Li Z, Wang E (2012) CCL21/CCR7 prevents apoptosis via the ERK pathway in human non-small cell lung cancer cells. PLoS ONE 7(3):e33262. https://doi.org/10.1371/journal.pone.0033262
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AS and SHFK: Conceptualization. AS: Methodology. AS and HZ: Formal analysis and investigation. AS, SHFK and HZ: Writing Original Draft Preparation. AS, SHFK and HZ: Editing. AS: Resources. AS and HZ: Supervision.
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Fekri Kohan, S., Zamani, H. & Salehzadeh, A. Antibacterial potential and cytotoxic activity of iron oxide nanoparticles conjugated with thymol (Fe3O4@Glu-Thymol) on breast cancer cells and investigating the expression of BAX, CASP8, and BCL-2 genes. Biometals 36, 1273–1284 (2023). https://doi.org/10.1007/s10534-023-00516-7
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DOI: https://doi.org/10.1007/s10534-023-00516-7